US20220169976A1 - Probiotic bifidobacterium strain - Google Patents
Probiotic bifidobacterium strain Download PDFInfo
- Publication number
- US20220169976A1 US20220169976A1 US17/545,148 US202117545148A US2022169976A1 US 20220169976 A1 US20220169976 A1 US 20220169976A1 US 202117545148 A US202117545148 A US 202117545148A US 2022169976 A1 US2022169976 A1 US 2022169976A1
- Authority
- US
- United States
- Prior art keywords
- formulation
- subject
- strain
- inflammatory
- bifidobacterium
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Abandoned
Links
- 241000186000 Bifidobacterium Species 0.000 title abstract description 66
- 239000006041 probiotic Substances 0.000 title abstract description 29
- 235000018291 probiotics Nutrition 0.000 title abstract description 29
- 230000000529 probiotic effect Effects 0.000 title abstract description 27
- 239000000203 mixture Substances 0.000 claims description 65
- 238000009472 formulation Methods 0.000 claims description 61
- 102000004127 Cytokines Human genes 0.000 claims description 59
- 108090000695 Cytokines Proteins 0.000 claims description 59
- 238000000034 method Methods 0.000 claims description 36
- 241001608472 Bifidobacterium longum Species 0.000 claims description 34
- 102000013462 Interleukin-12 Human genes 0.000 claims description 29
- 108010065805 Interleukin-12 Proteins 0.000 claims description 29
- 102000003814 Interleukin-10 Human genes 0.000 claims description 28
- 108090000174 Interleukin-10 Proteins 0.000 claims description 28
- 229940009291 bifidobacterium longum Drugs 0.000 claims description 23
- 102000000852 Tumor Necrosis Factor-alpha Human genes 0.000 claims description 19
- 241000282414 Homo sapiens Species 0.000 claims description 18
- 206010061218 Inflammation Diseases 0.000 claims description 18
- 108060008682 Tumor Necrosis Factor Proteins 0.000 claims description 18
- 230000004054 inflammatory process Effects 0.000 claims description 18
- 230000003110 anti-inflammatory effect Effects 0.000 claims description 16
- 238000004519 manufacturing process Methods 0.000 claims description 14
- 108010074328 Interferon-gamma Proteins 0.000 claims description 12
- 102100037850 Interferon gamma Human genes 0.000 claims description 10
- MZOFCQQQCNRIBI-VMXHOPILSA-N (3s)-4-[[(2s)-1-[[(2s)-1-[[(1s)-1-carboxy-2-hydroxyethyl]amino]-4-methyl-1-oxopentan-2-yl]amino]-5-(diaminomethylideneamino)-1-oxopentan-2-yl]amino]-3-[[2-[[(2s)-2,6-diaminohexanoyl]amino]acetyl]amino]-4-oxobutanoic acid Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CC(O)=O)NC(=O)CNC(=O)[C@@H](N)CCCCN MZOFCQQQCNRIBI-VMXHOPILSA-N 0.000 claims description 9
- 239000000843 powder Substances 0.000 claims description 9
- 235000013361 beverage Nutrition 0.000 claims description 5
- 235000013305 food Nutrition 0.000 claims description 5
- 210000005095 gastrointestinal system Anatomy 0.000 claims description 5
- 239000002775 capsule Substances 0.000 claims description 4
- 235000013365 dairy product Nutrition 0.000 claims 1
- 239000003795 chemical substances by application Substances 0.000 abstract description 13
- 230000002519 immonomodulatory effect Effects 0.000 abstract description 12
- 241000699670 Mus sp. Species 0.000 description 44
- 241000894006 Bacteria Species 0.000 description 42
- 210000004027 cell Anatomy 0.000 description 40
- 238000011282 treatment Methods 0.000 description 37
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 35
- 238000011321 prophylaxis Methods 0.000 description 26
- 230000002757 inflammatory effect Effects 0.000 description 24
- 210000001035 gastrointestinal tract Anatomy 0.000 description 23
- 210000003819 peripheral blood mononuclear cell Anatomy 0.000 description 22
- 201000010099 disease Diseases 0.000 description 21
- 230000000694 effects Effects 0.000 description 21
- 229940076144 interleukin-10 Drugs 0.000 description 21
- 230000001580 bacterial effect Effects 0.000 description 19
- 241001465754 Metazoa Species 0.000 description 18
- 229940117681 interleukin-12 Drugs 0.000 description 18
- 238000001727 in vivo Methods 0.000 description 17
- 230000012010 growth Effects 0.000 description 16
- 206010028980 Neoplasm Diseases 0.000 description 15
- 108091023242 Internal transcribed spacer Proteins 0.000 description 14
- 208000035475 disorder Diseases 0.000 description 14
- 230000000770 proinflammatory effect Effects 0.000 description 14
- JQXXHWHPUNPDRT-WLSIYKJHSA-N rifampicin Chemical compound O([C@](C1=O)(C)O/C=C/[C@@H]([C@H]([C@@H](OC(C)=O)[C@H](C)[C@H](O)[C@H](C)[C@@H](O)[C@@H](C)\C=C\C=C(C)/C(=O)NC=2C(O)=C3C([O-])=C4C)C)OC)C4=C1C3=C(O)C=2\C=N\N1CC[NH+](C)CC1 JQXXHWHPUNPDRT-WLSIYKJHSA-N 0.000 description 14
- 229960001225 rifampicin Drugs 0.000 description 14
- 229920001817 Agar Polymers 0.000 description 13
- 239000008272 agar Substances 0.000 description 13
- LOKCTEFSRHRXRJ-UHFFFAOYSA-I dipotassium trisodium dihydrogen phosphate hydrogen phosphate dichloride Chemical compound P(=O)(O)(O)[O-].[K+].P(=O)(O)([O-])[O-].[Na+].[Na+].[Cl-].[K+].[Cl-].[Na+] LOKCTEFSRHRXRJ-UHFFFAOYSA-I 0.000 description 13
- 230000028709 inflammatory response Effects 0.000 description 13
- 239000002953 phosphate buffered saline Substances 0.000 description 13
- 238000012360 testing method Methods 0.000 description 13
- 210000001519 tissue Anatomy 0.000 description 13
- 206010012735 Diarrhoea Diseases 0.000 description 12
- 208000008589 Obesity Diseases 0.000 description 12
- 235000020824 obesity Nutrition 0.000 description 12
- 208000022559 Inflammatory bowel disease Diseases 0.000 description 11
- 238000000338 in vitro Methods 0.000 description 11
- 244000005700 microbiome Species 0.000 description 11
- 108090000623 proteins and genes Proteins 0.000 description 11
- 208000019901 Anxiety disease Diseases 0.000 description 10
- 238000003556 assay Methods 0.000 description 10
- 238000002360 preparation method Methods 0.000 description 10
- 210000004988 splenocyte Anatomy 0.000 description 10
- 102000004889 Interleukin-6 Human genes 0.000 description 9
- 230000003115 biocidal effect Effects 0.000 description 9
- 230000008033 biological extinction Effects 0.000 description 9
- 201000011510 cancer Diseases 0.000 description 9
- 230000001900 immune effect Effects 0.000 description 9
- 230000006698 induction Effects 0.000 description 9
- 239000000463 material Substances 0.000 description 9
- 230000015654 memory Effects 0.000 description 9
- 108090001005 Interleukin-6 Proteins 0.000 description 8
- 206010040047 Sepsis Diseases 0.000 description 8
- 230000028993 immune response Effects 0.000 description 8
- 210000000987 immune system Anatomy 0.000 description 8
- 235000013406 prebiotics Nutrition 0.000 description 8
- UCSJYZPVAKXKNQ-HZYVHMACSA-N streptomycin Chemical compound CN[C@H]1[C@H](O)[C@@H](O)[C@H](CO)O[C@H]1O[C@@H]1[C@](C=O)(O)[C@H](C)O[C@H]1O[C@@H]1[C@@H](NC(N)=N)[C@H](O)[C@@H](NC(N)=N)[C@H](O)[C@H]1O UCSJYZPVAKXKNQ-HZYVHMACSA-N 0.000 description 8
- 230000009885 systemic effect Effects 0.000 description 8
- 239000000427 antigen Substances 0.000 description 7
- 102000036639 antigens Human genes 0.000 description 7
- 108091007433 antigens Proteins 0.000 description 7
- 230000036506 anxiety Effects 0.000 description 7
- 150000001875 compounds Chemical class 0.000 description 7
- 239000003814 drug Substances 0.000 description 7
- 230000008014 freezing Effects 0.000 description 7
- 238000007710 freezing Methods 0.000 description 7
- 230000002496 gastric effect Effects 0.000 description 7
- 238000011534 incubation Methods 0.000 description 7
- 208000027866 inflammatory disease Diseases 0.000 description 7
- 238000010172 mouse model Methods 0.000 description 7
- 239000000902 placebo Substances 0.000 description 7
- 229940068196 placebo Drugs 0.000 description 7
- 210000002966 serum Anatomy 0.000 description 7
- 108020004414 DNA Proteins 0.000 description 6
- 101000914514 Homo sapiens T-cell-specific surface glycoprotein CD28 Proteins 0.000 description 6
- 108091028043 Nucleic acid sequence Proteins 0.000 description 6
- 102100027213 T-cell-specific surface glycoprotein CD28 Human genes 0.000 description 6
- 239000000935 antidepressant agent Substances 0.000 description 6
- 229940005513 antidepressants Drugs 0.000 description 6
- 238000011161 development Methods 0.000 description 6
- 230000018109 developmental process Effects 0.000 description 6
- 230000000968 intestinal effect Effects 0.000 description 6
- 230000007246 mechanism Effects 0.000 description 6
- 244000052769 pathogen Species 0.000 description 6
- 239000000047 product Substances 0.000 description 6
- 235000018102 proteins Nutrition 0.000 description 6
- 102000004169 proteins and genes Human genes 0.000 description 6
- 206010039073 rheumatoid arthritis Diseases 0.000 description 6
- 210000000952 spleen Anatomy 0.000 description 6
- 208000023275 Autoimmune disease Diseases 0.000 description 5
- XUJNEKJLAYXESH-REOHCLBHSA-N L-Cysteine Chemical compound SC[C@H](N)C(O)=O XUJNEKJLAYXESH-REOHCLBHSA-N 0.000 description 5
- 241001529936 Murinae Species 0.000 description 5
- JGSARLDLIJGVTE-MBNYWOFBSA-N Penicillin G Chemical compound N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)CC1=CC=CC=C1 JGSARLDLIJGVTE-MBNYWOFBSA-N 0.000 description 5
- 230000000845 anti-microbial effect Effects 0.000 description 5
- 230000008901 benefit Effects 0.000 description 5
- 210000000941 bile Anatomy 0.000 description 5
- 238000006243 chemical reaction Methods 0.000 description 5
- 206010009887 colitis Diseases 0.000 description 5
- 239000000306 component Substances 0.000 description 5
- 230000003750 conditioning effect Effects 0.000 description 5
- 229960002433 cysteine Drugs 0.000 description 5
- 208000015181 infectious disease Diseases 0.000 description 5
- 208000002551 irritable bowel syndrome Diseases 0.000 description 5
- 230000001717 pathogenic effect Effects 0.000 description 5
- 230000004044 response Effects 0.000 description 5
- 239000006228 supernatant Substances 0.000 description 5
- 239000000725 suspension Substances 0.000 description 5
- 208000001072 type 2 diabetes mellitus Diseases 0.000 description 5
- 239000003981 vehicle Substances 0.000 description 5
- 208000024827 Alzheimer disease Diseases 0.000 description 4
- 241000186015 Bifidobacterium longum subsp. infantis Species 0.000 description 4
- 206010020751 Hypersensitivity Diseases 0.000 description 4
- 208000021384 Obsessive-Compulsive disease Diseases 0.000 description 4
- 230000007815 allergy Effects 0.000 description 4
- 230000004075 alteration Effects 0.000 description 4
- 238000004458 analytical method Methods 0.000 description 4
- 208000006673 asthma Diseases 0.000 description 4
- 230000004888 barrier function Effects 0.000 description 4
- 210000001072 colon Anatomy 0.000 description 4
- 230000007423 decrease Effects 0.000 description 4
- 230000003247 decreasing effect Effects 0.000 description 4
- 235000005911 diet Nutrition 0.000 description 4
- 229940079593 drug Drugs 0.000 description 4
- 238000002474 experimental method Methods 0.000 description 4
- ZDXPYRJPNDTMRX-UHFFFAOYSA-N glutamine Natural products OC(=O)C(N)CCC(N)=O ZDXPYRJPNDTMRX-UHFFFAOYSA-N 0.000 description 4
- 239000012678 infectious agent Substances 0.000 description 4
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 4
- 210000002540 macrophage Anatomy 0.000 description 4
- 235000013336 milk Nutrition 0.000 description 4
- 239000008267 milk Substances 0.000 description 4
- 210000004080 milk Anatomy 0.000 description 4
- 108090000765 processed proteins & peptides Proteins 0.000 description 4
- 239000000243 solution Substances 0.000 description 4
- 230000002269 spontaneous effect Effects 0.000 description 4
- 229960005322 streptomycin Drugs 0.000 description 4
- 229960005486 vaccine Drugs 0.000 description 4
- 235000013618 yogurt Nutrition 0.000 description 4
- 206010006895 Cachexia Diseases 0.000 description 3
- 102000016938 Catalase Human genes 0.000 description 3
- 108010053835 Catalase Proteins 0.000 description 3
- 241000193163 Clostridioides difficile Species 0.000 description 3
- 206010009900 Colitis ulcerative Diseases 0.000 description 3
- 241000588724 Escherichia coli Species 0.000 description 3
- 230000005526 G1 to G0 transition Effects 0.000 description 3
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 3
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 3
- 241000282412 Homo Species 0.000 description 3
- 206010022489 Insulin Resistance Diseases 0.000 description 3
- 229930182555 Penicillin Natural products 0.000 description 3
- 201000006704 Ulcerative Colitis Diseases 0.000 description 3
- 101000872823 Xenopus laevis Probable histone deacetylase 1-A Proteins 0.000 description 3
- 230000001464 adherent effect Effects 0.000 description 3
- 239000002671 adjuvant Substances 0.000 description 3
- 208000026935 allergic disease Diseases 0.000 description 3
- 238000010171 animal model Methods 0.000 description 3
- 239000003242 anti bacterial agent Substances 0.000 description 3
- 230000001430 anti-depressive effect Effects 0.000 description 3
- 229940088710 antibiotic agent Drugs 0.000 description 3
- 238000003149 assay kit Methods 0.000 description 3
- 230000006399 behavior Effects 0.000 description 3
- 239000003782 beta lactam antibiotic agent Substances 0.000 description 3
- 238000001574 biopsy Methods 0.000 description 3
- 210000004369 blood Anatomy 0.000 description 3
- 239000008280 blood Substances 0.000 description 3
- 210000003169 central nervous system Anatomy 0.000 description 3
- 230000000112 colonic effect Effects 0.000 description 3
- IQFVPQOLBLOTPF-HKXUKFGYSA-L congo red Chemical compound [Na+].[Na+].C1=CC=CC2=C(N)C(/N=N/C3=CC=C(C=C3)C3=CC=C(C=C3)/N=N/C3=C(C4=CC=CC=C4C(=C3)S([O-])(=O)=O)N)=CC(S([O-])(=O)=O)=C21 IQFVPQOLBLOTPF-HKXUKFGYSA-L 0.000 description 3
- 239000012228 culture supernatant Substances 0.000 description 3
- 235000018417 cysteine Nutrition 0.000 description 3
- XUJNEKJLAYXESH-UHFFFAOYSA-N cysteine Natural products SCC(N)C(O)=O XUJNEKJLAYXESH-UHFFFAOYSA-N 0.000 description 3
- 230000006378 damage Effects 0.000 description 3
- 230000007123 defense Effects 0.000 description 3
- 230000001419 dependent effect Effects 0.000 description 3
- 206010012601 diabetes mellitus Diseases 0.000 description 3
- 230000037213 diet Effects 0.000 description 3
- 239000003937 drug carrier Substances 0.000 description 3
- 229960004341 escitalopram Drugs 0.000 description 3
- WSEQXVZVJXJVFP-FQEVSTJZSA-N escitalopram Chemical compound C1([C@]2(C3=CC=C(C=C3CO2)C#N)CCCN(C)C)=CC=C(F)C=C1 WSEQXVZVJXJVFP-FQEVSTJZSA-N 0.000 description 3
- 239000008103 glucose Substances 0.000 description 3
- 244000005709 gut microbiome Species 0.000 description 3
- 235000009200 high fat diet Nutrition 0.000 description 3
- 238000003384 imaging method Methods 0.000 description 3
- 238000003780 insertion Methods 0.000 description 3
- 230000037431 insertion Effects 0.000 description 3
- 230000006799 invasive growth in response to glucose limitation Effects 0.000 description 3
- 150000002632 lipids Chemical class 0.000 description 3
- 230000007774 longterm Effects 0.000 description 3
- 239000004579 marble Substances 0.000 description 3
- 208000008338 non-alcoholic fatty liver disease Diseases 0.000 description 3
- 239000002773 nucleotide Substances 0.000 description 3
- 230000007170 pathology Effects 0.000 description 3
- 229940049954 penicillin Drugs 0.000 description 3
- 230000008569 process Effects 0.000 description 3
- 229940124834 selective serotonin reuptake inhibitor Drugs 0.000 description 3
- 230000035945 sensitivity Effects 0.000 description 3
- 210000000813 small intestine Anatomy 0.000 description 3
- 241000894007 species Species 0.000 description 3
- 210000002784 stomach Anatomy 0.000 description 3
- 230000004083 survival effect Effects 0.000 description 3
- 239000003826 tablet Substances 0.000 description 3
- 108010044241 tetanus toxin fragment C Proteins 0.000 description 3
- 230000001225 therapeutic effect Effects 0.000 description 3
- 238000002560 therapeutic procedure Methods 0.000 description 3
- 238000002255 vaccination Methods 0.000 description 3
- MYPYJXKWCTUITO-LYRMYLQWSA-N vancomycin Chemical compound O([C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC1=C2C=C3C=C1OC1=CC=C(C=C1Cl)[C@@H](O)[C@H](C(N[C@@H](CC(N)=O)C(=O)N[C@H]3C(=O)N[C@H]1C(=O)N[C@H](C(N[C@@H](C3=CC(O)=CC(O)=C3C=3C(O)=CC=C1C=3)C(O)=O)=O)[C@H](O)C1=CC=C(C(=C1)Cl)O2)=O)NC(=O)[C@@H](CC(C)C)NC)[C@H]1C[C@](C)(N)[C@H](O)[C@H](C)O1 MYPYJXKWCTUITO-LYRMYLQWSA-N 0.000 description 3
- 239000013598 vector Substances 0.000 description 3
- 208000016261 weight loss Diseases 0.000 description 3
- 230000004580 weight loss Effects 0.000 description 3
- 239000002132 β-lactam antibiotic Substances 0.000 description 3
- 229940124586 β-lactam antibiotics Drugs 0.000 description 3
- MINDHVHHQZYEEK-UHFFFAOYSA-N (E)-(2S,3R,4R,5S)-5-[(2S,3S,4S,5S)-2,3-epoxy-5-hydroxy-4-methylhexyl]tetrahydro-3,4-dihydroxy-(beta)-methyl-2H-pyran-2-crotonic acid ester with 9-hydroxynonanoic acid Natural products CC(O)C(C)C1OC1CC1C(O)C(O)C(CC(C)=CC(=O)OCCCCCCCCC(O)=O)OC1 MINDHVHHQZYEEK-UHFFFAOYSA-N 0.000 description 2
- 208000002874 Acne Vulgaris Diseases 0.000 description 2
- HJCMDXDYPOUFDY-WHFBIAKZSA-N Ala-Gln Chemical compound C[C@H](N)C(=O)N[C@H](C(O)=O)CCC(N)=O HJCMDXDYPOUFDY-WHFBIAKZSA-N 0.000 description 2
- 206010003011 Appendicitis Diseases 0.000 description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 208000035143 Bacterial infection Diseases 0.000 description 2
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 2
- 238000009631 Broth culture Methods 0.000 description 2
- 241000589875 Campylobacter jejuni Species 0.000 description 2
- 241001112696 Clostridia Species 0.000 description 2
- 208000015943 Coeliac disease Diseases 0.000 description 2
- 208000011231 Crohn disease Diseases 0.000 description 2
- WQZGKKKJIJFFOK-QTVWNMPRSA-N D-mannopyranose Chemical compound OC[C@H]1OC(O)[C@@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-QTVWNMPRSA-N 0.000 description 2
- SRBFZHDQGSBBOR-IOVATXLUSA-N D-xylopyranose Chemical compound O[C@@H]1COC(O)[C@H](O)[C@H]1O SRBFZHDQGSBBOR-IOVATXLUSA-N 0.000 description 2
- 208000017701 Endocrine disease Diseases 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 2
- 108010052764 Fructose-6-phosphate phosphoketolase Proteins 0.000 description 2
- 206010017533 Fungal infection Diseases 0.000 description 2
- 206010017993 Gastrointestinal neoplasms Diseases 0.000 description 2
- 208000031886 HIV Infections Diseases 0.000 description 2
- 208000037357 HIV infectious disease Diseases 0.000 description 2
- 101001076408 Homo sapiens Interleukin-6 Proteins 0.000 description 2
- 101000611183 Homo sapiens Tumor necrosis factor Proteins 0.000 description 2
- 101150101999 IL6 gene Proteins 0.000 description 2
- 206010061598 Immunodeficiency Diseases 0.000 description 2
- 208000029462 Immunodeficiency disease Diseases 0.000 description 2
- 102000008070 Interferon-gamma Human genes 0.000 description 2
- 102000004890 Interleukin-8 Human genes 0.000 description 2
- 108090001007 Interleukin-8 Proteins 0.000 description 2
- WHUUTDBJXJRKMK-VKHMYHEASA-N L-glutamic acid Chemical compound OC(=O)[C@@H](N)CCC(O)=O WHUUTDBJXJRKMK-VKHMYHEASA-N 0.000 description 2
- 241000186660 Lactobacillus Species 0.000 description 2
- 241000194035 Lactococcus lactis Species 0.000 description 2
- 208000031888 Mycoses Diseases 0.000 description 2
- 208000001132 Osteoporosis Diseases 0.000 description 2
- 208000002389 Pouchitis Diseases 0.000 description 2
- 201000004681 Psoriasis Diseases 0.000 description 2
- 206010037660 Pyrexia Diseases 0.000 description 2
- LCTONWCANYUPML-UHFFFAOYSA-M Pyruvate Chemical compound CC(=O)C([O-])=O LCTONWCANYUPML-UHFFFAOYSA-M 0.000 description 2
- 241000702670 Rotavirus Species 0.000 description 2
- 241000293869 Salmonella enterica subsp. enterica serovar Typhimurium Species 0.000 description 2
- 208000019802 Sexually transmitted disease Diseases 0.000 description 2
- 241000194026 Streptococcus gordonii Species 0.000 description 2
- 229940123317 Sulfonamide antibiotic Drugs 0.000 description 2
- 208000025865 Ulcer Diseases 0.000 description 2
- 208000025609 Urogenital disease Diseases 0.000 description 2
- 108010059993 Vancomycin Proteins 0.000 description 2
- 208000036142 Viral infection Diseases 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 235000020167 acidified milk Nutrition 0.000 description 2
- 206010000496 acne Diseases 0.000 description 2
- 230000009471 action Effects 0.000 description 2
- 239000004480 active ingredient Substances 0.000 description 2
- 229940126574 aminoglycoside antibiotic Drugs 0.000 description 2
- 239000002647 aminoglycoside antibiotic agent Substances 0.000 description 2
- 208000007502 anemia Diseases 0.000 description 2
- 208000022531 anorexia Diseases 0.000 description 2
- 230000003042 antagnostic effect Effects 0.000 description 2
- 230000002924 anti-infective effect Effects 0.000 description 2
- 230000030741 antigen processing and presentation Effects 0.000 description 2
- 230000005784 autoimmunity Effects 0.000 description 2
- 208000022362 bacterial infectious disease Diseases 0.000 description 2
- 230000003542 behavioural effect Effects 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 239000003833 bile salt Substances 0.000 description 2
- 229940093761 bile salts Drugs 0.000 description 2
- 230000000975 bioactive effect Effects 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 230000023555 blood coagulation Effects 0.000 description 2
- 230000036760 body temperature Effects 0.000 description 2
- 150000001720 carbohydrates Chemical class 0.000 description 2
- 238000005119 centrifugation Methods 0.000 description 2
- 235000013351 cheese Nutrition 0.000 description 2
- 238000003501 co-culture Methods 0.000 description 2
- 239000012141 concentrate Substances 0.000 description 2
- 235000008504 concentrate Nutrition 0.000 description 2
- 230000001276 controlling effect Effects 0.000 description 2
- 208000029078 coronary artery disease Diseases 0.000 description 2
- 230000001351 cycling effect Effects 0.000 description 2
- 230000016396 cytokine production Effects 0.000 description 2
- 206010061428 decreased appetite Diseases 0.000 description 2
- 230000008260 defense mechanism Effects 0.000 description 2
- 239000003651 drinking water Substances 0.000 description 2
- 235000020188 drinking water Nutrition 0.000 description 2
- 230000007613 environmental effect Effects 0.000 description 2
- 229940088598 enzyme Drugs 0.000 description 2
- 239000012091 fetal bovine serum Substances 0.000 description 2
- 230000037406 food intake Effects 0.000 description 2
- 230000006870 function Effects 0.000 description 2
- 230000004077 genetic alteration Effects 0.000 description 2
- 231100000118 genetic alteration Toxicity 0.000 description 2
- 229930195712 glutamate Natural products 0.000 description 2
- 229940088597 hormone Drugs 0.000 description 2
- 239000005556 hormone Substances 0.000 description 2
- 208000033519 human immunodeficiency virus infectious disease Diseases 0.000 description 2
- JYGXADMDTFJGBT-VWUMJDOOSA-N hydrocortisone Chemical compound O=C1CC[C@]2(C)[C@H]3[C@@H](O)C[C@](C)([C@@](CC4)(O)C(=O)CO)[C@@H]4[C@@H]3CCC2=C1 JYGXADMDTFJGBT-VWUMJDOOSA-N 0.000 description 2
- 238000002649 immunization Methods 0.000 description 2
- 230000007813 immunodeficiency Effects 0.000 description 2
- 230000016784 immunoglobulin production Effects 0.000 description 2
- 230000004957 immunoregulator effect Effects 0.000 description 2
- 230000001524 infective effect Effects 0.000 description 2
- 229960000598 infliximab Drugs 0.000 description 2
- 208000014674 injury Diseases 0.000 description 2
- 229910052500 inorganic mineral Inorganic materials 0.000 description 2
- 229960003130 interferon gamma Drugs 0.000 description 2
- 229940100601 interleukin-6 Drugs 0.000 description 2
- XKTZWUACRZHVAN-VADRZIEHSA-N interleukin-8 Chemical compound C([C@H](NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC=1C2=CC=CC=C2NC=1)NC(=O)[C@@H](NC(C)=O)CCSC)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H]([C@@H](C)O)C(=O)NCC(=O)N[C@@H](CCSC)C(=O)N1[C@H](CCC1)C(=O)N1[C@H](CCC1)C(=O)N[C@@H](C)C(=O)N[C@H](CC(O)=O)C(=O)N[C@H](CCC(O)=O)C(=O)N[C@H](CC(O)=O)C(=O)N[C@H](CC=1C=CC(O)=CC=1)C(=O)N[C@H](CO)C(=O)N1[C@H](CCC1)C(N)=O)C1=CC=CC=C1 XKTZWUACRZHVAN-VADRZIEHSA-N 0.000 description 2
- 239000007928 intraperitoneal injection Substances 0.000 description 2
- 208000028867 ischemia Diseases 0.000 description 2
- 238000002955 isolation Methods 0.000 description 2
- 208000017169 kidney disease Diseases 0.000 description 2
- 235000014655 lactic acid Nutrition 0.000 description 2
- 239000004310 lactic acid Substances 0.000 description 2
- 210000000265 leukocyte Anatomy 0.000 description 2
- 150000002617 leukotrienes Chemical class 0.000 description 2
- 208000019423 liver disease Diseases 0.000 description 2
- 210000004698 lymphocyte Anatomy 0.000 description 2
- 239000003550 marker Substances 0.000 description 2
- 230000010120 metabolic dysregulation Effects 0.000 description 2
- 206010061289 metastatic neoplasm Diseases 0.000 description 2
- 230000000813 microbial effect Effects 0.000 description 2
- 235000010755 mineral Nutrition 0.000 description 2
- 239000002366 mineral element Substances 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 210000001616 monocyte Anatomy 0.000 description 2
- 201000006417 multiple sclerosis Diseases 0.000 description 2
- 229960003128 mupirocin Drugs 0.000 description 2
- 229930187697 mupirocin Natural products 0.000 description 2
- DDHVILIIHBIMQU-YJGQQKNPSA-L mupirocin calcium hydrate Chemical compound O.O.[Ca+2].C[C@H](O)[C@H](C)[C@@H]1O[C@H]1C[C@@H]1[C@@H](O)[C@@H](O)[C@H](C\C(C)=C\C(=O)OCCCCCCCCC([O-])=O)OC1.C[C@H](O)[C@H](C)[C@@H]1O[C@H]1C[C@@H]1[C@@H](O)[C@@H](O)[C@H](C\C(C)=C\C(=O)OCCCCCCCCC([O-])=O)OC1 DDHVILIIHBIMQU-YJGQQKNPSA-L 0.000 description 2
- 229940021182 non-steroidal anti-inflammatory drug Drugs 0.000 description 2
- 125000003729 nucleotide group Chemical group 0.000 description 2
- 208000030212 nutrition disease Diseases 0.000 description 2
- 210000000056 organ Anatomy 0.000 description 2
- 229940094443 oxytocics prostaglandins Drugs 0.000 description 2
- 208000019906 panic disease Diseases 0.000 description 2
- 208000028169 periodontal disease Diseases 0.000 description 2
- 210000005259 peripheral blood Anatomy 0.000 description 2
- 239000011886 peripheral blood Substances 0.000 description 2
- 208000028173 post-traumatic stress disease Diseases 0.000 description 2
- 230000002265 prevention Effects 0.000 description 2
- 102000004196 processed proteins & peptides Human genes 0.000 description 2
- 150000003180 prostaglandins Chemical class 0.000 description 2
- 230000001681 protective effect Effects 0.000 description 2
- 210000003289 regulatory T cell Anatomy 0.000 description 2
- 230000001105 regulatory effect Effects 0.000 description 2
- 230000010076 replication Effects 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 208000023504 respiratory system disease Diseases 0.000 description 2
- 230000004043 responsiveness Effects 0.000 description 2
- 230000002441 reversible effect Effects 0.000 description 2
- 239000012896 selective serotonin reuptake inhibitor Substances 0.000 description 2
- 238000012163 sequencing technique Methods 0.000 description 2
- 230000000638 stimulation Effects 0.000 description 2
- 229960005404 sulfamethoxazole Drugs 0.000 description 2
- JLKIGFTWXXRPMT-UHFFFAOYSA-N sulphamethoxazole Chemical compound O1C(C)=CC(NS(=O)(=O)C=2C=CC(N)=CC=2)=N1 JLKIGFTWXXRPMT-UHFFFAOYSA-N 0.000 description 2
- 208000024891 symptom Diseases 0.000 description 2
- 239000011573 trace mineral Substances 0.000 description 2
- 235000013619 trace mineral Nutrition 0.000 description 2
- 238000002054 transplantation Methods 0.000 description 2
- 230000008733 trauma Effects 0.000 description 2
- IEDVJHCEMCRBQM-UHFFFAOYSA-N trimethoprim Chemical compound COC1=C(OC)C(OC)=CC(CC=2C(=NC(N)=NC=2)N)=C1 IEDVJHCEMCRBQM-UHFFFAOYSA-N 0.000 description 2
- 229960001082 trimethoprim Drugs 0.000 description 2
- 231100000397 ulcer Toxicity 0.000 description 2
- 229960003165 vancomycin Drugs 0.000 description 2
- MYPYJXKWCTUITO-UHFFFAOYSA-N vancomycin Natural products O1C(C(=C2)Cl)=CC=C2C(O)C(C(NC(C2=CC(O)=CC(O)=C2C=2C(O)=CC=C3C=2)C(O)=O)=O)NC(=O)C3NC(=O)C2NC(=O)C(CC(N)=O)NC(=O)C(NC(=O)C(CC(C)C)NC)C(O)C(C=C3Cl)=CC=C3OC3=CC2=CC1=C3OC1OC(CO)C(O)C(O)C1OC1CC(C)(N)C(O)C(C)O1 MYPYJXKWCTUITO-UHFFFAOYSA-N 0.000 description 2
- 230000009385 viral infection Effects 0.000 description 2
- 235000013343 vitamin Nutrition 0.000 description 2
- 239000011782 vitamin Substances 0.000 description 2
- 229940088594 vitamin Drugs 0.000 description 2
- 229930003231 vitamin Natural products 0.000 description 2
- 150000003722 vitamin derivatives Chemical class 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 230000029663 wound healing Effects 0.000 description 2
- KTGRHKOEFSJQNS-BDQAORGHSA-N (1s)-1-[3-(dimethylamino)propyl]-1-(4-fluorophenyl)-3h-2-benzofuran-5-carbonitrile;oxalic acid Chemical compound OC(=O)C(O)=O.C1([C@]2(C3=CC=C(C=C3CO2)C#N)CCCN(C)C)=CC=C(F)C=C1 KTGRHKOEFSJQNS-BDQAORGHSA-N 0.000 description 1
- LOGFVTREOLYCPF-KXNHARMFSA-N (2s,3r)-2-[[(2r)-1-[(2s)-2,6-diaminohexanoyl]pyrrolidine-2-carbonyl]amino]-3-hydroxybutanoic acid Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)[C@H]1CCCN1C(=O)[C@@H](N)CCCCN LOGFVTREOLYCPF-KXNHARMFSA-N 0.000 description 1
- RTHCYVBBDHJXIQ-MRXNPFEDSA-N (R)-fluoxetine Chemical compound O([C@H](CCNC)C=1C=CC=CC=1)C1=CC=C(C(F)(F)F)C=C1 RTHCYVBBDHJXIQ-MRXNPFEDSA-N 0.000 description 1
- YZEUHQHUFTYLPH-UHFFFAOYSA-N 2-nitroimidazole Chemical compound [O-][N+](=O)C1=NC=CN1 YZEUHQHUFTYLPH-UHFFFAOYSA-N 0.000 description 1
- QWZHDKGQKYEBKK-UHFFFAOYSA-N 3-aminochromen-2-one Chemical compound C1=CC=C2OC(=O)C(N)=CC2=C1 QWZHDKGQKYEBKK-UHFFFAOYSA-N 0.000 description 1
- 229920000936 Agarose Polymers 0.000 description 1
- 201000001320 Atherosclerosis Diseases 0.000 description 1
- 241000606125 Bacteroides Species 0.000 description 1
- 102000004506 Blood Proteins Human genes 0.000 description 1
- 108010017384 Blood Proteins Proteins 0.000 description 1
- 241000589562 Brucella Species 0.000 description 1
- 238000011752 CBA/J (JAX™ mouse strain) Methods 0.000 description 1
- 208000024172 Cardiovascular disease Diseases 0.000 description 1
- 102000019034 Chemokines Human genes 0.000 description 1
- 108010012236 Chemokines Proteins 0.000 description 1
- 241000193403 Clostridium Species 0.000 description 1
- OMFXVFTZEKFJBZ-UHFFFAOYSA-N Corticosterone Natural products O=C1CCC2(C)C3C(O)CC(C)(C(CC4)C(=O)CO)C4C3CCC2=C1 OMFXVFTZEKFJBZ-UHFFFAOYSA-N 0.000 description 1
- IGXWBGJHJZYPQS-SSDOTTSWSA-N D-Luciferin Chemical compound OC(=O)[C@H]1CSC(C=2SC3=CC=C(O)C=C3N=2)=N1 IGXWBGJHJZYPQS-SSDOTTSWSA-N 0.000 description 1
- 230000008836 DNA modification Effects 0.000 description 1
- 230000009946 DNA mutation Effects 0.000 description 1
- CYCGRDQQIOGCKX-UHFFFAOYSA-N Dehydro-luciferin Natural products OC(=O)C1=CSC(C=2SC3=CC(O)=CC=C3N=2)=N1 CYCGRDQQIOGCKX-UHFFFAOYSA-N 0.000 description 1
- 241001646719 Escherichia coli O157:H7 Species 0.000 description 1
- 102000010834 Extracellular Matrix Proteins Human genes 0.000 description 1
- 108010037362 Extracellular Matrix Proteins Proteins 0.000 description 1
- BJGNCJDXODQBOB-UHFFFAOYSA-N Fivefly Luciferin Natural products OC(=O)C1CSC(C=2SC3=CC(O)=CC=C3N=2)=N1 BJGNCJDXODQBOB-UHFFFAOYSA-N 0.000 description 1
- 229930091371 Fructose Natural products 0.000 description 1
- RFSUNEUAIZKAJO-ARQDHWQXSA-N Fructose Chemical compound OC[C@H]1O[C@](O)(CO)[C@@H](O)[C@@H]1O RFSUNEUAIZKAJO-ARQDHWQXSA-N 0.000 description 1
- 239000005715 Fructose Substances 0.000 description 1
- 208000011688 Generalised anxiety disease Diseases 0.000 description 1
- 229930182566 Gentamicin Natural products 0.000 description 1
- CEAZRRDELHUEMR-URQXQFDESA-N Gentamicin Chemical compound O1[C@H](C(C)NC)CC[C@@H](N)[C@H]1O[C@H]1[C@H](O)[C@@H](O[C@@H]2[C@@H]([C@@H](NC)[C@@](C)(O)CO2)O)[C@H](N)C[C@@H]1N CEAZRRDELHUEMR-URQXQFDESA-N 0.000 description 1
- 244000068988 Glycine max Species 0.000 description 1
- 235000010469 Glycine max Nutrition 0.000 description 1
- 238000003794 Gram staining Methods 0.000 description 1
- 101000887490 Homo sapiens Guanine nucleotide-binding protein G(z) subunit alpha Proteins 0.000 description 1
- 101000599940 Homo sapiens Interferon gamma Proteins 0.000 description 1
- 101001033249 Homo sapiens Interleukin-1 beta Proteins 0.000 description 1
- 101001033233 Homo sapiens Interleukin-10 Proteins 0.000 description 1
- 101001002657 Homo sapiens Interleukin-2 Proteins 0.000 description 1
- 101001055222 Homo sapiens Interleukin-8 Proteins 0.000 description 1
- 101000979342 Homo sapiens Nuclear factor NF-kappa-B p105 subunit Proteins 0.000 description 1
- 101000669447 Homo sapiens Toll-like receptor 4 Proteins 0.000 description 1
- 208000035150 Hypercholesterolemia Diseases 0.000 description 1
- 206010020772 Hypertension Diseases 0.000 description 1
- 101000668058 Infectious salmon anemia virus (isolate Atlantic salmon/Norway/810/9/99) RNA-directed RNA polymerase catalytic subunit Proteins 0.000 description 1
- 102000006992 Interferon-alpha Human genes 0.000 description 1
- 108010047761 Interferon-alpha Proteins 0.000 description 1
- 108090000193 Interleukin-1 beta Proteins 0.000 description 1
- 102000003777 Interleukin-1 beta Human genes 0.000 description 1
- 230000004163 JAK-STAT signaling pathway Effects 0.000 description 1
- 244000199866 Lactobacillus casei Species 0.000 description 1
- 235000013958 Lactobacillus casei Nutrition 0.000 description 1
- 108010092277 Leptin Proteins 0.000 description 1
- 102000016267 Leptin Human genes 0.000 description 1
- DDWFXDSYGUXRAY-UHFFFAOYSA-N Luciferin Natural products CCc1c(C)c(CC2NC(=O)C(=C2C=C)C)[nH]c1Cc3[nH]c4C(=C5/NC(CC(=O)O)C(C)C5CC(=O)O)CC(=O)c4c3C DDWFXDSYGUXRAY-UHFFFAOYSA-N 0.000 description 1
- 102000043131 MHC class II family Human genes 0.000 description 1
- 108091054438 MHC class II family Proteins 0.000 description 1
- 208000002720 Malnutrition Diseases 0.000 description 1
- RJQXTJLFIWVMTO-TYNCELHUSA-N Methicillin Chemical compound COC1=CC=CC(OC)=C1C(=O)N[C@@H]1C(=O)N2[C@@H](C(O)=O)C(C)(C)S[C@@H]21 RJQXTJLFIWVMTO-TYNCELHUSA-N 0.000 description 1
- 208000019022 Mood disease Diseases 0.000 description 1
- 206010028116 Mucosal inflammation Diseases 0.000 description 1
- 102000016943 Muramidase Human genes 0.000 description 1
- 108010014251 Muramidase Proteins 0.000 description 1
- 241000699666 Mus <mouse, genus> Species 0.000 description 1
- 241000699660 Mus musculus Species 0.000 description 1
- 101001033265 Mus musculus Interleukin-10 Proteins 0.000 description 1
- 101000648740 Mus musculus Tumor necrosis factor Proteins 0.000 description 1
- 108010062010 N-Acetylmuramoyl-L-alanine Amidase Proteins 0.000 description 1
- 108010057466 NF-kappa B Proteins 0.000 description 1
- 102000003945 NF-kappa B Human genes 0.000 description 1
- YJQPYGGHQPGBLI-UHFFFAOYSA-N Novobiocin Natural products O1C(C)(C)C(OC)C(OC(N)=O)C(O)C1OC1=CC=C(C(O)=C(NC(=O)C=2C=C(CC=C(C)C)C(O)=CC=2)C(=O)O2)C2=C1C YJQPYGGHQPGBLI-UHFFFAOYSA-N 0.000 description 1
- 102100023050 Nuclear factor NF-kappa-B p105 subunit Human genes 0.000 description 1
- KYGZCKSPAKDVKC-UHFFFAOYSA-N Oxolinic acid Chemical compound C1=C2N(CC)C=C(C(O)=O)C(=O)C2=CC2=C1OCO2 KYGZCKSPAKDVKC-UHFFFAOYSA-N 0.000 description 1
- 238000010222 PCR analysis Methods 0.000 description 1
- 241001085205 Prenanthella exigua Species 0.000 description 1
- 229930189077 Rifamycin Natural products 0.000 description 1
- 241000283984 Rodentia Species 0.000 description 1
- 238000012300 Sequence Analysis Methods 0.000 description 1
- 235000014897 Streptococcus lactis Nutrition 0.000 description 1
- 230000005867 T cell response Effects 0.000 description 1
- 210000000662 T-lymphocyte subset Anatomy 0.000 description 1
- 239000004098 Tetracycline Substances 0.000 description 1
- 102100039360 Toll-like receptor 4 Human genes 0.000 description 1
- 108091023040 Transcription factor Proteins 0.000 description 1
- 102000040945 Transcription factor Human genes 0.000 description 1
- 229920004890 Triton X-100 Polymers 0.000 description 1
- 239000013504 Triton X-100 Substances 0.000 description 1
- 102100040247 Tumor necrosis factor Human genes 0.000 description 1
- 241000021375 Xenogenes Species 0.000 description 1
- 230000001594 aberrant effect Effects 0.000 description 1
- 230000005856 abnormality Effects 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 239000011149 active material Substances 0.000 description 1
- 230000004721 adaptive immunity Effects 0.000 description 1
- 239000000654 additive Substances 0.000 description 1
- 239000002390 adhesive tape Substances 0.000 description 1
- 210000000577 adipose tissue Anatomy 0.000 description 1
- 230000002411 adverse Effects 0.000 description 1
- 239000011543 agarose gel Substances 0.000 description 1
- WQZGKKKJIJFFOK-PHYPRBDBSA-N alpha-D-galactose Chemical compound OC[C@H]1O[C@H](O)[C@H](O)[C@@H](O)[C@H]1O WQZGKKKJIJFFOK-PHYPRBDBSA-N 0.000 description 1
- 230000001668 ameliorated effect Effects 0.000 description 1
- 230000037354 amino acid metabolism Effects 0.000 description 1
- 150000001413 amino acids Chemical group 0.000 description 1
- 229960000723 ampicillin Drugs 0.000 description 1
- AVKUERGKIZMTKX-NJBDSQKTSA-N ampicillin Chemical compound C1([C@@H](N)C(=O)N[C@H]2[C@H]3SC([C@@H](N3C2=O)C(O)=O)(C)C)=CC=CC=C1 AVKUERGKIZMTKX-NJBDSQKTSA-N 0.000 description 1
- 230000003466 anti-cipated effect Effects 0.000 description 1
- 238000011861 anti-inflammatory therapy Methods 0.000 description 1
- 230000005875 antibody response Effects 0.000 description 1
- 210000000612 antigen-presenting cell Anatomy 0.000 description 1
- 230000000890 antigenic effect Effects 0.000 description 1
- 239000004599 antimicrobial Substances 0.000 description 1
- 230000000338 anxiogenic effect Effects 0.000 description 1
- PYMYPHUHKUWMLA-UHFFFAOYSA-N arabinose Natural products OCC(O)C(O)C(O)C=O PYMYPHUHKUWMLA-UHFFFAOYSA-N 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 230000002238 attenuated effect Effects 0.000 description 1
- 210000003719 b-lymphocyte Anatomy 0.000 description 1
- 210000000649 b-lymphocyte subset Anatomy 0.000 description 1
- SRBFZHDQGSBBOR-UHFFFAOYSA-N beta-D-Pyranose-Lyxose Natural products OC1COC(O)C(O)C1O SRBFZHDQGSBBOR-UHFFFAOYSA-N 0.000 description 1
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
- 229940004120 bifidobacterium infantis Drugs 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 210000004204 blood vessel Anatomy 0.000 description 1
- 230000037396 body weight Effects 0.000 description 1
- 239000006172 buffering agent Substances 0.000 description 1
- 239000006227 byproduct Substances 0.000 description 1
- 235000019577 caloric intake Nutrition 0.000 description 1
- 230000000711 cancerogenic effect Effects 0.000 description 1
- 231100000315 carcinogenic Toxicity 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 230000024245 cell differentiation Effects 0.000 description 1
- 239000006285 cell suspension Substances 0.000 description 1
- 230000010307 cell transformation Effects 0.000 description 1
- 239000002975 chemoattractant Substances 0.000 description 1
- 230000035605 chemotaxis Effects 0.000 description 1
- 229960005091 chloramphenicol Drugs 0.000 description 1
- WIIZWVCIJKGZOK-RKDXNWHRSA-N chloramphenicol Chemical compound ClC(Cl)C(=O)N[C@H](CO)[C@H](O)C1=CC=C([N+]([O-])=O)C=C1 WIIZWVCIJKGZOK-RKDXNWHRSA-N 0.000 description 1
- 230000019771 cognition Effects 0.000 description 1
- 230000001149 cognitive effect Effects 0.000 description 1
- 230000001332 colony forming effect Effects 0.000 description 1
- 238000004891 communication Methods 0.000 description 1
- OMFXVFTZEKFJBZ-HJTSIMOOSA-N corticosterone Chemical compound O=C1CC[C@]2(C)[C@H]3[C@@H](O)C[C@](C)([C@H](CC4)C(=O)CO)[C@@H]4[C@@H]3CCC2=C1 OMFXVFTZEKFJBZ-HJTSIMOOSA-N 0.000 description 1
- 239000006071 cream Substances 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 230000001186 cumulative effect Effects 0.000 description 1
- 238000011461 current therapy Methods 0.000 description 1
- 230000034994 death Effects 0.000 description 1
- 238000012217 deletion Methods 0.000 description 1
- 230000037430 deletion Effects 0.000 description 1
- 210000004443 dendritic cell Anatomy 0.000 description 1
- 230000007267 depressive like behavior Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 230000000378 dietary effect Effects 0.000 description 1
- 230000003292 diminished effect Effects 0.000 description 1
- 238000012377 drug delivery Methods 0.000 description 1
- 210000003162 effector t lymphocyte Anatomy 0.000 description 1
- 230000037149 energy metabolism Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 239000003623 enhancer Substances 0.000 description 1
- 210000002919 epithelial cell Anatomy 0.000 description 1
- ZMMJGEGLRURXTF-UHFFFAOYSA-N ethidium bromide Chemical compound [Br-].C12=CC(N)=CC=C2C2=CC=C(N)C=C2[N+](CC)=C1C1=CC=CC=C1 ZMMJGEGLRURXTF-UHFFFAOYSA-N 0.000 description 1
- 238000013401 experimental design Methods 0.000 description 1
- 210000002744 extracellular matrix Anatomy 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 229960002464 fluoxetine Drugs 0.000 description 1
- 235000012041 food component Nutrition 0.000 description 1
- 239000005428 food component Substances 0.000 description 1
- 235000012631 food intake Nutrition 0.000 description 1
- 229930182830 galactose Natural products 0.000 description 1
- 239000000499 gel Substances 0.000 description 1
- 238000012239 gene modification Methods 0.000 description 1
- 235000021474 generally recognized As safe (food) Nutrition 0.000 description 1
- 235000021473 generally recognized as safe (food ingredients) Nutrition 0.000 description 1
- 230000002068 genetic effect Effects 0.000 description 1
- 238000010353 genetic engineering Methods 0.000 description 1
- 230000005017 genetic modification Effects 0.000 description 1
- 235000013617 genetically modified food Nutrition 0.000 description 1
- 231100000024 genotoxic Toxicity 0.000 description 1
- 230000001738 genotoxic effect Effects 0.000 description 1
- 229960002518 gentamicin Drugs 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- AWUCVROLDVIAJX-UHFFFAOYSA-N glycerol 1-phosphate Chemical compound OCC(O)COP(O)(O)=O AWUCVROLDVIAJX-UHFFFAOYSA-N 0.000 description 1
- 239000008187 granular material Substances 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 230000007407 health benefit Effects 0.000 description 1
- 230000008821 health effect Effects 0.000 description 1
- 210000001320 hippocampus Anatomy 0.000 description 1
- 210000003630 histaminocyte Anatomy 0.000 description 1
- 230000013632 homeostatic process Effects 0.000 description 1
- 102000052624 human CXCL8 Human genes 0.000 description 1
- 102000052301 human GNAZ Human genes 0.000 description 1
- 102000043557 human IFNG Human genes 0.000 description 1
- 102000052620 human IL10 Human genes 0.000 description 1
- 102000057041 human TNF Human genes 0.000 description 1
- 229940116886 human interleukin-6 Drugs 0.000 description 1
- 229960000890 hydrocortisone Drugs 0.000 description 1
- 208000006575 hypertriglyceridemia Diseases 0.000 description 1
- 230000002631 hypothermal effect Effects 0.000 description 1
- 210000002865 immune cell Anatomy 0.000 description 1
- 230000036737 immune function Effects 0.000 description 1
- 230000003053 immunization Effects 0.000 description 1
- 230000007233 immunological mechanism Effects 0.000 description 1
- 230000007365 immunoregulation Effects 0.000 description 1
- 230000003308 immunostimulating effect Effects 0.000 description 1
- 230000001976 improved effect Effects 0.000 description 1
- 238000010348 incorporation Methods 0.000 description 1
- 239000000411 inducer Substances 0.000 description 1
- 230000002458 infectious effect Effects 0.000 description 1
- 230000008595 infiltration Effects 0.000 description 1
- 238000001764 infiltration Methods 0.000 description 1
- 210000004969 inflammatory cell Anatomy 0.000 description 1
- 239000003112 inhibitor Substances 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 230000015788 innate immune response Effects 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 229940096397 interleukin-8 Drugs 0.000 description 1
- 239000000543 intermediate Substances 0.000 description 1
- 210000001596 intra-abdominal fat Anatomy 0.000 description 1
- 210000005024 intraepithelial lymphocyte Anatomy 0.000 description 1
- 230000009545 invasion Effects 0.000 description 1
- 238000011813 knockout mouse model Methods 0.000 description 1
- 229940017800 lactobacillus casei Drugs 0.000 description 1
- NRYBAZVQPHGZNS-ZSOCWYAHSA-N leptin Chemical compound O=C([C@H](CO)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@H](CC=1C2=CC=CC=C2NC=1)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CO)NC(=O)CNC(=O)[C@H](CCC(N)=O)NC(=O)[C@@H](N)CC(C)C)CCSC)N1CCC[C@H]1C(=O)NCC(=O)N[C@@H](CS)C(O)=O NRYBAZVQPHGZNS-ZSOCWYAHSA-N 0.000 description 1
- 229940039781 leptin Drugs 0.000 description 1
- 231100000518 lethal Toxicity 0.000 description 1
- 230000001665 lethal effect Effects 0.000 description 1
- 229940054157 lexapro Drugs 0.000 description 1
- 230000037356 lipid metabolism Effects 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 235000015263 low fat diet Nutrition 0.000 description 1
- 238000002794 lymphocyte assay Methods 0.000 description 1
- 229960000274 lysozyme Drugs 0.000 description 1
- 235000010335 lysozyme Nutrition 0.000 description 1
- 239000004325 lysozyme Substances 0.000 description 1
- 230000003211 malignant effect Effects 0.000 description 1
- 230000001071 malnutrition Effects 0.000 description 1
- 235000000824 malnutrition Nutrition 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 230000001404 mediated effect Effects 0.000 description 1
- 230000002503 metabolic effect Effects 0.000 description 1
- 230000010034 metabolic health Effects 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 229960003085 meticillin Drugs 0.000 description 1
- 229960000282 metronidazole Drugs 0.000 description 1
- VAOCPAMSLUNLGC-UHFFFAOYSA-N metronidazole Chemical compound CC1=NC=C([N+]([O-])=O)N1CCO VAOCPAMSLUNLGC-UHFFFAOYSA-N 0.000 description 1
- 244000000010 microbial pathogen Species 0.000 description 1
- 230000002906 microbiologic effect Effects 0.000 description 1
- 239000003094 microcapsule Substances 0.000 description 1
- 244000005706 microflora Species 0.000 description 1
- 230000001483 mobilizing effect Effects 0.000 description 1
- 230000004899 motility Effects 0.000 description 1
- 210000004877 mucosa Anatomy 0.000 description 1
- 210000004400 mucous membrane Anatomy 0.000 description 1
- 231100000350 mutagenesis Toxicity 0.000 description 1
- 230000035772 mutation Effects 0.000 description 1
- OHDXDNUPVVYWOV-UHFFFAOYSA-N n-methyl-1-(2-naphthalen-1-ylsulfanylphenyl)methanamine Chemical compound CNCC1=CC=CC=C1SC1=CC=CC2=CC=CC=C12 OHDXDNUPVVYWOV-UHFFFAOYSA-N 0.000 description 1
- 229960000210 nalidixic acid Drugs 0.000 description 1
- MHWLWQUZZRMNGJ-UHFFFAOYSA-N nalidixic acid Chemical compound C1=C(C)N=C2N(CC)C=C(C(O)=O)C(=O)C2=C1 MHWLWQUZZRMNGJ-UHFFFAOYSA-N 0.000 description 1
- 230000031990 negative regulation of inflammatory response Effects 0.000 description 1
- 238000006386 neutralization reaction Methods 0.000 description 1
- 230000003472 neutralizing effect Effects 0.000 description 1
- 210000000440 neutrophil Anatomy 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 230000002474 noradrenergic effect Effects 0.000 description 1
- 229960002950 novobiocin Drugs 0.000 description 1
- YJQPYGGHQPGBLI-KGSXXDOSSA-N novobiocin Chemical compound O1C(C)(C)[C@H](OC)[C@@H](OC(N)=O)[C@@H](O)[C@@H]1OC1=CC=C(C(O)=C(NC(=O)C=2C=C(CC=C(C)C)C(O)=CC=2)C(=O)O2)C2=C1C YJQPYGGHQPGBLI-KGSXXDOSSA-N 0.000 description 1
- 208000015380 nutritional deficiency disease Diseases 0.000 description 1
- 229920001542 oligosaccharide Polymers 0.000 description 1
- 150000002482 oligosaccharides Chemical class 0.000 description 1
- 238000003305 oral gavage Methods 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 230000000242 pagocytic effect Effects 0.000 description 1
- 230000008506 pathogenesis Effects 0.000 description 1
- 239000008188 pellet Substances 0.000 description 1
- 229940056360 penicillin g Drugs 0.000 description 1
- 230000008855 peristalsis Effects 0.000 description 1
- 210000001986 peyer's patch Anatomy 0.000 description 1
- 239000008363 phosphate buffer Substances 0.000 description 1
- 239000006187 pill Substances 0.000 description 1
- 239000013612 plasmid Substances 0.000 description 1
- 238000007747 plating Methods 0.000 description 1
- 230000004983 pleiotropic effect Effects 0.000 description 1
- 229930001119 polyketide Natural products 0.000 description 1
- 150000003881 polyketide derivatives Chemical class 0.000 description 1
- 239000003910 polypeptide antibiotic agent Substances 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 238000010837 poor prognosis Methods 0.000 description 1
- 239000013641 positive control Substances 0.000 description 1
- 230000003389 potentiating effect Effects 0.000 description 1
- 231100000586 procarcinogen Toxicity 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 230000000069 prophylactic effect Effects 0.000 description 1
- 208000020016 psychiatric disease Diseases 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 239000003306 quinoline derived antiinfective agent Substances 0.000 description 1
- 239000012048 reactive intermediate Substances 0.000 description 1
- 230000025053 regulation of cell proliferation Effects 0.000 description 1
- 230000024833 regulation of cytokine production Effects 0.000 description 1
- 230000009892 regulation of energy homeostasis Effects 0.000 description 1
- 230000000241 respiratory effect Effects 0.000 description 1
- 229960003292 rifamycin Drugs 0.000 description 1
- HJYYPODYNSCCOU-ODRIEIDWSA-N rifamycin SV Chemical compound OC1=C(C(O)=C2C)C3=C(O)C=C1NC(=O)\C(C)=C/C=C/[C@H](C)[C@H](O)[C@@H](C)[C@@H](O)[C@@H](C)[C@H](OC(C)=O)[C@H](C)[C@@H](OC)\C=C\O[C@@]1(C)OC2=C3C1=O HJYYPODYNSCCOU-ODRIEIDWSA-N 0.000 description 1
- 201000000980 schizophrenia Diseases 0.000 description 1
- 230000003248 secreting effect Effects 0.000 description 1
- 230000028327 secretion Effects 0.000 description 1
- 230000035939 shock Effects 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 230000004936 stimulating effect Effects 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 239000002438 stress hormone Substances 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 239000000829 suppository Substances 0.000 description 1
- 230000002459 sustained effect Effects 0.000 description 1
- 235000019722 synbiotics Nutrition 0.000 description 1
- 208000011580 syndromic disease Diseases 0.000 description 1
- 239000006188 syrup Substances 0.000 description 1
- 235000020357 syrup Nutrition 0.000 description 1
- 201000000596 systemic lupus erythematosus Diseases 0.000 description 1
- 229960002180 tetracycline Drugs 0.000 description 1
- 229930101283 tetracycline Natural products 0.000 description 1
- 235000019364 tetracycline Nutrition 0.000 description 1
- 150000003522 tetracyclines Chemical class 0.000 description 1
- 230000017423 tissue regeneration Effects 0.000 description 1
- 239000003053 toxin Substances 0.000 description 1
- 231100000765 toxin Toxicity 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 238000011830 transgenic mouse model Methods 0.000 description 1
- -1 troches Substances 0.000 description 1
- 210000004881 tumor cell Anatomy 0.000 description 1
- 230000004614 tumor growth Effects 0.000 description 1
- 241001515965 unidentified phage Species 0.000 description 1
- 229960004688 venlafaxine Drugs 0.000 description 1
- PNVNVHUZROJLTJ-UHFFFAOYSA-N venlafaxine Chemical compound C1=CC(OC)=CC=C1C(CN(C)C)C1(O)CCCCC1 PNVNVHUZROJLTJ-UHFFFAOYSA-N 0.000 description 1
- 210000001835 viscera Anatomy 0.000 description 1
- 230000003442 weekly effect Effects 0.000 description 1
- 239000002676 xenobiotic agent Substances 0.000 description 1
- 230000002034 xenobiotic effect Effects 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
- C12N1/205—Bacterial isolates
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
- A23C9/00—Milk preparations; Milk powder or milk powder preparations
- A23C9/12—Fermented milk preparations; Treatment using microorganisms or enzymes
- A23C9/123—Fermented milk preparations; Treatment using microorganisms or enzymes using only microorganisms of the genus lactobacteriaceae; Yoghurt
- A23C9/1234—Fermented milk preparations; Treatment using microorganisms or enzymes using only microorganisms of the genus lactobacteriaceae; Yoghurt characterised by using a Lactobacillus sp. other than Lactobacillus Bulgaricus, including Bificlobacterium sp.
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23G—COCOA; COCOA PRODUCTS, e.g. CHOCOLATE; SUBSTITUTES FOR COCOA OR COCOA PRODUCTS; CONFECTIONERY; CHEWING GUM; ICE-CREAM; PREPARATION THEREOF
- A23G9/00—Frozen sweets, e.g. ice confectionery, ice-cream; Mixtures therefor
- A23G9/32—Frozen sweets, e.g. ice confectionery, ice-cream; Mixtures therefor characterised by the composition containing organic or inorganic compounds
- A23G9/36—Frozen sweets, e.g. ice confectionery, ice-cream; Mixtures therefor characterised by the composition containing organic or inorganic compounds containing microorganisms or enzymes; containing paramedical or dietetical agents, e.g. vitamins
- A23G9/363—Frozen sweets, e.g. ice confectionery, ice-cream; Mixtures therefor characterised by the composition containing organic or inorganic compounds containing microorganisms or enzymes; containing paramedical or dietetical agents, e.g. vitamins containing microorganisms, enzymes
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/135—Bacteria or derivatives thereof, e.g. probiotics
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/66—Microorganisms or materials therefrom
- A61K35/74—Bacteria
- A61K35/741—Probiotics
- A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
- A61K35/745—Bifidobacteria
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/02—Stomatological preparations, e.g. drugs for caries, aphtae, periodontitis
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/04—Drugs for disorders of the alimentary tract or the digestive system for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/12—Antidiarrhoeals
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/16—Drugs for disorders of the alimentary tract or the digestive system for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P11/00—Drugs for disorders of the respiratory system
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P11/00—Drugs for disorders of the respiratory system
- A61P11/06—Antiasthmatics
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P13/00—Drugs for disorders of the urinary system
- A61P13/12—Drugs for disorders of the urinary system of the kidneys
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P15/00—Drugs for genital or sexual disorders; Contraceptives
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
- A61P17/02—Drugs for dermatological disorders for treating wounds, ulcers, burns, scars, keloids, or the like
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
- A61P17/06—Antipsoriatics
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
- A61P17/10—Anti-acne agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P19/00—Drugs for skeletal disorders
- A61P19/02—Drugs for skeletal disorders for joint disorders, e.g. arthritis, arthrosis
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P19/00—Drugs for skeletal disorders
- A61P19/08—Drugs for skeletal disorders for bone diseases, e.g. rachitism, Paget's disease
- A61P19/10—Drugs for skeletal disorders for bone diseases, e.g. rachitism, Paget's disease for osteoporosis
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P25/00—Drugs for disorders of the nervous system
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P25/00—Drugs for disorders of the nervous system
- A61P25/28—Drugs for disorders of the nervous system for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
- A61P3/04—Anorexiants; Antiobesity agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
- A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
- A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/04—Antibacterial agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/10—Antimycotics
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/12—Antivirals
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/12—Antivirals
- A61P31/14—Antivirals for RNA viruses
- A61P31/18—Antivirals for RNA viruses for HIV
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
- A61P35/04—Antineoplastic agents specific for metastasis
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
- A61P37/02—Immunomodulators
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
- A61P37/02—Immunomodulators
- A61P37/06—Immunosuppressants, e.g. drugs for graft rejection
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
- A61P37/08—Antiallergic agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P5/00—Drugs for disorders of the endocrine system
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P7/00—Drugs for disorders of the blood or the extracellular fluid
- A61P7/02—Antithrombotic agents; Anticoagulants; Platelet aggregation inhibitors
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P7/00—Drugs for disorders of the blood or the extracellular fluid
- A61P7/06—Antianaemics
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P9/00—Drugs for disorders of the cardiovascular system
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P9/00—Drugs for disorders of the cardiovascular system
- A61P9/10—Drugs for disorders of the cardiovascular system for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/51—Bifidobacterium
- A23V2400/533—Longum
-
- A23Y2300/55—
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Definitions
- the invention relates to a Bifidobacterium strain and its use as a probiotic bacteria in particular as an immunomodulatory biotherapeutic agent.
- the defense mechanisms to protect the human gastrointestinal tract from colonization by intestinal bacteria are highly complex and involve both immunological and non-immunological aspects (1).
- Innate defense mechanisms include the low pH of the stomach, bile salts, peristalsis, mucin layers and anti-microbial compounds such as lysozyme (2).
- Immunological mechanisms include specialized lymphoid aggregates, underlying M cells, called peyers patches which are distributed throughout the small intestine and colon (3). Luminal antigens presented at these sites result in stimulation of appropriate T and B cell subsets with establishment of cytokine networks and secretion of antibodies into the gastrointestinal tract (4).
- antigen presentation may occur via epithelial cells to intraepithelial lymphocytes and to the underlying lamina limba immune cells (5). Therefore, the host invests substantially in immunological defense of the gastrointestinal tract.
- the gastrointestinal mucosa is the largest surface at which the host interacts with the external environment, specific control mechanisms must be in place to regulate immune responsiveness to the 100 tons of food which is handled by the gastrointestinal tract over an average lifetime.
- the gut is colonized by over 500 species of bacteria numbering 10 11 -10 12 /g in the colon.
- these control mechanisms must be capable of distinguishing non-pathogenic adherent bacteria from invasive pathogens, which would cause significant damage to the host.
- the intestinal flora contributes to defense of the host by competing with newly ingested potentially pathogenic micro-organisms.
- Bacteria present in the human gastrointestinal tract can promote inflammation. Aberrant immune responses to the indigenous microflora have been implicated in certain disease states, such as inflammatory bowel disease. Antigens associated with the normal flora usually lead to immunological tolerance and failure to achieve this tolerance is a major mechanism of mucosal inflammation (6). Evidence for this breakdown in tolerance includes an increase in antibody levels directed against the gut flora in patients with inflammatory bowel disease (IBD).
- IBD inflammatory bowel disease
- the present invention is directed towards a Bifidobacterium strain which has been shown to have immunomodulatory effects, by modulating cytokine levels or by antagonizing and excluding pro-inflammatory micro-organisms from the gastrointestinal tract.
- the invention provides an isolated strain of Bifidobacterium NCIMB 41676.
- the Bifidobacterium strain may be in the form of viable cells.
- the Bifidobacterium strain may be in the form of non-viable cells.
- the Bifidobacterium may be isolated from colonic biopsy tissue from a healthy human subject.
- the Bifidobacterium strain may be significantly immunomodulatory following oral consumption in humans.
- the invention also provides a formulation which comprises a Bifidobacterium strain as described herein.
- the formulation may further comprise a probiotic material.
- the formulation may further comprise a prebiotic material.
- the formulation may further comprise an ingestable carrier.
- the ingestable carrier may a pharmaceutically acceptable carrier such as a capsule, tablet or powder.
- the ingestable carrier may be a food product such as acidified milk, yoghurt, frozen yoghurt, milk powder, milk concentrate, cheese spreads, dressings or beverages.
- the formulation may further comprise a protein and/or peptide, in particular proteins and/or peptides that are rich in glutamine/glutamate, a lipid, a carbohydrate, a vitamin, mineral and/or trace element.
- the Bifidobacterium strain may be present in an amount of more than 10 6 cfu per gram of the formulation.
- the formulation may further comprise an adjuvant.
- the formulation may further comprise a bacterial component.
- the formulation may further comprise a drug entity.
- the formulation may further comprise a biological compound.
- the formulation may be used for immunisation and vaccination protocols.
- the invention also provides a Bifidobacterium strain or a formulation as described herein for use in foodstuffs.
- the invention also provides a Bifidobacterium strain or a formulation as described herein for use as a medicament.
- the invention also provides a Bifidobacterium strain or a formulation as described herein for use in the prophylaxis and/or treatment of undesirable inflammatory activity.
- the invention also provides a Bifidobacterium strain or a formulation as described herein for use in the prophylaxis and/or treatment of undesirable gastrointestinal inflammatory activity such as inflammatory bowel disease eg. Crohns disease or ulcerative colitis, irritable bowel syndrome; pouchitis; or post infection colitis.
- undesirable gastrointestinal inflammatory activity such as inflammatory bowel disease eg. Crohns disease or ulcerative colitis, irritable bowel syndrome; pouchitis; or post infection colitis.
- the invention also provides a Bifidobacterium strain or a formulation as described herein for use in the prophylaxis and/or treatment of gastrointestinal cancer(s).
- the invention also provides a Bifidobacterium strain or a formulation as described herein for use in the prophylaxis and/or treatment of systemic disease such as rheumatoid arthritis.
- the invention also provides a Bifidobacterium strain or a formulation as described herein for use in the prophylaxis and/or treatment of autoimmune disorders due to undesirable inflammatory activity.
- the invention also provides a Bifidobacterium strain or a formulation as described herein for use in the prophylaxis and/or treatment of cancer due to undesirable inflammatory activity.
- the invention also provides a Bifidobacterium strain or a formulation as described herein for use in the prophylaxis of cancer.
- the invention also provides a Bifidobacterium strain or a formulation as described herein for use in the prophylaxis and/or treatment of diarrhoeal disease due to undesirable inflammatory activity, such as Clostridium difficile associated diarrhoea, Rotavirus associated diarrhoea or post infective diarrhoea or diarrhoeal disease due to an infectious agent, such as E. coli.
- undesirable inflammatory activity such as Clostridium difficile associated diarrhoea, Rotavirus associated diarrhoea or post infective diarrhoea or diarrhoeal disease due to an infectious agent, such as E. coli.
- the invention also provides a Bifidobacterium strain or a formulation as described herein for use in the preparation of anti-inflammatory biotherapeutic agents for the prophylaxis and/or treatment of undesirable inflammatory activity.
- Bifidobacterium strains as described herein may be used in the preparation of a panel of biotherapeutic agents for modifying the levels of IL-10.
- the invention also provides a Bifidobacterium strain or a formulation as described herein for use in the prevention and/or treatment of inflammatory disorders, immunodeficiency, inflammatory bowel disease, irritable bowel syndrome, cancer (particularly of the gastrointestinal and immune systems), diarrhoeal disease, antibiotic associated diarrhoea, paediatric diarrhoea, appendicitis, autoimmune disorders, multiple sclerosis, Alzheimer's disease, rheumatoid arthritis, coeliac disease, diabetes mellitus, organ transplantation, bacterial infections, viral infections, fungal infections, periodontal disease, urogenital disease, sexually transmitted disease, HIV infection, HIV replication, HIV associated diarrhoea, surgical associated trauma, surgical-induced metastatic disease, sepsis, weight loss, anorexia, fever control, cachexia, wound healing, ulcers, gut barrier function, allergy, asthma, respiratory disorders, circulatory disorders, coronary heart disease, anaemia, disorders of the blood coagulation system, renal
- the Bifidobacterium strain as described herein may act by antagonising and excluding proinflammatory micro-organisms from the gastrointestinal tract.
- the invention also provides a Bifidobacterium strain or a formulation as described herein for use in the preparation of anti-inflammatory biotherapeutic agents for reducing the levels of pro inflammatory cytokines.
- the Bifidobacterium strain as described herein may be used as an anti-infective probiotic strain.
- the invention also provides a Bifidobacterium strain or a formulation as described herein for use in the prophylaxis and/or treatment of bipolar illness, depression, mood disorders, and/or anxiety disorders.
- the invention also provides a Bifidobacterium strain or a formulation as described herein may be used as a cognative enhancer for the prophylaxis and/or treatment of disorders of the central nervous system such as Alzheimer's disease, schizophrenia and/or mild cognative disorder.
- the invention also provides a Bifidobacterium strain or a formulation as described herein for use in the prophylaxis and/or treatment of obesity related inflammation.
- the invention also provides a Bifidobacterium strain or a formulation as described herein for use in the prophylaxis and/or treatment of obesity related metabolic dysregulation.
- Bifidobacterium strain AH1714 (NCIMB 41676) or mutants or variants thereof.
- the mutant may be a genetically modified mutant.
- the variant may be a naturally occurring variant of Bifidobacterium .
- a rifampicin resistant variant of strain AH1714 is also described.
- the strain may be a probiotic. It may be in the form of a biologically pure culture.
- Bifidobacterium NCNB 41676 We also describe an isolated strain of Bifidobacterium NCNB 41676.
- the Bifidobacterium 30 strains may be in the form of viable cells.
- Bifidobacterium strains are in the form of non-viable cells.
- the general use of probiotic bacteria is in the form of viable cells.
- non-viable cells such as killed cultures or compositions containing beneficial factors expressed by the probiotic bacteria. This could include thermally killed micro-organisms or micro-organisms killed by exposure to altered pH or subjection to pressure or gamma irradiation.
- With non-viable cells product preparation is simpler, cells may be incorporated easily into pharmaceuticals and storage requirements are much less limited than viable cells.
- Lactobacillus casei YIT 9018 offers an example of the effective use of heat killed cells as a method for the treatment and/or prevention of tumour growth as described in U.S. Pat. No. 4,347,240.
- the Bifidobacterium strains may be isolated from colonic biopsy tissue from healthy human subjects, the Bifidobacterium strains being significantly immunomodulatory following oral consumption in humans.
- the formulation may include another probiotic material.
- the formulation may include a prebiotic material.
- the formulation includes an ingestable carrier.
- the ingestable carrier may be a pharmaceutically acceptable carrier such as a capsule, tablet or powder.
- the ingestable carrier is a food product such as acidified milk, yoghurt, frozen yoghurt, milk powder, milk concentrate, cheese spreads, dressings or beverages.
- the formulation may further comprise a protein and/or peptide, in particular proteins and/or peptides that are rich in glutamine/glutamate, a lipid, a carbohydrate, a vitamin, mineral and/or trace element.
- the Bifidobacterium strain may be present in the formulation at more than 10 6 cfu per gram of delivery system.
- the formulation includes any one or more of an adjuvant, a bacterial component, a drug entity or a biological compound.
- Bifidobacterium strain or a formulation for use as foodstuffs, as a medicament for use in the prophylaxis and/or treatment of undesirable inflammatory activity, for use in the prophylaxis and/or treatment of undesirable respiratory inflammatory activity such as asthma, for use in the prophylaxis and/or treatment of undesirable gastrointestinal inflammatory activity such as inflammatory bowel disease eg.
- Crohns disease or ulcerative colitis, irritable bowel syndrome, pouchitis, or post infection colitis for use in the prophylaxis and/or treatment of gastrointestinal cancer(s), for use in the prophylaxis and/or treatment of systemic disease such as rheumatoid arthritis, for use in the prophylaxis and/or treatment of autoimmune disorders due to undesirable inflammatory activity, for use in the prophylaxis and/or treatment of cancer due to undesirable inflammatory activity, for use in the prophylaxis of cancer, for use in the prophylaxis and/or treatment of diarrhoeal disease due to undesirable inflammatory activity, such as Clostridium difficile associated diarrhoea, Rotavirus associated diarrhoea or post infective diarrhoea, for use in the prophylaxis and/or treatment of diarrhoeal disease due to an infectious agent, such as E. coli.
- systemic disease such as rheumatoid arthritis
- Bifidobacterium strain or a formulation of the invention for use in the preparation of an anti-inflammatory biotherapeutic agent for the prophylaxis and/or treatment of undesirable inflammatory activity or for use in the preparation of anti-inflammatory biotherapeutic agents for the prophylaxis and/or treatment of undesirable inflammatory activity.
- the strain may act by antagonising and excluding proinflammatory micro-organisms from the gastrointestinal tract.
- the Bifidobacterium strain may be used in the preparation of anti-inflammatory biotherapeutic agents for modifying the levels of IL-10.
- the Bifidobacterium strain may be used as a anti-infective probiotic due to their ability to antagonise the growth of pathogenic species.
- the invention is therefore of major potential therapeutic value in the prophylaxis or treatment of dysregulated immune responses, such as undesirable inflammatory reactions for example asthma.
- Bifidobacterium are commensal microorganisms. They have been isolated from the microbial flora within the human gastrointestinal tract. The immune system within the gastrointestinal tract cannot have a pronounced reaction to members of this flora, as the resulting inflammatory activity would also destroy host cells and tissue function. Therefore, some mechanism(s) exist whereby the immune system can recognize commensal non-pathogenic members of the gastrointestinal flora as being different to pathogenic organisms. This ensures that damage to host tissues is restricted and a defensive barrier is still maintained.
- a deposit of Bifidobacterium longum strain AH1714 was made at the National Collections of Industrial and Marine Bacteria Limited (NCIMB) Ferguson Building, Craibstone Estate, Bucksburn, Aberdeen, AB21 9YA, Scotland, UK on Nov. 5, 2009 and accorded the accession number NCIMB 41676.
- NCIMB National Collections of Industrial and Marine Bacteria Limited
- the Bifidobacterium longum may be a genetically modified mutant or it may be a naturally occurring variant thereof.
- the Bifidobacterium longum may be in the form of viable cells.
- the Bifidobacterium longum may be in the form of non-viable cells.
- the specific Bifidobacterium strain of the invention may be administered to animals (including humans) in an orally ingestible form in a conventional preparation such as capsules, microcapsules, tablets, granules, powder, troches, pills, suppositories, suspensions and syrups.
- a conventional preparation such as capsules, microcapsules, tablets, granules, powder, troches, pills, suppositories, suspensions and syrups.
- Suitable formulations may be prepared by methods commonly employed using conventional organic and inorganic additives.
- the amount of active ingredient in the medical composition may be at a level that will exercise the desired therapeutic effect.
- the formulation may also include a bacterial component, a drug entity or a biological compound.
- a vaccine comprising the strains of the invention may be prepared using any suitable known method and may include a pharmaceutically acceptable carrier or adjuvant.
- mutant, variant and genetically modified mutant include a strain of Bifidobacteria whose genetic and/or phenotypic properties are altered compared to the parent strain.
- Naturally occurring variant of Bifidobacterium longum includes the spontaneous alterations of targeted properties selectively isolated.
- Deliberate alteration of parent strain properties is accomplished by conventional (in vitro) genetic manipulation technologies, such as gene disruption, conjugative transfer, etc.
- Genetic modification includes introduction of exogenous and/or endogenous DNA sequences into the genome of a Bifidobacteria strain, for example by insertion into the genome of the bacterial strain by vectors, including plasmid DNA, or bacteriophages.
- Natural or induced mutations include at least single base alterations such as deletion, insertion, transversion or other DNA modifications which may result in alteration of the amino acid sequence encoded by the DNA sequence.
- mutant, variant and genetically modified mutant also include a strain of Bifidobacteria that has undergone genetic alterations that accumulate in a genome at a rate which is consistent in nature for all micro-organisms and/or genetic alterations which occur through spontaneous mutation and/or acquisition of genes and/or loss of genes which is not achieved by deliberate (in vitro) manipulation of the genome but is achieved through the natural selection of variants and/or mutants that provide a selective advantage to support the survival of the bacterium when exposed to environmental pressures such as antibiotics.
- a mutant can be created by the deliberate (in vitro) insertion of specific genes into the genome which do not fundamentally alter the biochemical functionality of the organism but whose products can be used for identification or selection of the bacterium, for example antibiotic resistance.
- mutant or variant strains of Bifidobacteria can be identified by DNA sequence homology analysis with the parent strain. Strains of Bifidobacteria having a close sequence identity with the parent strain are considered to be mutant or variant strains. A Bifidobacteria strain with a sequence identity (homology) of 96% or more, such as 97% or more, or 98% or more, or 99% or more with the parent DNA sequence may be considered to be a mutant or variant. Sequence homology may be determined using on-line homology algorithm “BLAST” program, publicly available at http://www.ncbi.nlm.nih.gov/BLAST/.
- Mutants of the parent strain also include derived Bifidobacteria strains having at least 85% sequence homology, such as at least 90% sequence homology, or at least 95% sequence homology to the 16s-23s intergenic spacer polynucleotide sequence of the parent strain. These mutants may further comprise DNA mutations in other DNA sequences in the bacterial genome.
- FIG. 1 is a graph illustrating transit of B. longum AH1714 through the gastrointestinal tract
- FIG. 2 is a photograph of B. longum AH1714 grown on a Congo Red Agar plate
- FIG. 3 is a bar chart illustrating the IL-10:IL-12p70 ratio for PBMCs stimulated with Bifidobacterium longum strain 1714 ( Bifidobacterium 1714);
- FIG. 4 is a bar chart showing the induction profile of IL-10 in splenocytes isolated from both 1714 and PBS fed mice with and without in vivo LPS challenge 1 mg/kg.
- In vitro cells are either unstimulated (A), stimulated with LPS (B) or stimulated with antiCD3/CD28 (C). Data is shown as Average & SEM;
- FIG. 5 is a bar chart showing the induction profile of TNF- ⁇ in splenocytes isolated from both 1714 and PBS fed mice with and without in vivo LPS challenge 1 mg/kg.
- In vitro cells are either unstimulated (A) or stimulated with antiCD3/CD28 (B). Data is shown as Average & SEM;
- FIG. 6 is a bar chart showing the induction profile of IFN- ⁇ in splenocytes isolated from both 1714 and PBS fed mice with and without in vivo LPS challenge 1 mg/kg.
- In vitro cells are either unstimulated (A) or stimulated with antiCD3/CD28 (B). Data is shown as Average & SEM.
- FIG. 7 is a bar chart showing the induction profile of IL-12p70 in splenocytes isolated from both 1714 and PBS fed mice with and without in vivo LPS challenge 1 mg/kg.
- In vitro cells are either unstimulated (A) or stimulated with antiCD3/CD28 (B). Data is shown as Average & SEM;
- FIG. 8 is a bar chart showing the induction profile of TNF- ⁇ (A) and IL-10 (B) in serum sampled from both 1714 and PBS fed mice post 2H in vivo challenge with LPS 1 mg/kg. Data is shown as Average & SEM;
- FIG. 9 is a bar chart showing NFkB activity (Photons/second) from isolated spleen 3 hours post challenge with a single 0.5 mg/kg dose of LPS, from Placebo and 1714-fed animals (** designates p ⁇ 0.01);
- FIG. 10 is a bar chart (A) showing NFkB activity (Photons/second) from whole body imaging 1.5 hours post challenge with a single 0.5 mg/kg dose of LPS, from Placebo and 1714-fed animals ((B) and (C) are whole body representative images in black and white and colour;
- FIG. 11 is a bar chart representing the time of immobility displayed by the mice over a 6-min test
- FIG. 12 is a line graph representing the freezing percentage in response to the fearful stimulus (context) for day 1 (acquisition), day 2 (memory/extinction) and day 3 (extinction);
- FIG. 13 is a line graph representing the freezing percentage in response to the fearful stimulus (cue) for day 1 (acquisition), day 2 (memory/extinction) and day 3 (extinction);
- FIG. 14 is a bar chart representing the number of marbles buried by the mice over a 30-min session
- FIG. 15 is a bar chart representing the body temperature variation ( ⁇ T) mice displayed following handling.
- FIG. 16 is a bar chart showing the changes in cytokine levels in stimulated splenocytes from the Diet Induced Mouse model.
- a deposit of Bifidobacterium longum strain AH1714 was made at the National Collections of Industrial and Marine Bacteria Limited (NCIMB) Ferguson Building, Craibstone Estate, Bucksburn, Aberdeen, AB21 9YA, Scotland, UK on Nov. 5, 2009 and accorded the accession number NCIMB 41676.
- NCIMB National Collections of Industrial and Marine Bacteria Limited
- a deposit of Bifidobacterium longum strain UCC35624 was made at the National Collections of Industrial and Marine Bacteria Limited (NCIMB) Ferguson Building, Craibstone Estate, Bucksburn, Aberdeen, AB21 9YA, Scotland, UK on Jan. 13, 1999 and accorded the accession number NCIMB 41003.
- NCIMB National Collections of Industrial and Marine Bacteria Limited
- Bifidobacterium longum strain AH1714 was isolated from colonic biopsy tissue from healthy human subjects.
- PBS Phosphate Buffered Saline
- Triton X-100 (0.05%) was added to release the adherent microorganisms from the tissue sample. Tissue samples were then incubated for 10 min.
- the samples were vortexed vigorously and adherent Lactobacilli and Bifidobacteria isolated from the gastrointestinal tissue by plating on selective agar (De Man, Rogosa and Sharpe (MRS) agar+Vancomycin and Wilkins-Chalgren Agar+Mupirocin, respectively). Isolated colonies were picked from the plates and re-streaked three times to ensure purity. Microscope examination, Gram staining, Catalase testing, Fructose-6-Phosphate Phosphoketolase assessment were used to determine presumptive Bifidobacteria species and isolates were stocked in 40% glycerol and stored at ⁇ 20° and ⁇ 80° C. 16S intergenic spacer region sequencing were used to confirm the identity of the newly isolated strains.
- AH1714 is a gram positive, catalase negative pleomorphic shaped bacterium which is Fructose-6-Phosphate Phosphoketolase positive, confirming its identity as a bifidobacterium .
- IGS intergenic spacer
- the cycling conditions were 94° C. for 4 min (1 cycle), 94° C. for 45 sec, 53° C. for 45 sec, 72° C. for 45 sec (28 cycles).
- the PCR reaction contained 2 ⁇ l (100 ng) of DNA, PCR mix (Sigma, Red Taq), 0.025 nM IGS L and R primer (MWG Biotech, Germany). The PCR reactions were performed on a Biotherma thermocycler. The PCR products (10 ⁇ l) were ran alongside a molecular weight marker (100 bp Ladder, Roche) on a 2% agarose EtBr stained gel in TAE, to determine the IGS profile.
- a molecular weight marker 100 bp Ladder, Roche
- PCR products of Bifidobacterium were purified using the Promega Wizard PCR purification kit.
- the purified PCR products were sequenced using the primer sequences (above) for the intergenic spacer region. Sequence data was then searched against the NCBI nucleotide database to determine the identity of the strain by nucleotide homology.
- the resultant DNA sequence data was subjected to the NCBI standard nucleotide-to-nucleotide homology BLAST search engine (http://www.ncbi.nlm.nih.gov/BLAST/). The nearest match to the sequence was identified and then the sequences were aligned for comparison using DNASTAR MegAlign software.
- the sequences (SEQ ID NO.
- PCR was performed using BOX primers (8).
- the cycling conditions were 94° C. for 7 min (1 cycle); 94° C. for 1 minute, 53° C. for 45 secs, 65° C. for 8 minutes, (30 cycles) and 65° C. for 16 minutes.
- the PCR reaction contained 50 ng of DNA, PCR mix (Sigma, Red Taq) and 0.03 nM BOXA1R primer (5′-CTACGGCAAGGCGACGCTGACG-3′) (SEQ ID NO. 5) (MWG Biotech, Germany).
- the PCR reactions were performed on a Biotherma thermocycler.
- the PCR products were run on a 3% agarose gel alongside a molecular weight marker (Roche, 100 bp ladder) and imaged.
- the antibiotic sensitivity profiles for B. longum AH1714 was determined using the ‘disc susceptibility’ assay.
- the cultures were grown up in the appropriate broth medium for 48 h and spread-plated (100 ⁇ l) onto agar media. Discs containing known concentrations of the antibiotics were placed onto the agar. Strains were examined for antibiotic sensitivity after 1-2 days incubation at 37° C. under anaerobic conditions.
- AH1714 To determine whether Bifidobacterium longum AH1714 could survive at low pH values equivalent to those found in the stomach, bacterial cells were harvested from fresh overnight cultures, washed twice in phosphate buffer (pH 6.5) and resuspended in TPY broth adjusted to pH 2.5 (with 1M HCl). Cells were incubated at 37° C. and survival measured at intervals of 5, 30, 60 and 120 minutes using the plate count method. AH1714 survived well for 5 minutes at pH 2.5 while no viable cells were recovered after 30 minutes.
- AH1714 was grown overnight in MRS (supplemented with 0.05% cysteine-HCl). 2 ⁇ l of AH1714 culture was spotted onto the agar and incubated for 24 h.
- the indicator organisms were grown in TSB ( E. coli and Salmonella typhimurium ), Brucella broth ( Campylobacter jejuni ) and Reinforced Clostridia Media (RCM, Clostridium difficile ).
- the indicator lawn was prepared by inoculating a molten overlay with 2% 0.5 (v/v) of the overnight indicator culture, which was poured onto the surface of the spotted probiotic cultures following overnight growth on the agar plates. Plates were incubated at 37° C. under suitable conditions for the indicator strain and the growth recorded after 24-48 hr. Zones of clearing greater than 1 mm diameter were considered sensitive to the probiotic strain.
- This assay was also performed on media supplemented with 2% 3-glycerophosphate as a buffering agent to limit antagonistic activity due to acid production.
- a spontaneous rifampicin-resistant variant (rif+) was isolated as follows: a fresh broth culture of AH1714 was spread-plated (100 ⁇ L) onto MRS+rifampicin+cysteine with the lowest concentration of rifampicin (range was 0.1%, 0.08%, 0.06%, 0.04%, 0.02% and 0.002%). Plate media without rifampicin was included as a positive control. Both sets of plates were incubated anaerobically at 37° C. (48 hours).
- the removed plates were assessed for purity before picking one colony from the rifampicin supplemented agar plate and streaking onto the rifampicin supplemented plate of next highest concentration.
- a colony was streaked from the MRS agar plate onto a fresh MRS agar plate and both sets of plates incubated anaerobically at 37° C. (48 hours). This process was repeated for the full range of rifampicin supplemented plates.
- a single colony from a fully grown culture on a 50 ⁇ g/mL rifampicin supplemented MRS agar plate was used to inoculate into 20 ml MRS broth and the resultant culture used for subsequent stocking. The identity of the variant was confirmed by microscopic assessment, IGS sequence analysis and by specific PCR analysis.
- a Congo red agar screen was used to phenotypically screen for EPS expressing bacterial strains. Briefly, 10 ml Modified Rogosa broth media (+0.05% cysteine) was inoculated aseptically with a freshly grown colony of the bacterial strain and incubated anaerobically at 37° C. until turbid (about 16 to about 24 hours). The broth cultures were aseptically streaked onto Congo Red Agar plates and incubated anaerobically at 37° C. for 48 hours. It is believed that EPS produced as a by-product of the growth and/or metabolism of certain strains prevents the uptake of the Congo red stain resulting in a cream/white colony morphology. Stains that produce less EPS take up the Congo red stain easily, resulting in a pink/red colony morphology. Strains that do not produce an EPS stain red and look almost transparent in the red agar background.
- the colony morphology for B. longum AH1714 is convex, mucoid, bright white colonies
- Example 3 Bacillus subtilis 1714 Induces a Significantly Elevated IL-10:IL-12 Ratio
- PBMCs Peripheral blood mononuclear cells
- BD Vacutainer CPT tubes BD catalog 362761
- PBMCs were washed and resuspended in Dulbecco's Modified Eagle Medium-GlutamaxTM (Glutamax (Glutamine substitute)+pyruvate+4.5 g/l glucose (Gibco catalog 10569-010) 10% fetal bovine serum (Sigma catalog F4135), and 1% penicillin/streptomycin (Sigma catalog P0781).
- Dulbecco's Modified Eagle Medium-GlutamaxTM Glutamax (Glutamine substitute)+pyruvate+4.5 g/l glucose (Gibco catalog 10569-010) 10% fetal bovine serum (Sigma catalog F4135), and 1% penicillin/streptomycin (Sigma catalog P0781).
- PBMCs were incubated (2 ⁇ 10 5 cells per well) in flat-bottomed 96-well plates and 20 ⁇ L of a bacterial suspension (at a concentration of 1 ⁇ 10 7 CFU/mL) was added. PBMCs were co-incubated with bacteria for 48 hours at 37° C./5% CO 2 in an incubator. After the 2 day incubation period, the plates were centrifuged at 300 ⁇ g, and the supernatants were removed and stored frozen at ⁇ 80° C. until analysis.
- Interleukin-10 IL-10
- Interleukin-12p70 IL-12p70
- Bacteria were prepared for co-culture experiments in two formats. (a) Freshly grown bacteria were grown in Difco MRS media and harvested just after entering into stationary phase. All cells were grown under anaerobic conditions at 37° C. (b) Bacteria were grown under anaerobic conditions at 37° C. in Difco MRS media and harvested just after entering into stationary phase. Freeze dried powders were generated for each of these bacteria and stored at ⁇ 80° C. in pre-aliquoted 100 mg vials. Immediately prior to their use, one aliquot of each strain was removed from the freezer and allowed to reach room temperature. Each strain was washed 3 times in 10 ml ringers followed by centrifugation. A fresh vial was used on each occasion.
- the control of inflammatory diseases is exerted at a number of levels.
- the controlling factors include hormones, prostaglandins, reactive oxygen and nitrogen intermediates, leukotrienes and cytokines.
- Cytokines are low molecular weight biologically active proteins that are involved in the generation and control of immunological and inflammatory responses. A number of cell types produce these cytokines, with neutrophils, monocytes and lymphocytes being the major sources during inflammatory reactions due to their large numbers at the injured site.
- cytokines generated at inflammatory sites influence the inflammatory response.
- Chemotaxis stimulates homing of inflammatory cells to the injured site, whilst certain cytokines promote infiltration of cells into tissue. Cytokines released within the injured tissue result in activation of the inflammatory infiltrate. Most cytokines are pleiotropic and express multiple biologically overlapping activities. As uncontrolled inflammatory responses can result in diseases such as IBD, it is reasonable to expect that cytokine production has gone astray in individuals affected with these diseases.
- Interleukin-10 is an anti-inflammatory cytokine which is produced by many cell types including monocytes, macrophages, dendritic cells, mast cells and lymphocytes (in particular T regulatory cells). IL-10 down-regulates the expression of pro-inflammatory Th1 cytokines, MHC class II antigens, and co-stimulatory molecules on antigen presenting cells. It also enhances B cell survival, proliferation, and antibody production. This cytokine can block NF- ⁇ B activity, and is involved in the regulation of the JAK-STAT signaling pathway. Murine knock-out studies have demonstrated the essential role for IL-10 in immunoregulation as IL-10KO mice develop severe colitis.
- Interleukin-12 is a pro-inflammatory cytokine associated with polarisation of Th1 effector T cell responses and stimulates the production of other pro-inflammatory Th1 cytokines, such as interferon-gamma (IFN- ⁇ ) and tumor necrosis factor-alpha (TNF- ⁇ ), from T and natural killer (NK) cells.
- IFN- ⁇ interferon-gamma
- TNF- ⁇ tumor necrosis factor-alpha
- NK natural killer cells.
- High levels of IL-12 expression is associated with autoimmunity.
- Administration of IL-12 to people suffering from autoimmune diseases was shown to worsen disease symptoms.
- IL-12 knock-out mice or treatment of mice with IL-12 neutralising antibodies ameliorated the disease.
- Cytokine cascades and networks control the inflammatory response, rather than the action of a particular cytokine on a particular cell type.
- the relative levels of expression, or balance, of two cytokines is more informative than the expression of a single cytokine.
- PBMC IL-10:IL-12 ratio is a important selection criterion for identification of bacterial strains with immunoregulatory properties.
- Example 4 Long Term Feeding of Mice with Bif. AH1714 is Associated with Increased Anti-Inflammatory Cytokine IL-10 and with Decreased Pro-Inflammatory and Th1 Cytokines TNF-IFN- ⁇ and IL-12 in Healthy Animals and in a Model of Sepsis/Inflammation
- mice @ 6-8 weeks of age are sourced from Harlan UK and housed in individually ventilated cages and provided ad libitum access to sterile standard mouse chow and water.
- a further 10 AH1714 mice and 6 control mice are administered a single dose of LPS @ 1 mg/kg via intraperitoneal injection. After 2 hours blood is sampled and the mice were culled. Serum and splenocyte cells were treated and analysed as previously described.
- Splenocytes are isolated from spleens and incubated for 48 hours at 37° C. (in the presence of penicillin and streptomycin) with control media, LPS, or antiCD3/CD28. Cytokines in the culture supernatants are assayed using a 96-well assay kit from Meso Scale Discovery (Gaithersburg, Md.; catalog K15008B-1).
- Interleukin 1 beta (Il-1b), Interleukin 6 (Il-6), Interleukin 8 (Il-8) Interleukin 10 (Il-10), Interleukin 12p70 (Il12p70), Interferon-gamma (IFN- ⁇ ) and Tumor Necrosis Factor alpha (TNF ⁇ ) are quantitated and reported as picograms per millilitre (pg/mL).
- Serum is analysed using the Meso Scale Discovery mouse IL-10 and TNF- ⁇ Ultrasensitive kit.
- mice Long term feeding of mice (115 days) with Bif. AH1714 is associated with an increase in the anti-inflammatory cytokine IL-10 from stimulated ex vivo PBMCs, compared to placebo group (fed PBS) for healthy mice (See FIG. 4 (B)) or in a Sepsis/Inflammation model (mice challenged with LPS; See FIG. 4 (C)).
- mice Long term feeding of mice (115 days) with Bif. AH1714 is associated with a decrease in the pro-inflammatory and Th1 cytokines TNF- ⁇ 0 IFN- ⁇ and IL-12 (p70 sub unit) from stimulated ex vivo PBMCs, compared to placebo group (fed PBS) in a Sepsis/Inflammation model (mice challenged with LPS; See FIG. 5 (B), FIG. 6 (B) and FIG. 7 (B)).
- mice Long term feeding mice (115 days) with Bif. AH1714 is associated with an increase in the serum levels of anti-inflammatory cytokine IL-10 and a decrease in the pro-inflammatory and Th1 cytokine TNF- ⁇ , compared to placebo group (fed PBS) in a Sepsis/Inflammation model (mice challenged with LPS; See FIG. 8 (A & B)).
- Example 5 Bif 1714 has Immunomodulatory Activity when Co-Incubated with Human Immune System Cells In Vitro, Different to that of Bif. AH35624
- Bifidobacterium longum infantis strain UCC35624 (B624), two independent culture batches (1 & 2) and Bifidobacterium longum strain 1714 is assayed using a PBMC cytokine induction assay.
- Bacteria are prepared for co-culture experiments in the following formats. Bacteria are grown under anaerobic conditions at 37° C. in Difco MRS Media and harvested just after entering into stationary phase. Freeze dried powders are generated for each of these bacteria and stored at ⁇ 80° C. in pre-aliquoted 100 mg vials. Immediately prior to their use, one aliquot of each strain is removed from the freezer and allowed to reach room temperature. Each strain is washed 3 times in 10 ml ringers followed by centrifugation. A fresh vial is used on each occasion.
- Direct microscopic counts are performed using a Petroff-Hausser counting chamber as per the manufacturer's instructions and washed cells normalized by cell number before addition to the PBMC assay.
- Bacteria (20 ⁇ l in phosphate buffered saline (PBS)) are added to each well of PBMCs to give the total number of bacteria as indicated for each experiment.
- PBS phosphate buffered saline
- PBMCs Peripheral blood mononuclear cells
- BD Vacutainer CPT tubes BD catalog 362761
- PBMCs are washed and resuspended in Dulbecco's Modified Eagle Medium-GlutamaxTM (Glutamax (Glutamine substitute)+pyruvate+4.5 g/I glucose (Gibco catalog 10569-010) 10% fetal bovine serum (Sigma catalog F4135), and 1% penicillin/streptomycin (Sigma catalog P0781).
- PBMCs are incubated (2 ⁇ 10 5 cells per well) in flat-bottomed 96-well plates and 20 ⁇ l of a bacterial suspension.
- a no-bacteria control also is run. All assays are done in triplicate. After a 2-day incubation at 37° C., the plates were spun at 300 ⁇ g, and the supernatants were removed and stored frozen at ⁇ 80° C. until analysis. PBMCs are co-incubated with bacteria for 48 hours at 37° C./5% CO 2 in an incubator. After the 2 day incubation period, the plates are centrifuged at 300 ⁇ g, and the supernatants removed and stored frozen at ⁇ 80° C. until analysis. Cytokines in the culture supernatants are assayed using a 96-well assay kit from Meso Scale Discovery (Gaithersburg, Md.; catalog K15008B-1).
- Human Interleukin 1 beta (Il-1b), Human Interleukin 6 (Il-6), Human Interleukin 8 (Il-8) Human Interleukin 10 (Il-10), Human Interleukin 12p70 (Il12p70), Human Interferon-gamma (IFN- ⁇ ) and Human Tumor Necrosis Factor alpha (TNF ⁇ ) are quantitated and reported as picograms per millilitre (pg/mL). Each sample is assayed in duplicate.
- Bifidobacterium longum infantis strain UCC35624 (B624), two independent culture batches (1 & 2) and Bifidobacterium longum strain 1714 are assayed for immuno-modulation using a PBMC cytokine induction assay with 1.0E+07 bacteria. Supernatants are assayed for a range of cytokines, including IL-1 ⁇ , -6, -8, -10 and -12, TNF- ⁇ and IFN- ⁇ .
- strain 1714 By comparison with 35624 (both cultures of which gave a similar pattern for all cytokines measured), strain 1714 exhibited a very similar pattern for many of the cytokines measured. Surprisingly however, 1714 gave quite a different pattern for IL-12, IFN ⁇ and IL-6.
- IL-6 Incubation with 1714 induces a significantly lower level of IL-6 compared to 35624 at 1.0 ⁇ 10 7 bacteria per well (See Table 5)
- IL-12 Incubation with 1714 induces a significantly lower level of IL-12 compared to 35624 at 1.0 ⁇ 10 7 bacteria per well (See Table 6)
- INF- ⁇ Incubation with 1714 induces a significantly lower level of INF- ⁇ compared to 35624 at 1.0 ⁇ 10 7 bacteria per well (See Table 6)
- IL-6 is a cytokine strongly implicated in the pathology of IBS.
- L-6 is relevant to many disease processes such as diabetes, atherosclerosis, depression, Alzheimer's Disease, systemic lupus erythematosus and rheumatoid arthritis. Hence there is an interest in developing anti-IL-6 agents as therapy against many of these diseases.
- NFkBlux transgenic mice on a C57BL/6J-CBA/J background are obtained from Charles River Laboratories (Wilmington, USA) and bred in-house. Mice are housed under barrier maintained conditions.
- mice are administered Bif. AH1714, as a freeze-dried powder reconstituted in water at approximately 1 ⁇ 10 9 colony forming units/day/animal, or a placebo control. Mice consume the commensal micro-organism in their drinking water ad libitum for 20 days prior to LPS challenge.
- NFkB activity is measured following the administration of the substrate luciferin and imaged using the Xenogen IVIS 100. Baseline NFkB activity is measured prior to challenge with a single 0.5 mg/kg dose of LPS. After 3 hours all animals are then reimaged. Whole body NFkB activity is assessed by subtracting baseline readings.
- Bif. AH1714 reduces systemic LPS-induced NFkB activity in an in-vivo murine Sepsis/Inflammation model as demonstrated by a decreased NFkB activity in spleens isolated 3 hours post challenge (See FIG. 9 ) and from whole animal imaging 1.5 hours post challenge (See FIG. 10 ) from 1714-fed animals compared to Placebo-fed animals. These results demonstrate that feeding with Bif. 1714 is associated with a decreased level of systemic inflammation associated with the transcription factor NFkB.
- Example 7-1714 Exhibits Positive Benefits in Animal Models of Depression and Anxiety
- Depression and anxiety are the most common psychiatric disorders with a high prevalence rate in the community.
- Anxiety disorders are usually subdivided into panic disorder, generalised anxiety disorder, post-traumatic disorder and obsessive compulsive disorder.
- Modern antidepressants such as the selective serotonin reuptake inhibitors (e.g. fluoxetine) and selective noradrenergic and serotinergic reuptake inhibitors (e.g. venlafaxine) are widely used to treat these disorders.
- the treatments are not always effective and are not acceptable to patients.
- probiotics might be effective in such conditions is suggested by previous data indicating that the probiotic Bifidobacterium Infantis reduces the stress hormone corticosterone in rodents (9).
- mice demonstrate several escape-oriented behaviours interspersed with temporally increasing bouts of immobility.
- a 6-minute test session is employed which is videotaped. Videotapes are subsequently scored by a highly trained observer who is unaware of the treatment. The parameter recorded is the number of seconds spent immobile.
- AH1714 gives a positive result suggesting possible antidepressant activity.
- AH1714 induced lower immobility time than the vehicle (Veh) which suggests lower depression-like behaviours in 1714-fed animals. This is similar to the impact of conventional antidepressants such as Lexapro®.
- AH1714 in animal models of depression and anxiety behaves in a similar way to a conventional antidepressant.
- the impact observed is similar to that reported in the literature for antidepressants such as SSRIs.
- AH1714 may be of benefit in the treatment of the psychiatric syndromes of depression and anxiety.
- Example 8 1714 Exhibits Positive Benefits on Inflammatory Markers in Diet-Induced Obesity
- gut microbiota may be involved in the development of obesity in the regulation of energy homeostasis, in insulin resistance, non-alcoholic fatty liver disease and in energy, lipid and amino acid metabolism (reviewed by Ley et al., 2009)
- the Diet-Induced Obesity (DID) mouse model was chosen as the most appropriate mouse model for assessing the impact of selected probiotic candidates on obesity and metabolic health and to look at the relationship between obesity and inflammatory markers.
- the DIO mouse model refers to healthy mice fed a high-fat diet to induce obesity over time.
- mice Seven-week old male C57BL/J6 mice were fed a low-fat diet (10% calories from fat; Research Diets, New Jersey; #D12450B), a high-fat diet (DID; 45% calories from fat; Research Diets, New Jersey; #D12451) or a high-fat diet with AH1714 (1 ⁇ 10 9 cfu/day) in drinking water for 14 weeks. All mice were housed in groups of 5 and fresh probiotic aliquots were administered daily. Body weight and food intake were assessed weekly. At the end of 14 weeks the mice were sacrificed and internal organs were removed, weighed and stored at ⁇ 80° C. The spleens removed and splenocyte cytokine assays were carried out as in example 4.
- DID mice gained significantly more fat mass (p ⁇ 0.001) compared to lean controls over the 14-week feeding period. In agreement with previous studies, DIO mice consumed significantly more calories than lean controls, as measured by the cumulative caloric intake over the 14 week period of the study (p ⁇ 0.001).
- LPS stimulated splenocytes innate immunity stimulus
- AH1714 had the effect of lowering the TNF ⁇ and IL-12 cytokine response to LPS ( FIG. 16 ).
- CD3/CD28 stimulated spenocytes adaptive immunity stimulus
- treatment with AH1714 had the effect of lowering the IL6 cytokine response.
- the human immune system plays a significant role in the aetiology and pathology of a vast range of human diseases. Hyper and hypo-immune responsiveness results in, or is a component of, the majority of disease states.
- One family of biological entities, termed cytokines, are particularly important to the control of immune processes. Pertubances of these delicate cytokine networks are being increasingly associated with many diseases.
- diseases include but are not limited to inflammatory disorders, immunodeficiency, inflammatory bowel disease, irritable bowel syndrome, cancer (particularly those of the gastrointestinal and immune systems), diarrhoeal disease, antibiotic associated diarrhoea, paediatric diarrhoea, appendicitis, autoimmune disorders, multiple sclerosis, Alzheimer's disease, rheumatoid arthritis, coeliac disease, diabetes mellitus, organ transplantation, bacterial infections, viral infections, fungal infections, periodontal disease, urogenital disease, sexually transmitted disease, HIV infection, HIV replication, HIV associated diarrhoea, surgical associated trauma, surgical-induced metastatic disease, sepsis, weight loss, anorexia, fever control, cachexia, wound healing, ulcers, gut barrier function, allergy, asthma, respiratory disorders, circulatory disorders, coronary heart disease, anaemia, disorders of the blood coagulation system, renal disease, disorders of the central nervous system, hepatic disease, ischaemia, nutritional disorders, osteop
- cytokine production is specific for each of the probiotic strains examined.
- specific probiotic strains may be selected for normalising an exclusive cytokine imbalance particular for a specific disease type.
- Customisation of disease specific therapies can be accomplished using either a single strain of AH1714 or mutants or variants thereof or a selection of these strains.
- the enteric flora is important to the development and proper function of the intestinal immune system. In the absence of an enteric flora, the intestinal immune system is underdeveloped, as demonstrated in germ free animal models, and certain functional parameters are diminished, such as macrophage phagocytic ability and immunoglobulin production (10). The importance of the gut flora in stimulating non-damaging immune responses is becoming more evident. The increase in incidence and severity of allergies in the western world has been linked with an increase in hygiene and sanitation, concomitant with a decrease in the number and range of infectious challenges encountered by the host. This lack of immune stimulation may allow the host to react to non-pathogenic, but antigenic, agents resulting in allergy or autoimmunity. Deliberate consumption of a series of non-pathogenic immunomodulatory bacteria would provide the host with the necessary and appropriate educational stimuli for proper development and control of immune function.
- Inflammation is the term used to describe the local accumulation of fluid, plasma proteins and white blood cells at a site that has sustained physical damage, infection or where there is an ongoing immune response. Control of the inflammatory response is exerted on a number of levels (11).
- the controlling factors include cytokines, hormones (e.g. hydrocortisone), prostaglandins, reactive intermediates and leukotrienes.
- Cytokines are low molecular weight biologically active proteins that are involved in the generation and control of immunological and inflammatory responses, while also regulating development, tissue repair and haematopoiesis. They provide a means of communication between leukocytes themselves and also with other cell types. Most cytokines are pleiotrophic and express multiple biologically overlapping activities.
- Cytokine cascades and networks control the inflammatory response rather than the action of a particular cytokine on a particular cell type (12). Waning of the inflammatory response results in lower concentrations of the appropriate activating signals and other inflammatory mediators leading to the cessation of the inflammatory response.
- TNF ⁇ is a pivotal proinflammatory cytokine as it initiates a cascade of cytokines and biological effects resulting in the inflammatory state. Therefore, agents which inhibit TNF ⁇ are currently being used for the treatment of inflammatory diseases, e.g. infliximab.
- IBD inflammatory bowel disease
- Current therapies for treating IBD are aimed at reducing the levels of these pro-inflammatory cytokines, including IL-8 and TNF ⁇ .
- Such therapies may also play a significant role in the treatment of systemic inflammatory diseases such as rheumatoid arthritis.
- strains of the present invention may have potential application in the treatment of a range of inflammatory diseases, particularly if used in combination with other anti-inflammatory therapies, such as non-steroid anti-inflammatory drugs (NSAIDs) or Infliximab.
- NSAIDs non-steroid anti-inflammatory drugs
- Infliximab Infliximab
- the inflammatory response may have significant roles to play in the above mechanisms, thus contributing to the decline of the host and progression of the tumour.
- intestinal bacteria can produce, from dietary compounds, substances with genotoxic, carcinogenic and tumour-promoting activity and gut bacteria can activate pro-carcinogens to DNA reactive agents (15).
- species of Bifidobacterium have low activities of xenobiotic metabolizing enzymes compared to other populations within the gut such as bacteroides, eubacteria and clostridia. Therefore, increasing the number of Bifidobacterium bacteria in the gut could beneficially modify the levels of these enzymes.
- TTFC tetanus toxin fragment C
- probiotic organisms The introduction of probiotic organisms is accomplished by the ingestion of the micro-organism in a suitable carrier. It would be advantageous to provide a medium that would promote the growth of these probiotic strains in the large bowel.
- the addition of one or more oligosaccharides, polysaccharides, or other prebiotics enhances the growth of lactic acid bacteria in the gastrointestinal tract.
- Prebiotics refers to any non-viable food component that is specifically fermented in the colon by indigenous bacteria thought to be of positive value, e.g. bifidobacteria, lactobacilli. Types of prebiotics may include those that contain fructose, xylose, soya, galactose, glucose and mannose.
- the combined administration of a probiotic strain with one or more prebiotic compounds may enhance the growth of the administered probiotic in vivo resulting in a more pronounced health benefit, and is termed synbiotic.
- the probiotic strains may be administered prophylactically or as a method of treatment either on its own or with other probiotic and/or prebiotic materials as described above.
- the bacteria may be used as part of a prophylactic or treatment regime using other active materials such as those used for treating inflammation or other disorders especially those with an immunological involvement.
- Such combinations may be administered in a single formulation or as separate formulations administered at the same or different times and using the same or different routes of administration.
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- Medicinal Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Public Health (AREA)
- Animal Behavior & Ethology (AREA)
- Veterinary Medicine (AREA)
- Pharmacology & Pharmacy (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- General Chemical & Material Sciences (AREA)
- Microbiology (AREA)
- Genetics & Genomics (AREA)
- Zoology (AREA)
- Biotechnology (AREA)
- Wood Science & Technology (AREA)
- Immunology (AREA)
- Diabetes (AREA)
- Biomedical Technology (AREA)
- Virology (AREA)
- Polymers & Plastics (AREA)
- Food Science & Technology (AREA)
- Mycology (AREA)
- Oncology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Communicable Diseases (AREA)
- Hematology (AREA)
- Nutrition Science (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- Physical Education & Sports Medicine (AREA)
- Pulmonology (AREA)
- Dermatology (AREA)
- Rheumatology (AREA)
- Molecular Biology (AREA)
- Obesity (AREA)
Abstract
Description
- This application is a continuation application of prior International Application No. PCT/IE2010/000066 filed on Nov. 11, 2010 (aka 12424& WO) which claims the benefit of U.S. application Ser. No. 12/616,752 (aka 11484M) filed Nov. 11, 2009 and U.S. Provisional Application No. 61/344,030 (aka 12424P&) filed May 11, 2010.
- The invention relates to a Bifidobacterium strain and its use as a probiotic bacteria in particular as an immunomodulatory biotherapeutic agent.
- The defense mechanisms to protect the human gastrointestinal tract from colonization by intestinal bacteria are highly complex and involve both immunological and non-immunological aspects (1). Innate defense mechanisms include the low pH of the stomach, bile salts, peristalsis, mucin layers and anti-microbial compounds such as lysozyme (2). Immunological mechanisms include specialized lymphoid aggregates, underlying M cells, called peyers patches which are distributed throughout the small intestine and colon (3). Luminal antigens presented at these sites result in stimulation of appropriate T and B cell subsets with establishment of cytokine networks and secretion of antibodies into the gastrointestinal tract (4). In addition, antigen presentation may occur via epithelial cells to intraepithelial lymphocytes and to the underlying lamina propria immune cells (5). Therefore, the host invests substantially in immunological defense of the gastrointestinal tract. However, as the gastrointestinal mucosa is the largest surface at which the host interacts with the external environment, specific control mechanisms must be in place to regulate immune responsiveness to the 100 tons of food which is handled by the gastrointestinal tract over an average lifetime. Furthermore, the gut is colonized by over 500 species of bacteria numbering 1011-1012/g in the colon. Thus, these control mechanisms must be capable of distinguishing non-pathogenic adherent bacteria from invasive pathogens, which would cause significant damage to the host. In fact, the intestinal flora contributes to defense of the host by competing with newly ingested potentially pathogenic micro-organisms.
- Bacteria present in the human gastrointestinal tract can promote inflammation. Aberrant immune responses to the indigenous microflora have been implicated in certain disease states, such as inflammatory bowel disease. Antigens associated with the normal flora usually lead to immunological tolerance and failure to achieve this tolerance is a major mechanism of mucosal inflammation (6). Evidence for this breakdown in tolerance includes an increase in antibody levels directed against the gut flora in patients with inflammatory bowel disease (IBD).
- The present invention is directed towards a Bifidobacterium strain which has been shown to have immunomodulatory effects, by modulating cytokine levels or by antagonizing and excluding pro-inflammatory micro-organisms from the gastrointestinal tract.
- The invention provides an isolated strain of Bifidobacterium NCIMB 41676.
- The Bifidobacterium strain may be in the form of viable cells. The Bifidobacterium strain may be in the form of non-viable cells. The Bifidobacterium may be isolated from colonic biopsy tissue from a healthy human subject. The Bifidobacterium strain may be significantly immunomodulatory following oral consumption in humans.
- The invention also provides a formulation which comprises a Bifidobacterium strain as described herein. The formulation may further comprise a probiotic material. The formulation may further comprise a prebiotic material. The formulation may further comprise an ingestable carrier. The ingestable carrier may a pharmaceutically acceptable carrier such as a capsule, tablet or powder. The ingestable carrier may be a food product such as acidified milk, yoghurt, frozen yoghurt, milk powder, milk concentrate, cheese spreads, dressings or beverages. The formulation may further comprise a protein and/or peptide, in particular proteins and/or peptides that are rich in glutamine/glutamate, a lipid, a carbohydrate, a vitamin, mineral and/or trace element. The Bifidobacterium strain may be present in an amount of more than 106 cfu per gram of the formulation. The formulation may further comprise an adjuvant. The formulation may further comprise a bacterial component. The formulation may further comprise a drug entity. The formulation may further comprise a biological compound. The formulation may be used for immunisation and vaccination protocols.
- The invention also provides a Bifidobacterium strain or a formulation as described herein for use in foodstuffs.
- The invention also provides a Bifidobacterium strain or a formulation as described herein for use as a medicament.
- The invention also provides a Bifidobacterium strain or a formulation as described herein for use in the prophylaxis and/or treatment of undesirable inflammatory activity.
- The invention also provides a Bifidobacterium strain or a formulation as described herein for use in the prophylaxis and/or treatment of undesirable gastrointestinal inflammatory activity such as inflammatory bowel disease eg. Crohns disease or ulcerative colitis, irritable bowel syndrome; pouchitis; or post infection colitis.
- The invention also provides a Bifidobacterium strain or a formulation as described herein for use in the prophylaxis and/or treatment of gastrointestinal cancer(s).
- The invention also provides a Bifidobacterium strain or a formulation as described herein for use in the prophylaxis and/or treatment of systemic disease such as rheumatoid arthritis.
- The invention also provides a Bifidobacterium strain or a formulation as described herein for use in the prophylaxis and/or treatment of autoimmune disorders due to undesirable inflammatory activity.
- The invention also provides a Bifidobacterium strain or a formulation as described herein for use in the prophylaxis and/or treatment of cancer due to undesirable inflammatory activity.
- The invention also provides a Bifidobacterium strain or a formulation as described herein for use in the prophylaxis of cancer.
- The invention also provides a Bifidobacterium strain or a formulation as described herein for use in the prophylaxis and/or treatment of diarrhoeal disease due to undesirable inflammatory activity, such as Clostridium difficile associated diarrhoea, Rotavirus associated diarrhoea or post infective diarrhoea or diarrhoeal disease due to an infectious agent, such as E. coli.
- The invention also provides a Bifidobacterium strain or a formulation as described herein for use in the preparation of anti-inflammatory biotherapeutic agents for the prophylaxis and/or treatment of undesirable inflammatory activity.
- Bifidobacterium strains as described herein may be used in the preparation of a panel of biotherapeutic agents for modifying the levels of IL-10.
- The invention also provides a Bifidobacterium strain or a formulation as described herein for use in the prevention and/or treatment of inflammatory disorders, immunodeficiency, inflammatory bowel disease, irritable bowel syndrome, cancer (particularly of the gastrointestinal and immune systems), diarrhoeal disease, antibiotic associated diarrhoea, paediatric diarrhoea, appendicitis, autoimmune disorders, multiple sclerosis, Alzheimer's disease, rheumatoid arthritis, coeliac disease, diabetes mellitus, organ transplantation, bacterial infections, viral infections, fungal infections, periodontal disease, urogenital disease, sexually transmitted disease, HIV infection, HIV replication, HIV associated diarrhoea, surgical associated trauma, surgical-induced metastatic disease, sepsis, weight loss, anorexia, fever control, cachexia, wound healing, ulcers, gut barrier function, allergy, asthma, respiratory disorders, circulatory disorders, coronary heart disease, anaemia, disorders of the blood coagulation system, renal disease, disorders of the central nervous system, hepatic disease, ischaemia, nutritional disorders, osteoporosis, endocrine disorders, epidermal disorders, psoriasis, acne vulgaris, panic disorder, behavioral disorder and/or post traumatic stress disorders.
- The Bifidobacterium strain as described herein may act by antagonising and excluding proinflammatory micro-organisms from the gastrointestinal tract.
- The invention also provides a Bifidobacterium strain or a formulation as described herein for use in the preparation of anti-inflammatory biotherapeutic agents for reducing the levels of pro inflammatory cytokines.
- The Bifidobacterium strain as described herein may be used as an anti-infective probiotic strain.
- The invention also provides a Bifidobacterium strain or a formulation as described herein for use in the prophylaxis and/or treatment of bipolar illness, depression, mood disorders, and/or anxiety disorders.
- The invention also provides a Bifidobacterium strain or a formulation as described herein may be used as a cognative enhancer for the prophylaxis and/or treatment of disorders of the central nervous system such as Alzheimer's disease, schizophrenia and/or mild cognative disorder.
- The invention also provides a Bifidobacterium strain or a formulation as described herein for use in the prophylaxis and/or treatment of obesity related inflammation.
- The invention also provides a Bifidobacterium strain or a formulation as described herein for use in the prophylaxis and/or treatment of obesity related metabolic dysregulation.
- We describe Bifidobacterium strain AH1714 (NCIMB 41676) or mutants or variants thereof. The mutant may be a genetically modified mutant. The variant may be a naturally occurring variant of Bifidobacterium. Also described is a rifampicin resistant variant of strain AH1714. The strain may be a probiotic. It may be in the form of a biologically pure culture.
- We also describe an isolated strain of Bifidobacterium NCNB 41676. The
Bifidobacterium 30 strains may be in the form of viable cells. Alternatively Bifidobacterium strains are in the form of non-viable cells. The general use of probiotic bacteria is in the form of viable cells. However, it can also be extended to non-viable cells such as killed cultures or compositions containing beneficial factors expressed by the probiotic bacteria. This could include thermally killed micro-organisms or micro-organisms killed by exposure to altered pH or subjection to pressure or gamma irradiation. With non-viable cells product preparation is simpler, cells may be incorporated easily into pharmaceuticals and storage requirements are much less limited than viable cells. Lactobacillus casei YIT 9018 offers an example of the effective use of heat killed cells as a method for the treatment and/or prevention of tumour growth as described in U.S. Pat. No. 4,347,240. - The Bifidobacterium strains may be isolated from colonic biopsy tissue from healthy human subjects, the Bifidobacterium strains being significantly immunomodulatory following oral consumption in humans.
- We also describe a formulation which comprises the Bifidobacterium strain as described herein. The formulation may include another probiotic material. The formulation may include a prebiotic material. Preferably the formulation includes an ingestable carrier. The ingestable carrier may be a pharmaceutically acceptable carrier such as a capsule, tablet or powder. Preferably the ingestable carrier is a food product such as acidified milk, yoghurt, frozen yoghurt, milk powder, milk concentrate, cheese spreads, dressings or beverages. The formulation may further comprise a protein and/or peptide, in particular proteins and/or peptides that are rich in glutamine/glutamate, a lipid, a carbohydrate, a vitamin, mineral and/or trace element. The Bifidobacterium strain may be present in the formulation at more than 106 cfu per gram of delivery system. Preferably the formulation includes any one or more of an adjuvant, a bacterial component, a drug entity or a biological compound.
- We also describe a Bifidobacterium strain or a formulation for use as foodstuffs, as a medicament, for use in the prophylaxis and/or treatment of undesirable inflammatory activity, for use in the prophylaxis and/or treatment of undesirable respiratory inflammatory activity such as asthma, for use in the prophylaxis and/or treatment of undesirable gastrointestinal inflammatory activity such as inflammatory bowel disease eg. Crohns disease or ulcerative colitis, irritable bowel syndrome, pouchitis, or post infection colitis, for use in the prophylaxis and/or treatment of gastrointestinal cancer(s), for use in the prophylaxis and/or treatment of systemic disease such as rheumatoid arthritis, for use in the prophylaxis and/or treatment of autoimmune disorders due to undesirable inflammatory activity, for use in the prophylaxis and/or treatment of cancer due to undesirable inflammatory activity, for use in the prophylaxis of cancer, for use in the prophylaxis and/or treatment of diarrhoeal disease due to undesirable inflammatory activity, such as Clostridium difficile associated diarrhoea, Rotavirus associated diarrhoea or post infective diarrhoea, for use in the prophylaxis and/or treatment of diarrhoeal disease due to an infectious agent, such as E. coli.
- We also describe a Bifidobacterium strain or a formulation of the invention for use in the preparation of an anti-inflammatory biotherapeutic agent for the prophylaxis and/or treatment of undesirable inflammatory activity or for use in the preparation of anti-inflammatory biotherapeutic agents for the prophylaxis and/or treatment of undesirable inflammatory activity. The strain may act by antagonising and excluding proinflammatory micro-organisms from the gastrointestinal tract.
- We also describe a Bifidobacterium strain or a formulation for use in the preparation of anti-inflammatory biotherapeutic agents for reducing the levels of pro-inflammatory cytokines.
- The Bifidobacterium strain may be used in the preparation of anti-inflammatory biotherapeutic agents for modifying the levels of IL-10.
- The Bifidobacterium strain may be used as a anti-infective probiotic due to their ability to antagonise the growth of pathogenic species.
- We have found that particular strains of Bifidobacterium elicit immunomodulatory effects in vitro.
- The invention is therefore of major potential therapeutic value in the prophylaxis or treatment of dysregulated immune responses, such as undesirable inflammatory reactions for example asthma.
- 30 Bifidobacterium are commensal microorganisms. They have been isolated from the microbial flora within the human gastrointestinal tract. The immune system within the gastrointestinal tract cannot have a pronounced reaction to members of this flora, as the resulting inflammatory activity would also destroy host cells and tissue function. Therefore, some mechanism(s) exist whereby the immune system can recognize commensal non-pathogenic members of the gastrointestinal flora as being different to pathogenic organisms. This ensures that damage to host tissues is restricted and a defensive barrier is still maintained.
- A deposit of Bifidobacterium longum strain AH1714 was made at the National Collections of Industrial and Marine Bacteria Limited (NCIMB) Ferguson Building, Craibstone Estate, Bucksburn, Aberdeen, AB21 9YA, Scotland, UK on Nov. 5, 2009 and accorded the accession number NCIMB 41676.
- The Bifidobacterium longum may be a genetically modified mutant or it may be a naturally occurring variant thereof.
- Preferably the Bifidobacterium longum may be in the form of viable cells.
- Alternatively the Bifidobacterium longum may be in the form of non-viable cells.
- It will be appreciated that the specific Bifidobacterium strain of the invention may be administered to animals (including humans) in an orally ingestible form in a conventional preparation such as capsules, microcapsules, tablets, granules, powder, troches, pills, suppositories, suspensions and syrups. Suitable formulations may be prepared by methods commonly employed using conventional organic and inorganic additives. The amount of active ingredient in the medical composition may be at a level that will exercise the desired therapeutic effect.
- The formulation may also include a bacterial component, a drug entity or a biological compound.
- In addition a vaccine comprising the strains of the invention may be prepared using any suitable known method and may include a pharmaceutically acceptable carrier or adjuvant.
- Throughout the specification the terms mutant, variant and genetically modified mutant include a strain of Bifidobacteria whose genetic and/or phenotypic properties are altered compared to the parent strain. Naturally occurring variant of Bifidobacterium longum includes the spontaneous alterations of targeted properties selectively isolated. Deliberate alteration of parent strain properties is accomplished by conventional (in vitro) genetic manipulation technologies, such as gene disruption, conjugative transfer, etc. Genetic modification includes introduction of exogenous and/or endogenous DNA sequences into the genome of a Bifidobacteria strain, for example by insertion into the genome of the bacterial strain by vectors, including plasmid DNA, or bacteriophages.
- Natural or induced mutations include at least single base alterations such as deletion, insertion, transversion or other DNA modifications which may result in alteration of the amino acid sequence encoded by the DNA sequence.
- The terms mutant, variant and genetically modified mutant also include a strain of Bifidobacteria that has undergone genetic alterations that accumulate in a genome at a rate which is consistent in nature for all micro-organisms and/or genetic alterations which occur through spontaneous mutation and/or acquisition of genes and/or loss of genes which is not achieved by deliberate (in vitro) manipulation of the genome but is achieved through the natural selection of variants and/or mutants that provide a selective advantage to support the survival of the bacterium when exposed to environmental pressures such as antibiotics. A mutant can be created by the deliberate (in vitro) insertion of specific genes into the genome which do not fundamentally alter the biochemical functionality of the organism but whose products can be used for identification or selection of the bacterium, for example antibiotic resistance.
- A person skilled in the art would appreciate that mutant or variant strains of Bifidobacteria can be identified by DNA sequence homology analysis with the parent strain. Strains of Bifidobacteria having a close sequence identity with the parent strain are considered to be mutant or variant strains. A Bifidobacteria strain with a sequence identity (homology) of 96% or more, such as 97% or more, or 98% or more, or 99% or more with the parent DNA sequence may be considered to be a mutant or variant. Sequence homology may be determined using on-line homology algorithm “BLAST” program, publicly available at http://www.ncbi.nlm.nih.gov/BLAST/.
- Mutants of the parent strain also include derived Bifidobacteria strains having at least 85% sequence homology, such as at least 90% sequence homology, or at least 95% sequence homology to the 16s-23s intergenic spacer polynucleotide sequence of the parent strain. These mutants may further comprise DNA mutations in other DNA sequences in the bacterial genome.
- The invention will be more clearly understood from the following description thereof given by way of example only with reference to the accompanying drawings in which:—
-
FIG. 1 is a graph illustrating transit of B. longum AH1714 through the gastrointestinal tract; -
FIG. 2 is a photograph of B. longum AH1714 grown on a Congo Red Agar plate; -
FIG. 3 is a bar chart illustrating the IL-10:IL-12p70 ratio for PBMCs stimulated with Bifidobacterium longum strain 1714 (Bifidobacterium 1714); -
FIG. 4 is a bar chart showing the induction profile of IL-10 in splenocytes isolated from both 1714 and PBS fed mice with and without invivo LPS challenge 1 mg/kg. In vitro cells are either unstimulated (A), stimulated with LPS (B) or stimulated with antiCD3/CD28 (C). Data is shown as Average & SEM; -
FIG. 5 is a bar chart showing the induction profile of TNF-α in splenocytes isolated from both 1714 and PBS fed mice with and without invivo LPS challenge 1 mg/kg. In vitro cells are either unstimulated (A) or stimulated with antiCD3/CD28 (B). Data is shown as Average & SEM; -
FIG. 6 is a bar chart showing the induction profile of IFN-γ in splenocytes isolated from both 1714 and PBS fed mice with and without invivo LPS challenge 1 mg/kg. In vitro cells are either unstimulated (A) or stimulated with antiCD3/CD28 (B). Data is shown as Average & SEM. -
FIG. 7 is a bar chart showing the induction profile of IL-12p70 in splenocytes isolated from both 1714 and PBS fed mice with and without invivo LPS challenge 1 mg/kg. In vitro cells are either unstimulated (A) or stimulated with antiCD3/CD28 (B). Data is shown as Average & SEM; -
FIG. 8 is a bar chart showing the induction profile of TNF-α (A) and IL-10 (B) in serum sampled from both 1714 and PBS fed mice post 2H in vivo challenge withLPS 1 mg/kg. Data is shown as Average & SEM; -
FIG. 9 is a bar chart showing NFkB activity (Photons/second) fromisolated spleen 3 hours post challenge with a single 0.5 mg/kg dose of LPS, from Placebo and 1714-fed animals (** designates p<0.01); -
FIG. 10 is a bar chart (A) showing NFkB activity (Photons/second) from whole body imaging 1.5 hours post challenge with a single 0.5 mg/kg dose of LPS, from Placebo and 1714-fed animals ((B) and (C) are whole body representative images in black and white and colour; -
FIG. 11 is a bar chart representing the time of immobility displayed by the mice over a 6-min test; -
FIG. 12 is a line graph representing the freezing percentage in response to the fearful stimulus (context) for day 1 (acquisition), day 2 (memory/extinction) and day 3 (extinction); -
FIG. 13 is a line graph representing the freezing percentage in response to the fearful stimulus (cue) for day 1 (acquisition), day 2 (memory/extinction) and day 3 (extinction); -
FIG. 14 is a bar chart representing the number of marbles buried by the mice over a 30-min session; -
FIG. 15 is a bar chart representing the body temperature variation (ΔT) mice displayed following handling; and -
FIG. 16 is a bar chart showing the changes in cytokine levels in stimulated splenocytes from the Diet Induced Mouse model. - A deposit of Bifidobacterium longum strain AH1714 was made at the National Collections of Industrial and Marine Bacteria Limited (NCIMB) Ferguson Building, Craibstone Estate, Bucksburn, Aberdeen, AB21 9YA, Scotland, UK on Nov. 5, 2009 and accorded the accession number NCIMB 41676.
- A deposit of Bifidobacterium longum strain UCC35624 was made at the National Collections of Industrial and Marine Bacteria Limited (NCIMB) Ferguson Building, Craibstone Estate, Bucksburn, Aberdeen, AB21 9YA, Scotland, UK on Jan. 13, 1999 and accorded the accession number NCIMB 41003.
- The following examples further describe and demonstrate embodiments within the scope of the invention. The examples are given solely for the purpose of illustration and are not to be construed as limitations of the present invention, as many variations thereof are possible without departing from the spirit and scope of the invention.
- Bifidobacterium longum strain AH1714 was isolated from colonic biopsy tissue from healthy human subjects.
- Sections of the large of the human G.I.T, obtained during colorectal scoping, were screened for probiotic bacterial strains. Musocal tissue from the human gastrointestinal tract was transferred to a collection tube containing Phosphate Buffered Saline (PBS), supplemented with 0.05% cysteine-HCl). Triton X-100 (0.05%) was added to release the adherent microorganisms from the tissue sample. Tissue samples were then incubated for 10 min. The samples were vortexed vigorously and adherent Lactobacilli and Bifidobacteria isolated from the gastrointestinal tissue by plating on selective agar (De Man, Rogosa and Sharpe (MRS) agar+Vancomycin and Wilkins-Chalgren Agar+Mupirocin, respectively). Isolated colonies were picked from the plates and re-streaked three times to ensure purity. Microscope examination, Gram staining, Catalase testing, Fructose-6-Phosphate Phosphoketolase assessment were used to determine presumptive Bifidobacteria species and isolates were stocked in 40% glycerol and stored at −20° and −80° C. 16S intergenic spacer region sequencing were used to confirm the identity of the newly isolated strains.
- Following isolation of a pure bifidobacteria strain, assigned the designation AH1714, microbiological characteristics were assessed and are summarized in Table 1 below. AH1714 is a gram positive, catalase negative pleomorphic shaped bacterium which is Fructose-6-Phosphate Phosphoketolase positive, confirming its identity as a bifidobacterium.
-
TABLE 1 Physiochemical characteristics of B. longum AH1714 Strain Characteristics B. longum AH1714 Gram Stain + Catalase − Motility − F6PPK* + - 16s-23s intergenic spacer (IGS) sequencing was performed to identify the species of Bifidobacteria isolated. Briefly, DNA was isolated from AH1714 using 100 μl of Extraction Solution and 25 μl of Tissue Preparation solution (Sigma, XNAT2 Kit). The samples were incubated for 5 minutes at room temperature followed by 2 hrs at 95° C. and then 100 μl of Neutralization Solution (Sigma, XNAT2 kit) was added. Genomic DNA solution was quantified using a Nanodrop spectrophotometer and stored at 4° C. PCR was performed using the IGS primers. The primer pairs used were
IGS R 5′-CTGGTGCCAAGGCATCCA-3′ (SEQ ID No. 4) andIGS L 5′-GCTGGATCACCTCCTTTCT-3′ (SEQ ID No. 3). The cycling conditions were 94° C. for 4 min (1 cycle), 94° C. for 45 sec, 53° C. for 45 sec, 72° C. for 45 sec (28 cycles). The PCR reaction contained 2 μl (100 ng) of DNA, PCR mix (Sigma, Red Taq), 0.025 nM IGS L and R primer (MWG Biotech, Germany). The PCR reactions were performed on a Biotherma thermocycler. The PCR products (10 μl) were ran alongside a molecular weight marker (100 bp Ladder, Roche) on a 2% agarose EtBr stained gel in TAE, to determine the IGS profile. PCR products of Bifidobacterium (single band) were purified using the Promega Wizard PCR purification kit. The purified PCR products were sequenced using the primer sequences (above) for the intergenic spacer region. Sequence data was then searched against the NCBI nucleotide database to determine the identity of the strain by nucleotide homology. The resultant DNA sequence data was subjected to the NCBI standard nucleotide-to-nucleotide homology BLAST search engine (http://www.ncbi.nlm.nih.gov/BLAST/). The nearest match to the sequence was identified and then the sequences were aligned for comparison using DNASTAR MegAlign software. The sequences (SEQ ID NO. 1 [IGS forward sequence] and SEQ ID NO. 2 [IGS reverse sequence]) obtained can be viewed in the sequence listing. Searching the NCIMB database revealed that AH1714 has a unique IGS (SEQ ID NO. 1 [forward sequence] and SEQ ID NO. 2 [reverse sequence]) sequence with its closest sequence homology to a Bifidobacterium longum. - In order to develop a barcode PCR profile for AH1714, PCR was performed using BOX primers (8). The cycling conditions were 94° C. for 7 min (1 cycle); 94° C. for 1 minute, 53° C. for 45 secs, 65° C. for 8 minutes, (30 cycles) and 65° C. for 16 minutes. The PCR reaction contained 50 ng of DNA, PCR mix (Sigma, Red Taq) and 0.03 nM BOXA1R primer (5′-CTACGGCAAGGCGACGCTGACG-3′) (SEQ ID NO. 5) (MWG Biotech, Germany). The PCR reactions were performed on a Biotherma thermocycler. The PCR products were run on a 3% agarose gel alongside a molecular weight marker (Roche, 100 bp ladder) and imaged.
- The antibiotic sensitivity profiles for B. longum AH1714 was determined using the ‘disc susceptibility’ assay. The cultures were grown up in the appropriate broth medium for 48 h and spread-plated (100 μl) onto agar media. Discs containing known concentrations of the antibiotics were placed onto the agar. Strains were examined for antibiotic sensitivity after 1-2 days incubation at 37° C. under anaerobic conditions.
-
TABLE 2 antibiotic resistance Antibiotic Group AH1714 Penicillin G β-lactam antibiotic S Ampicillin β-lactam antibiotic S Methicillin β-lactam antibiotic M Streptomycin Aminoglycoside antibiotic R Gentamicin Aminoglycoside antibiotic M Vancomycin Glycopeptide antibiotic S Nalidixic Acid Synthetic quinolone antibiotic R Novobiocin Aminocoumarin antibiotic S Tetracycline Polyketide antibiotic S Sulphamethoxazole Sulfonamide antibiotic R Trimethoprim\ Sulphamethoxazole Sulfonamide antibiotic R Trimethoprim R Rifampicin Rifamycin antibiotic S Chloramphenicol S Metronidazole Nitroimidazole antibiotics M Mupirocin R R = Resistant (Zones size ≤14 mm) M = Moderately sensitive (Zone size 15-19 mm) S = Sensitive (Zone size ≥20 mm) - Intestinal Transit
- To determine whether Bifidobacterium longum AH1714 could survive at low pH values equivalent to those found in the stomach, bacterial cells were harvested from fresh overnight cultures, washed twice in phosphate buffer (pH 6.5) and resuspended in TPY broth adjusted to pH 2.5 (with 1M HCl). Cells were incubated at 37° C. and survival measured at intervals of 5, 30, 60 and 120 minutes using the plate count method. AH1714 survived well for 5 minutes at pH 2.5 while no viable cells were recovered after 30 minutes.
- Upon exiting the stomach, putative probiotics are exposed to bile salts in the small intestine. In order to determine the ability of B. longum AH1714 to survive exposure to bile, cultures were streaked on TPY agar plates supplemented with 0.3% (w/v), 0.5%, 1%, 2%, 5%, 7.5% or 10% porcine bile. B. longum AH1714 growth was observed on plates containing up to 0.5% bile.
-
TABLE 3 Growth of AH1714 in the presence of porcine bile (duplicate results) % (w/v) Porcine bile Strain 0.0 0.3 0.5 1.0 2.0 5.0 7.5 10.0 AH 1714+++ ++ + − − − − − +++ = very good growth~100% ++ = good growth~66% + = poor growth~33% − = no growth~0% - In a germ-free murine model, the ability of B. longum AH1714 to transit the gastrointestinal tract was assessed. Mice consumed 1×109 AH1714 daily and faecal pellets were examined for the presence of the fed micro-organism. Detection of AH1714 was facilitated by isolating a spontaneous rifampicin resistant variant of the bifidobacteria—incorporation of rifampicin in the RCA+cysteine plates used to assess transit ensured that only the fed rifampicin resistant bifiobacteria was cultured. Faecal samples were collected daily and B. longum AH1714 transit through the gastrointestinal tract was confirmed (See
FIG. 1 ) - To assess the antimicrobial activities of B. longum AH1714 against indicator cultures and to determine if the antimicrobial activity was due to acid production, AH1714 was grown overnight in MRS (supplemented with 0.05% cysteine-HCl). 2 μl of AH1714 culture was spotted onto the agar and incubated for 24 h. The indicator organisms were grown in TSB (E. coli and Salmonella typhimurium), Brucella broth (Campylobacter jejuni) and Reinforced Clostridia Media (RCM, Clostridium difficile). The indicator lawn was prepared by inoculating a molten overlay with 2% 0.5 (v/v) of the overnight indicator culture, which was poured onto the surface of the spotted probiotic cultures following overnight growth on the agar plates. Plates were incubated at 37° C. under suitable conditions for the indicator strain and the growth recorded after 24-48 hr. Zones of clearing greater than 1 mm diameter were considered sensitive to the probiotic strain. This assay was also performed on media supplemented with 2% 3-glycerophosphate as a buffering agent to limit antagonistic activity due to acid production.
-
TABLE 4 antimicrobial activity of AH1714 Zone of inhibition (mm) Indicator strain Non-Buffered Buffered Campylobacter jejuni 9 9 Clostridium petfringens 20 10 Salmonella typhimurium 19 11 E. coli O157:H7 16 11 - In order to track transit of AH1714 in faecal samples, a spontaneous rifampicin-resistant variant (rif+) was isolated as follows: a fresh broth culture of AH1714 was spread-plated (100 μL) onto MRS+rifampicin+cysteine with the lowest concentration of rifampicin (range was 0.1%, 0.08%, 0.06%, 0.04%, 0.02% and 0.002%). Plate media without rifampicin was included as a positive control. Both sets of plates were incubated anaerobically at 37° C. (48 hours). The removed plates were assessed for purity before picking one colony from the rifampicin supplemented agar plate and streaking onto the rifampicin supplemented plate of next highest concentration. In addition, a colony was streaked from the MRS agar plate onto a fresh MRS agar plate and both sets of plates incubated anaerobically at 37° C. (48 hours). This process was repeated for the full range of rifampicin supplemented plates. A single colony from a fully grown culture on a 50 μg/mL rifampicin supplemented MRS agar plate was used to inoculate into 20 ml MRS broth and the resultant culture used for subsequent stocking. The identity of the variant was confirmed by microscopic assessment, IGS sequence analysis and by specific PCR analysis.
- A Congo red agar screen was used to phenotypically screen for EPS expressing bacterial strains. Briefly, 10 ml Modified Rogosa broth media (+0.05% cysteine) was inoculated aseptically with a freshly grown colony of the bacterial strain and incubated anaerobically at 37° C. until turbid (about 16 to about 24 hours). The broth cultures were aseptically streaked onto Congo Red Agar plates and incubated anaerobically at 37° C. for 48 hours. It is believed that EPS produced as a by-product of the growth and/or metabolism of certain strains prevents the uptake of the Congo red stain resulting in a cream/white colony morphology. Stains that produce less EPS take up the Congo red stain easily, resulting in a pink/red colony morphology. Strains that do not produce an EPS stain red and look almost transparent in the red agar background.
- Referring to
FIG. 2 the colony morphology for B. longum AH1714 is convex, mucoid, bright white colonies - Peripheral blood mononuclear cells (PBMCs) were isolated from healthy human peripheral blood using BD Vacutainer CPT tubes (BD catalog 362761), as per the manufacturer's instructions. PBMCs were washed and resuspended in Dulbecco's Modified Eagle Medium-Glutamax™ (Glutamax (Glutamine substitute)+pyruvate+4.5 g/l glucose (Gibco catalog 10569-010) 10% fetal bovine serum (Sigma catalog F4135), and 1% penicillin/streptomycin (Sigma catalog P0781). PBMCs were incubated (2×105 cells per well) in flat-bottomed 96-well plates and 20 μL of a bacterial suspension (at a concentration of 1×107 CFU/mL) was added. PBMCs were co-incubated with bacteria for 48 hours at 37° C./5% CO2 in an incubator. After the 2 day incubation period, the plates were centrifuged at 300×g, and the supernatants were removed and stored frozen at −80° C. until analysis. Interleukin-10 (IL-10) and Interleukin-12p70 (IL-12p70) levels in the culture supernatants were quantified using a 96-well assay kit from Meso Scale Discovery (Gaithersburg, Md.; catalog K15008B-1)
- Bacteria were prepared for co-culture experiments in two formats. (a) Freshly grown bacteria were grown in Difco MRS media and harvested just after entering into stationary phase. All cells were grown under anaerobic conditions at 37° C. (b) Bacteria were grown under anaerobic conditions at 37° C. in Difco MRS media and harvested just after entering into stationary phase. Freeze dried powders were generated for each of these bacteria and stored at −80° C. in pre-aliquoted 100 mg vials. Immediately prior to their use, one aliquot of each strain was removed from the freezer and allowed to reach room temperature. Each strain was washed 3 times in 10 ml ringers followed by centrifugation. A fresh vial was used on each occasion. Growth curves (OD vs number of live cells) were constructed for each growth condition, and washed cells were normalized by cell number before addition to the PBMCs. A no-bacteria control was also included in all experiments. All assays were done in triplicate. The results are presented in
FIG. 3 . - The control of inflammatory diseases is exerted at a number of levels. The controlling factors include hormones, prostaglandins, reactive oxygen and nitrogen intermediates, leukotrienes and cytokines. Cytokines are low molecular weight biologically active proteins that are involved in the generation and control of immunological and inflammatory responses. A number of cell types produce these cytokines, with neutrophils, monocytes and lymphocytes being the major sources during inflammatory reactions due to their large numbers at the injured site.
- Multiple mechanisms exist by which cytokines generated at inflammatory sites influence the inflammatory response. Chemotaxis stimulates homing of inflammatory cells to the injured site, whilst certain cytokines promote infiltration of cells into tissue. Cytokines released within the injured tissue result in activation of the inflammatory infiltrate. Most cytokines are pleiotropic and express multiple biologically overlapping activities. As uncontrolled inflammatory responses can result in diseases such as IBD, it is reasonable to expect that cytokine production has gone astray in individuals affected with these diseases.
- Interleukin-10 (IL-10) is an anti-inflammatory cytokine which is produced by many cell types including monocytes, macrophages, dendritic cells, mast cells and lymphocytes (in particular T regulatory cells). IL-10 down-regulates the expression of pro-inflammatory Th1 cytokines, MHC class II antigens, and co-stimulatory molecules on antigen presenting cells. It also enhances B cell survival, proliferation, and antibody production. This cytokine can block NF-κB activity, and is involved in the regulation of the JAK-STAT signaling pathway. Murine knock-out studies have demonstrated the essential role for IL-10 in immunoregulation as IL-10KO mice develop severe colitis. In addition, bacteria which are potent inducers of 1-10 have been shown to promote T regulatory cell differentiation in vivo thus contributing to immunological homeostasis (7; 8). Interleukin-12 (IL-12) is a pro-inflammatory cytokine associated with polarisation of Th1 effector T cell responses and stimulates the production of other pro-inflammatory Th1 cytokines, such as interferon-gamma (IFN-γ) and tumor necrosis factor-alpha (TNF-α), from T and natural killer (NK) cells. High levels of IL-12 expression is associated with autoimmunity. Administration of IL-12 to people suffering from autoimmune diseases was shown to worsen disease symptoms. In contrast, IL-12 knock-out mice or treatment of mice with IL-12 neutralising antibodies ameliorated the disease.
- Cytokine cascades and networks control the inflammatory response, rather than the action of a particular cytokine on a particular cell type. The relative levels of expression, or balance, of two cytokines (such as IL-10 and IL-12) is more informative than the expression of a single cytokine. In these studies, we stimulated human PBMCs with a range of different bacterial strains. All strains induced IL-10 and all strains induced IL-12. However, examination of the ratio between IL-10 and IL-12 induction revealed that some bacterial strains induced a higher ratio (i.e. more IL-10 with less 1-12) compared to other strains. This is a meaningful observation as it is the balance between each of these opposing signals that ultimately determines the immunological outcome. It is anticipated that a high IL-10:IL-12 ratio would promote an anti-inflammatory response associated with appropriate immunoregulatory activity while a low IL-10:IL-12 ratio would contribute to Th1 polarisation of the immune response. Thus, the PBMC IL-10:IL-12 ratio is a important selection criterion for identification of bacterial strains with immunoregulatory properties.
- Materials & Methods:
- Female Balb/c mice @ 6-8 weeks of age are sourced from Harlan UK and housed in individually ventilated cages and provided ad libitum access to sterile standard mouse chow and water.
- Mice of similar weight are randomised into 2 groups and administered PBS (carrier control n=9), Bifidobacterium longum strain AH1714 (n=17) via oral gavage on a daily basis for 115 days. Following the period of administration blood is sampled from 10 AH1714 mice and 6 carrier controls were challenged with 1 mg/kg LPS (Sigma, L4391) via intra peritoneal injection. Following the period of administration blood is sampled from 6 AH1714 mice and 4 carrier controls, serum is extracted and preserved for cytokine measurements. Spleens are also removed and single cell suspensions cultured in vitro. Cytokines are measured in cell supernatants following 48 hours culturing.
- A further 10 AH1714 mice and 6 control mice are administered a single dose of LPS @ 1 mg/kg via intraperitoneal injection. After 2 hours blood is sampled and the mice were culled. Serum and splenocyte cells were treated and analysed as previously described.
- Splenocytes are isolated from spleens and incubated for 48 hours at 37° C. (in the presence of penicillin and streptomycin) with control media, LPS, or antiCD3/CD28. Cytokines in the culture supernatants are assayed using a 96-well assay kit from Meso Scale Discovery (Gaithersburg, Md.; catalog K15008B-1).
Interleukin 1 beta (Il-1b), Interleukin 6 (Il-6), Interleukin 8 (Il-8) Interleukin 10 (Il-10), Interleukin 12p70 (Il12p70), Interferon-gamma (IFN-γ) and Tumor Necrosis Factor alpha (TNF□) are quantitated and reported as picograms per millilitre (pg/mL). - Serum is analysed using the Meso Scale Discovery mouse IL-10 and TNF-α Ultrasensitive kit.
- Long term feeding of mice (115 days) with Bif. AH1714 is associated with an increase in the anti-inflammatory cytokine IL-10 from stimulated ex vivo PBMCs, compared to placebo group (fed PBS) for healthy mice (See
FIG. 4 (B)) or in a Sepsis/Inflammation model (mice challenged with LPS; SeeFIG. 4 (C)). - Long term feeding of mice (115 days) with Bif. AH1714 is associated with a decrease in the pro-inflammatory and Th1 cytokines TNF-α□0 IFN-γ and IL-12 (p70 sub unit) from stimulated ex vivo PBMCs, compared to placebo group (fed PBS) in a Sepsis/Inflammation model (mice challenged with LPS; See
FIG. 5 (B),FIG. 6 (B) andFIG. 7 (B)). - Long term feeding mice (115 days) with Bif. AH1714 is associated with an increase in the serum levels of anti-inflammatory cytokine IL-10 and a decrease in the pro-inflammatory and Th1 cytokine TNF-α, compared to placebo group (fed PBS) in a Sepsis/Inflammation model (mice challenged with LPS; See
FIG. 8 (A & B)). - Taken together, these results demonstrate that
Bifidobacterium longum strain 1714 has in-vivo systemic immunomodulatory and anti-inflammatory activity and protects against LPS or TLR-4 mediated inflammatory responses. - Bifidobacterium longum infantis strain UCC35624 (B624), two independent culture batches (1 & 2) and
Bifidobacterium longum strain 1714 is assayed using a PBMC cytokine induction assay. Bacteria are prepared for co-culture experiments in the following formats. Bacteria are grown under anaerobic conditions at 37° C. in Difco MRS Media and harvested just after entering into stationary phase. Freeze dried powders are generated for each of these bacteria and stored at −80° C. in pre-aliquoted 100 mg vials. Immediately prior to their use, one aliquot of each strain is removed from the freezer and allowed to reach room temperature. Each strain is washed 3 times in 10 ml ringers followed by centrifugation. A fresh vial is used on each occasion. - Direct microscopic counts are performed using a Petroff-Hausser counting chamber as per the manufacturer's instructions and washed cells normalized by cell number before addition to the PBMC assay. Bacteria (20 μl in phosphate buffered saline (PBS)) are added to each well of PBMCs to give the total number of bacteria as indicated for each experiment.
- Peripheral blood mononuclear cells (PBMCs) are isolated from healthy human peripheral blood using BD Vacutainer CPT tubes (BD catalog 362761), as per the manufacturer's instructions. PBMCs are washed and resuspended in Dulbecco's Modified Eagle Medium-Glutamax™ (Glutamax (Glutamine substitute)+pyruvate+4.5 g/I glucose (Gibco catalog 10569-010) 10% fetal bovine serum (Sigma catalog F4135), and 1% penicillin/streptomycin (Sigma catalog P0781). PBMCs are incubated (2×105 cells per well) in flat-bottomed 96-well plates and 20 μl of a bacterial suspension. A no-bacteria control also is run. All assays are done in triplicate. After a 2-day incubation at 37° C., the plates were spun at 300×g, and the supernatants were removed and stored frozen at −80° C. until analysis. PBMCs are co-incubated with bacteria for 48 hours at 37° C./5% CO2 in an incubator. After the 2 day incubation period, the plates are centrifuged at 300×g, and the supernatants removed and stored frozen at −80° C. until analysis. Cytokines in the culture supernatants are assayed using a 96-well assay kit from Meso Scale Discovery (Gaithersburg, Md.; catalog K15008B-1).
Human Interleukin 1 beta (Il-1b), Human Interleukin 6 (Il-6), Human Interleukin 8 (Il-8) Human Interleukin 10 (Il-10), Human Interleukin 12p70 (Il12p70), Human Interferon-gamma (IFN-γ) and Human Tumor Necrosis Factor alpha (TNFα) are quantitated and reported as picograms per millilitre (pg/mL). Each sample is assayed in duplicate. - Bifidobacterium longum infantis strain UCC35624 (B624), two independent culture batches (1 & 2) and
Bifidobacterium longum strain 1714 are assayed for immuno-modulation using a PBMC cytokine induction assay with 1.0E+07 bacteria. Supernatants are assayed for a range of cytokines, including IL-1β, -6, -8, -10 and -12, TNF-α□ and IFN-γ. - By comparison with 35624 (both cultures of which gave a similar pattern for all cytokines measured),
strain 1714 exhibited a very similar pattern for many of the cytokines measured. Surprisingly however, 1714 gave quite a different pattern for IL-12, IFNγ and IL-6. - IL-6: Incubation with 1714 induces a significantly lower level of IL-6 compared to 35624 at 1.0×107 bacteria per well (See Table 5)
-
TABLE 5 Strain (1 × 10E7 bacteria) IL-6 (~pg/ml) 35624 28,000 1714 16,000 - IL-12: Incubation with 1714 induces a significantly lower level of IL-12 compared to 35624 at 1.0×107 bacteria per well (See Table 6)
- INF-γ Incubation with 1714 induces a significantly lower level of INF-γ compared to 35624 at 1.0×107 bacteria per well (See Table 6)
-
TABLE 6 Strain (1 × 10E7 IL-12 IL-12 IFN-α IFN-γ bacteria) (~pg/ml) (~pg/ml) (~pg/ml) (~pg/ml) 35624 500 180 5000 1500 1714 (1) 220 115 2400 750 1714 (2) 240 80 2600 400 - Foligne et al19, have demonstrated that lactic acid bacteria strains displaying an in vitro capacity to induce higher levels of the anti-inflammatory cytokine IL-10 and lower levels of the inflammatory cytokine IL-12 offered the best protection in the in vivo colitis model whereas in contrast, strains leading to a low IL-10/IL-12 cytokine ratio could not significantly alleviate colitis symptoms. The in vivo protection observed was strain specific. The cytokine profile obtained for Bif. AH1714 would suggest that this strain has the potential for improved efficacy in the in vivo ulcerative colitis model.
- IL-6 is a cytokine strongly implicated in the pathology of IBS. L-6 is relevant to many disease processes such as diabetes, atherosclerosis, depression, Alzheimer's Disease, systemic lupus erythematosus and rheumatoid arthritis. Hence there is an interest in developing anti-IL-6 agents as therapy against many of these diseases.
- NFkBlux transgenic mice on a C57BL/6J-CBA/J background are obtained from Charles River Laboratories (Wilmington, USA) and bred in-house. Mice are housed under barrier maintained conditions.
- Female animals are administered Bif. AH1714, as a freeze-dried powder reconstituted in water at approximately 1×109 colony forming units/day/animal, or a placebo control. Mice consume the commensal micro-organism in their drinking water ad libitum for 20 days prior to LPS challenge.
- NFkB activity is measured following the administration of the substrate luciferin and imaged using the
Xenogen IVIS 100. Baseline NFkB activity is measured prior to challenge with a single 0.5 mg/kg dose of LPS. After 3 hours all animals are then reimaged. Whole body NFkB activity is assessed by subtracting baseline readings. - All animals are then culled and spleens, livers, small intestine and colon removed and placed in a culture dish for individual imaging.
- Bif. AH1714 reduces systemic LPS-induced NFkB activity in an in-vivo murine Sepsis/Inflammation model as demonstrated by a decreased NFkB activity in spleens isolated 3 hours post challenge (See
FIG. 9 ) and from whole animal imaging 1.5 hours post challenge (SeeFIG. 10 ) from 1714-fed animals compared to Placebo-fed animals. These results demonstrate that feeding with Bif. 1714 is associated with a decreased level of systemic inflammation associated with the transcription factor NFkB. - Depression and anxiety are the most common psychiatric disorders with a high prevalence rate in the community. Anxiety disorders are usually subdivided into panic disorder, generalised anxiety disorder, post-traumatic disorder and obsessive compulsive disorder.
- Modern antidepressants such as the selective serotonin reuptake inhibitors (e.g. fluoxetine) and selective noradrenergic and serotinergic reuptake inhibitors (e.g. venlafaxine) are widely used to treat these disorders. However, the treatments are not always effective and are not acceptable to patients. There is a need to develop alternative strategies. The possibility that probiotics might be effective in such conditions is suggested by previous data indicating that the probiotic Bifidobacterium Infantis reduces the stress hormone corticosterone in rodents (9).
- We here examine the behavioural effects of Bifidobacterium AH1714 in models of stress in mice and compared with a widely used SSRI, namely, escitalopram, which is used to treat both anxiety and depression. Animals are treated with either escitalopram or AH1714 for three weeks.
- A well characterized test for assessing depression-like and antidepressant like activity. Mice are individually suspended by the tail to a horizontal ring-stand bar (distance from floor=30 cm) using adhesive tape (distance from tip of tail=2 cm). Typically, mice demonstrate several escape-oriented behaviours interspersed with temporally increasing bouts of immobility. A 6-minute test session is employed which is videotaped. Videotapes are subsequently scored by a highly trained observer who is unaware of the treatment. The parameter recorded is the number of seconds spent immobile.
- Widely used to assess the cognitive components of anxiety disorders. We use a three day protocol which allows for contextual and cue-associated fear learning to be observed. Following 3 minutes of exploring their environmental context mice receiving 6 pairings of 20 seconds of a specific cue (Tone of 10 KHz, 70 dB combined with apparatus light on) coupled at the end with 2 seconds of a mild electrical footshock (0.4 mA), this is followed by 1 min exposure to the context only. The procedure is repeated for two subsequent days, however, no shock is given and freezing behaviour to the context or cue is observed throughout. The first day allows for assessment of the ability of the intervention to alter the strength of context and cue-induced fear conditioning, whereas the third day allows for extinction of fear learning to be observed. Extinction is the formation of new memories and drugs that facilitate extinction may play a role in the treatment of post-traumatic stress disorder.
- Proposed as model of obsessive compulsive disorder. Animals who are more anxious must engage in active behaviours (defensive marble burying) to avoid anxiogenic stimuli in the light-dark box and elevated mazes. Mice are placed individually in small cages, in which 20 marbles had been equally distributed on top of a 5 cm-deep bed of sawdust, and a wire lid placed on top of the cage. Mice are left undisturbed for 30 min, after which the number of buried marbles (i.e., those more than less than three-quarters covered by sawdust) are counted.
- In the tail suspension test AH1714 gives a positive result suggesting possible antidepressant activity. Referring to
FIG. 11 , AH1714 induced lower immobility time than the vehicle (Veh) which suggests lower depression-like behaviours in 1714-fed animals. This is similar to the impact of conventional antidepressants such as Lexapro®. - In terms of cognition, in the fear conditioning test, test animals treated with AH1714 showed a positive learning effect. Referring to
FIG. 12 , in context (mainly hippocampus and amygdale-dependent memories) fear conditioning tests, 1714 induced higher freezing to the context (Cxt) than the vehicle (Veh) onday 1 andday 2, with the same freezing percentage as the vehicle onday 3, this suggests that 1714 promotes contextual fear learning and memory without impairing extinction, suggesting a positive role in contextual memory of fearful events. Referring toFIG. 13 , in cue (amygdale-dependent) fear conditioning tests, 1714 induced higher freezing to the fearful cue (stimulus) than the vehicle (Veh) onday 1, with the same freezing percentage as the vehicle ondays - Evidence of a possible effect in obsessive compulsive disorder emerges from studies with the marble burying test. Animals treated with AH1714 buried less marbles in the marble burying task which is indicative of lower anxiety in 1714-fed animals and suggests a possible effect in obsessive compulsive disorder (
FIG. 14 ). As in the case of escitalopram, AH1714 induced a lower body temperature increase induced by the stress of being handled (decreased stress induced hypothermia) this suggests lower anxiety in 1714-fed animals (FIG. 24 ). There were no differences between the results with either intervention. - AH1714 in animal models of depression and anxiety behaves in a similar way to a conventional antidepressant. The impact observed is similar to that reported in the literature for antidepressants such as SSRIs.
- Overall, the data indicate that AH1714 may be of benefit in the treatment of the psychiatric syndromes of depression and anxiety.
- In recent years, it has been well established that obesity is associated with a low-grade inflammation that contributes to the development of the pathologies associated with obesity which include
type 2 diabetes mellitus (T2D), cardiovascular disease (CVD), hypertension, hypercholesterolemia, hypertriglyceridemia, and non-alcoholic fatty liver disease (NAFLD). Visceral fat produces a number of inflammatory cytokines and chemokines (such as leptin, tumor necrosis factor-α (TNF-α), macrophage chemo-attractant protein-1 and interleukin-6, among others), whose production can be pathologically dysregulated in the obese state (reviewed by Shoelson et al., 2007). Indeed, while macrophages are thought to contribute in an important manner to insulin resistance, other studies have suggested that harnessing the anti-inflammatory properties of cells with a potentially regulatory phenotype may have therapeutic potential. A recent study suggests that Treg cells reduce the inflammatory state of adipose tissue and, thus, insulin resistance in mice (Feurer et al., 2009). In addition, a seminal body of work has implicated abnormalities of the gut microbiota as a driving force of obesity-related metabolic dysregulation, suggesting that interventions which target gut health will have beneficial health effects in obesity related metabolic derangements. It has been suggested that the gut microbiota may be involved in the development of obesity in the regulation of energy homeostasis, in insulin resistance, non-alcoholic fatty liver disease and in energy, lipid and amino acid metabolism (reviewed by Ley et al., 2009) - The Diet-Induced Obesity (DID) mouse model was chosen as the most appropriate mouse model for assessing the impact of selected probiotic candidates on obesity and metabolic health and to look at the relationship between obesity and inflammatory markers. The DIO mouse model refers to healthy mice fed a high-fat diet to induce obesity over time.
- Seven-week old male C57BL/J6 mice were fed a low-fat diet (10% calories from fat; Research Diets, New Jersey; #D12450B), a high-fat diet (DID; 45% calories from fat; Research Diets, New Jersey; #D12451) or a high-fat diet with AH1714 (1×109 cfu/day) in drinking water for 14 weeks. All mice were housed in groups of 5 and fresh probiotic aliquots were administered daily. Body weight and food intake were assessed weekly. At the end of 14 weeks the mice were sacrificed and internal organs were removed, weighed and stored at −80° C. The spleens removed and splenocyte cytokine assays were carried out as in example 4.
- As expected, DID mice gained significantly more fat mass (p<0.001) compared to lean controls over the 14-week feeding period. In agreement with previous studies, DIO mice consumed significantly more calories than lean controls, as measured by the cumulative caloric intake over the 14 week period of the study (p<0.001). In LPS stimulated splenocytes (innate immunity stimulus) from DID mice, AH1714 had the effect of lowering the TNFα and IL-12 cytokine response to LPS (
FIG. 16 ). In CD3/CD28 stimulated spenocytes (adaptive immunity stimulus), treatment with AH1714 had the effect of lowering the IL6 cytokine response. These results indicate a systemic anti-inflammatory effect in the DIO mouse model consistent with the PBMC data and in vivo mouse model data illustrated in other examples. - The human immune system plays a significant role in the aetiology and pathology of a vast range of human diseases. Hyper and hypo-immune responsiveness results in, or is a component of, the majority of disease states. One family of biological entities, termed cytokines, are particularly important to the control of immune processes. Pertubances of these delicate cytokine networks are being increasingly associated with many diseases. These diseases include but are not limited to inflammatory disorders, immunodeficiency, inflammatory bowel disease, irritable bowel syndrome, cancer (particularly those of the gastrointestinal and immune systems), diarrhoeal disease, antibiotic associated diarrhoea, paediatric diarrhoea, appendicitis, autoimmune disorders, multiple sclerosis, Alzheimer's disease, rheumatoid arthritis, coeliac disease, diabetes mellitus, organ transplantation, bacterial infections, viral infections, fungal infections, periodontal disease, urogenital disease, sexually transmitted disease, HIV infection, HIV replication, HIV associated diarrhoea, surgical associated trauma, surgical-induced metastatic disease, sepsis, weight loss, anorexia, fever control, cachexia, wound healing, ulcers, gut barrier function, allergy, asthma, respiratory disorders, circulatory disorders, coronary heart disease, anaemia, disorders of the blood coagulation system, renal disease, disorders of the central nervous system, hepatic disease, ischaemia, nutritional disorders, osteoporosis, endocrine disorders, epidermal disorders, psoriasis and acne vulgaris.
- The effects on cytokine production are specific for each of the probiotic strains examined. Thus specific probiotic strains may be selected for normalising an exclusive cytokine imbalance particular for a specific disease type. Customisation of disease specific therapies can be accomplished using either a single strain of AH1714 or mutants or variants thereof or a selection of these strains.
- The enteric flora is important to the development and proper function of the intestinal immune system. In the absence of an enteric flora, the intestinal immune system is underdeveloped, as demonstrated in germ free animal models, and certain functional parameters are diminished, such as macrophage phagocytic ability and immunoglobulin production (10). The importance of the gut flora in stimulating non-damaging immune responses is becoming more evident. The increase in incidence and severity of allergies in the western world has been linked with an increase in hygiene and sanitation, concomitant with a decrease in the number and range of infectious challenges encountered by the host. This lack of immune stimulation may allow the host to react to non-pathogenic, but antigenic, agents resulting in allergy or autoimmunity. Deliberate consumption of a series of non-pathogenic immunomodulatory bacteria would provide the host with the necessary and appropriate educational stimuli for proper development and control of immune function.
- Inflammation is the term used to describe the local accumulation of fluid, plasma proteins and white blood cells at a site that has sustained physical damage, infection or where there is an ongoing immune response. Control of the inflammatory response is exerted on a number of levels (11). The controlling factors include cytokines, hormones (e.g. hydrocortisone), prostaglandins, reactive intermediates and leukotrienes. Cytokines are low molecular weight biologically active proteins that are involved in the generation and control of immunological and inflammatory responses, while also regulating development, tissue repair and haematopoiesis. They provide a means of communication between leukocytes themselves and also with other cell types. Most cytokines are pleiotrophic and express multiple biologically overlapping activities. Cytokine cascades and networks control the inflammatory response rather than the action of a particular cytokine on a particular cell type (12). Waning of the inflammatory response results in lower concentrations of the appropriate activating signals and other inflammatory mediators leading to the cessation of the inflammatory response. TNFα is a pivotal proinflammatory cytokine as it initiates a cascade of cytokines and biological effects resulting in the inflammatory state. Therefore, agents which inhibit TNFα are currently being used for the treatment of inflammatory diseases, e.g. infliximab.
- Pro-inflammatory cytokines are thought to play a major role in the pathogenesis of many inflammatory diseases, including inflammatory bowel disease (IBD). Current therapies for treating IBD are aimed at reducing the levels of these pro-inflammatory cytokines, including IL-8 and TNFα. Such therapies may also play a significant role in the treatment of systemic inflammatory diseases such as rheumatoid arthritis.
- The strains of the present invention may have potential application in the treatment of a range of inflammatory diseases, particularly if used in combination with other anti-inflammatory therapies, such as non-steroid anti-inflammatory drugs (NSAIDs) or Infliximab.
- The production of multifunctional cytokines across a wide spectrum of tumour types suggests that significant inflammatory responses are ongoing in patients with cancer. It is currently unclear what protective effect this response has against the growth and development of tumour cells in vivo. However, these inflammatory responses could adversely affect the tumour-bearing host. Complex cytokine interactions are involved in the regulation of cytokine production and cell proliferation within tumour and normal tissues (13, 14). It has long been recognized that weight loss (cachexia) is the single most common cause of death in patients with cancer and initial malnutrition indicates a poor prognosis. For a tumour to grow and spread it must induce the formation of new blood vessels and degrade the extracellular matrix. The inflammatory response may have significant roles to play in the above mechanisms, thus contributing to the decline of the host and progression of the tumour. Due to the anti-inflammatory properties of Bifidobacterium longum infantis these bacterial strains they may reduce the rate of malignant cell transformation. Furthermore, intestinal bacteria can produce, from dietary compounds, substances with genotoxic, carcinogenic and tumour-promoting activity and gut bacteria can activate pro-carcinogens to DNA reactive agents (15). In general, species of Bifidobacterium have low activities of xenobiotic metabolizing enzymes compared to other populations within the gut such as bacteroides, eubacteria and clostridia. Therefore, increasing the number of Bifidobacterium bacteria in the gut could beneficially modify the levels of these enzymes.
- The majority of pathogenic organisms gain entry via mucosal surfaces. Efficient vaccination of these sites protects against invasion by a particular infectious agent. Oral vaccination strategies have concentrated, to date, on the use of attenuated live pathogenic organisms or purified encapsulated antigens (16). Probiotic bacteria, engineered to produce antigens from an infectious agent, in vivo, may provide an attractive alternative as these bacteria are considered to be safe for human consumption (GRAS status).
- Murine studies have demonstrated that consumption of probiotic bacteria expressing foreign antigens can elicit protective immune responses. The gene encoding tetanus toxin fragment C (TTFC) was expressed in Lactococcus lactis and mice were immunized via the oral route. This system was able to induce antibody titers significantly high enough to protect the mice from lethal toxin challenge. In addition to antigen presentation, live bacterial vectors can produce bioactive compounds, such as immunostimulatory cytokines, in vivo. L. lactis secreting bioactive human IL-2 or IL-6 and TTFC induced 10-15 fold higher serum IgG titres in mice immunized intranasally (17). However, with this particular bacterial strain, the total IgA level was not increased by coexpression with these cytokines. Other bacterial strains, such as Streptococcus gordonii, are also being examined for their usefulness as mucosal vaccines. Recombinant S. gordonii colonizing the murine oral and vaginal cavities induced both mucosal and systemic antibody responses to antigens expressed by this bacterial (18). Thus oral immunization using probiotic bacteria as vectors would not only protect the host from infection, but may replace the immunological stimuli that the pathogen would normally elicit thus contributing to the immunological education of the host.
- The introduction of probiotic organisms is accomplished by the ingestion of the micro-organism in a suitable carrier. It would be advantageous to provide a medium that would promote the growth of these probiotic strains in the large bowel. The addition of one or more oligosaccharides, polysaccharides, or other prebiotics enhances the growth of lactic acid bacteria in the gastrointestinal tract. Prebiotics refers to any non-viable food component that is specifically fermented in the colon by indigenous bacteria thought to be of positive value, e.g. bifidobacteria, lactobacilli. Types of prebiotics may include those that contain fructose, xylose, soya, galactose, glucose and mannose. The combined administration of a probiotic strain with one or more prebiotic compounds may enhance the growth of the administered probiotic in vivo resulting in a more pronounced health benefit, and is termed synbiotic.
- It will be appreciated that the probiotic strains may be administered prophylactically or as a method of treatment either on its own or with other probiotic and/or prebiotic materials as described above. In addition, the bacteria may be used as part of a prophylactic or treatment regime using other active materials such as those used for treating inflammation or other disorders especially those with an immunological involvement. Such combinations may be administered in a single formulation or as separate formulations administered at the same or different times and using the same or different routes of administration.
- The dimensions and values disclosed herein are not to be understood as being strictly limited to the exact numerical values recited. Instead, unless otherwise specified, each such dimension is intended to mean both the recited value and a functionally equivalent range surrounding that value. For example, a dimension disclosed as “40 mm” is intended to mean “about 40 mm.”
- Every document cited herein, including any cross referenced or related patent or application, is hereby incorporated herein by reference in its entirety unless expressly excluded or otherwise limited. The citation of any document is not an admission that it is prior art with respect to any invention disclosed or claimed herein or that it alone, or in any combination with any other reference or references, teaches, suggests or discloses any such invention. Further, to the extent that any meaning or definition of a term in this document conflicts with any meaning or definition of the same term in a document incorporated by reference, the meaning or definition assigned to that term in this document shall govern.
- While particular embodiments of the present invention have been illustrated and described, it would be obvious to those skilled in the art that various other changes and modifications can be made without departing from the spirit and scope of the invention. It is therefore intended to cover in the appended claims all such changes and modifications that are within the scope of this invention.
-
- 1. McCracken V. J. and Gaskins H. R. Probiotics and the immune system. In: Probiotics a critical review, Tannock, G W (ed), Horizon Scientific Press, U K. 1999, p. 85-113.
- 2. Savage D. C. Interaction between the host and its microbes. In: Microbial Ecology of the Gut, Clark and Bauchop (eds), Academic Press, London. 1977, p. 277-310.
- 3. Kagnoff M. F. Immunology of the intestinal tract. Gastroenterol. 1993; 105 (5): 1275-80.
- 4. Lamm M. E. Interaction of antigens and antibodies at mucosal surfaces. Ann. Rev. Microbial. 1997; 51: 311-40.
- 5. Raychaudhuri S., Rock K L. Fully mobilizing host defense: building better vaccines. Nat biotechnol., 1998; 16: 1025-31.
- 6. Stallmach A., Strober W, MacDonald T T, Lochs H, Zeitz M. Induction and modulation of gastrointestinal inflammation. Immunol. Today, 1998; 19 (10): 438-41.
- 7. Liam O'Mahony, Louise O'Callaghan, Jane McCarthy, David Shilling, Paul Scully, Shomik Sibartie, Eamon Kavanagh, William O. Kirwan, Henry Paul Redmond, John Kevin Collins, and Fergus Shanahan. Differential cytokine response from dendritic cells to commensal and pathogenic bacteria in different lymphoid compartments in humans. Am J Physiol Gastrointest Liver Physiol 290: G839-G845, 2006.
- 8. O'Mahony C, Scully P, O'Mahony D, Murphy S, O'Brien F, et al. Commensal-Induced Regulatory T Cells Mediate Protection against Pathogen-Stimulated NF-kB Activation. PLoS Pathog 2008, 4(8): e1000112.
- 9. Desbonnet L, Garrett L, Clarke G, Bienenstock J, Dinan T G. The probiotic Bifidobacteria infantis: An assessment of potential antidepressant properties in the rat. J Psychiatr Res. 2008 December; 43(2):164-74.
- 10. Crabbe P. A., H. Bazin, H. Eyssen, and J. F. Heremans. The normal microbial flora as a major stimulus for proliferation of plasma cells synthesizing IgA in the gut. The germ free intestinal tract. Into. Arch. Allergy Appl Immunol, 1968; 34: 362-75.
- 11. Henderson B., Poole, S and Wilson M. 1998. In “Bacteria-Cytokine interactions in health and disease. Portland Press, 79-130.
- 12. Arai K I, Lee F, Miyajima A, Miyatake S, Arai N, Yokota T. Cytokines: coordinators of immune and inflammatory responses. Annu Rev Biochem 1990; 59:783-836.
- 13. McGee D W, Bamberg T, Vitkus S J, McGhee J R. A synergistic relationship between TNF-alpha, IL-1 beta, and TGF-
beta 1 on IL-6 secretion by the IEC-6 intestinal epithelial cell line. Immunology 1995 September; 86(1):6-11. - 14. Wu S, Meeker W A, Wiener J R, Berchuck A, Bast R C Jr, Boyer C M. Transfection of ovarian cancer cells with tumour necrosis factor alpha (TNF-alpha) antisense mRNA abolishes the proliferative response to interleukin-1 (IL-1) but not TNF-alpha. Gynecol Oncol 1994 April; 53(1):59-63.
- 15. Rowland I. R. Toxicology of the colon: role of the intestinal microflora. In: Gibson G. R. (ed). Human colonic bacteria: role in nutrition, physiology and pathology, 1995, pp 155-174. Boca Raton CRC Press.
- 16. Walker, R. I. New strategies for using mucosal vaccination to achieve more effective immunization. Vaccine, 1994; 12: 387-400.
- 17. Steidler L., K. Robinson, L. Chamberlain, K. M Scholfield, E. Remaut, R. W. F. Le Page and J. M. Wells. Mucosal delivery of murine interleukin-2 (IL-2) and IL-6 by recombinant strains of Lactococcus lactis coexpressing antigen and cytokine. Infect. Immun., 1998; 66:3183-9.
- 18. Medaglini D., G. Pozzi, T. P. King and V. A. Fischetti. Mucosal and systemic immune responses to a recombinant protein expressed on the surface of the oral commensal bacterium Streptococcus gordonii after oral colonization. Proc. Natl. Acad. Sci. USA, 1995; 92:6868-72 McCracken V. J. and Gaskins H. R, ‘Probiotics a critical review’, Horizon Scientific Press, UK 1999, p. 278.
- 19. Foligne, B., Nutten, S., Grangette, C., Dennin V., Goudercourt, D., Poiret, S., Dewulf, J., Brassart, D., Mercenier, A., and Pot, B., Correlation between in vitro and in vivo immunomodulatory properties of lactic acid bacteria. World J. Gastroenterol. 2007; 13(2): 236-243.
- 20. Feuerer M, Herrero L, Cipolletta D, Naaz A, Wong J, Nayer A, Lee J, Goldfine A B, Benoist C, Shoelson S, Mathis D. Lean, but not obese, fat is enriched for a unique population of regulatory T cells that affect metabolic parameters. Nature Medicine (2009) 15, 930-939.
- 21. Shoelson S E, Herrero L, Naaz A. Obesity, inflammation, and insulin resistance. Gastroenterology (2007) 132, 2169-2180.
- 22. Ley R E. Obesity and the human microbiome. Curr Opin Gastroenterol. (2010) 1, 5-11.
Claims (21)
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US17/545,148 US20220169976A1 (en) | 2009-11-11 | 2021-12-08 | Probiotic bifidobacterium strain |
Applications Claiming Priority (7)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US12/616,752 US9771624B2 (en) | 2008-11-11 | 2009-11-11 | Bifidobacterium longum |
US34403010P | 2010-05-11 | 2010-05-11 | |
PCT/IE2010/000066 WO2011058535A1 (en) | 2009-11-11 | 2010-11-11 | Probiotic bifidobacterium strain |
US13/468,374 US10144978B2 (en) | 2009-11-11 | 2012-05-10 | Probiotic bifidobacterium strain |
US15/936,661 US10597739B2 (en) | 2009-11-11 | 2018-03-27 | Probiotic bifidobacterium strain |
US16/789,768 US11225641B2 (en) | 2009-11-11 | 2020-02-13 | Probiotic Bifidobacterium strain |
US17/545,148 US20220169976A1 (en) | 2009-11-11 | 2021-12-08 | Probiotic bifidobacterium strain |
Related Parent Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US16/789,768 Continuation US11225641B2 (en) | 2009-11-11 | 2020-02-13 | Probiotic Bifidobacterium strain |
Publications (1)
Publication Number | Publication Date |
---|---|
US20220169976A1 true US20220169976A1 (en) | 2022-06-02 |
Family
ID=43478193
Family Applications (4)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US13/468,374 Active 2031-02-19 US10144978B2 (en) | 2009-11-11 | 2012-05-10 | Probiotic bifidobacterium strain |
US15/936,661 Active US10597739B2 (en) | 2009-11-11 | 2018-03-27 | Probiotic bifidobacterium strain |
US16/789,768 Active US11225641B2 (en) | 2009-11-11 | 2020-02-13 | Probiotic Bifidobacterium strain |
US17/545,148 Abandoned US20220169976A1 (en) | 2009-11-11 | 2021-12-08 | Probiotic bifidobacterium strain |
Family Applications Before (3)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US13/468,374 Active 2031-02-19 US10144978B2 (en) | 2009-11-11 | 2012-05-10 | Probiotic bifidobacterium strain |
US15/936,661 Active US10597739B2 (en) | 2009-11-11 | 2018-03-27 | Probiotic bifidobacterium strain |
US16/789,768 Active US11225641B2 (en) | 2009-11-11 | 2020-02-13 | Probiotic Bifidobacterium strain |
Country Status (11)
Country | Link |
---|---|
US (4) | US10144978B2 (en) |
EP (1) | EP2498789B1 (en) |
CN (2) | CN102946891B (en) |
AU (2) | AU2010317422A1 (en) |
BR (1) | BR112012011315B1 (en) |
CA (1) | CA2779418C (en) |
ES (1) | ES2595444T3 (en) |
MX (1) | MX2012005524A (en) |
PL (1) | PL2498789T3 (en) |
RU (1) | RU2546251C2 (en) |
WO (1) | WO2011058535A1 (en) |
Families Citing this family (107)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
GB2460781B (en) * | 2007-03-28 | 2012-01-25 | Alimentary Health Ltd | Probiotic bifidobacterium strains |
US9457077B2 (en) | 2009-11-18 | 2016-10-04 | Katherine Rose Kovarik | Method and system for targeting the microbiome to promote health and treat allergic and inflammatory diseases |
US9585920B2 (en) | 2011-02-04 | 2017-03-07 | Katherine Rose Kovarik | Method and system for treating cancer cachexia |
US11357722B2 (en) | 2011-02-04 | 2022-06-14 | Seed Health, Inc. | Method and system for preventing sore throat in humans |
US10086018B2 (en) | 2011-02-04 | 2018-10-02 | Joseph E. Kovarik | Method and system for reducing the likelihood of colorectal cancer in a human being |
US10245288B2 (en) | 2011-02-04 | 2019-04-02 | Joseph E. Kovarik | Method and system for reducing the likelihood of developing NASH in an individual diagnosed with non-alcoholic fatty liver disease |
US11951139B2 (en) | 2015-11-30 | 2024-04-09 | Seed Health, Inc. | Method and system for reducing the likelihood of osteoporosis |
US9730967B2 (en) | 2011-02-04 | 2017-08-15 | Katherine Rose Kovarik | Method and system for treating cancer cachexia |
US10010568B2 (en) | 2011-02-04 | 2018-07-03 | Katherine Rose Kovarik | Method and system for reducing the likelihood of a spirochetes infection in a human being |
US11998479B2 (en) | 2011-02-04 | 2024-06-04 | Seed Health, Inc. | Method and system for addressing adverse effects on the oral microbiome and restoring gingival health caused by sodium lauryl sulphate exposure |
US10687975B2 (en) | 2011-02-04 | 2020-06-23 | Joseph E. Kovarik | Method and system to facilitate the growth of desired bacteria in a human's mouth |
US10940169B2 (en) | 2015-11-30 | 2021-03-09 | Joseph E. Kovarik | Method for reducing the likelihood of developing cancer in an individual human being |
US10512661B2 (en) | 2011-02-04 | 2019-12-24 | Joseph E. Kovarik | Method and system for reducing the likelihood of developing liver cancer in an individual diagnosed with non-alcoholic fatty liver disease |
US10314865B2 (en) | 2011-02-04 | 2019-06-11 | Katherine Rose Kovarik | Method and system for treating cancer and other age-related diseases by extending the healthspan of a human |
US10835560B2 (en) | 2013-12-20 | 2020-11-17 | Joseph E. Kovarik | Reducing the likelihood of skin cancer in an individual human being |
US9987224B2 (en) | 2011-02-04 | 2018-06-05 | Joseph E. Kovarik | Method and system for preventing migraine headaches, cluster headaches and dizziness |
US10111913B2 (en) | 2011-02-04 | 2018-10-30 | Joseph E. Kovarik | Method of reducing the likelihood of skin cancer in an individual human being |
US11419903B2 (en) | 2015-11-30 | 2022-08-23 | Seed Health, Inc. | Method and system for reducing the likelihood of osteoporosis |
US11951140B2 (en) | 2011-02-04 | 2024-04-09 | Seed Health, Inc. | Modulation of an individual's gut microbiome to address osteoporosis and bone disease |
US10548761B2 (en) | 2011-02-04 | 2020-02-04 | Joseph E. Kovarik | Method and system for reducing the likelihood of colorectal cancer in a human being |
US11844720B2 (en) | 2011-02-04 | 2023-12-19 | Seed Health, Inc. | Method and system to reduce the likelihood of dental caries and halitosis |
US11273187B2 (en) | 2015-11-30 | 2022-03-15 | Joseph E. Kovarik | Method and system for reducing the likelihood of developing depression in an individual |
US10085938B2 (en) | 2011-02-04 | 2018-10-02 | Joseph E. Kovarik | Method and system for preventing sore throat in humans |
US10583033B2 (en) | 2011-02-04 | 2020-03-10 | Katherine Rose Kovarik | Method and system for reducing the likelihood of a porphyromonas gingivalis infection in a human being |
US10842834B2 (en) | 2016-01-06 | 2020-11-24 | Joseph E. Kovarik | Method and system for reducing the likelihood of developing liver cancer in an individual diagnosed with non-alcoholic fatty liver disease |
US11523934B2 (en) | 2011-02-04 | 2022-12-13 | Seed Health, Inc. | Method and system to facilitate the growth of desired bacteria in a human's mouth |
US11191665B2 (en) | 2011-02-04 | 2021-12-07 | Joseph E. Kovarik | Method and system for reducing the likelihood of a porphyromonas gingivalis infection in a human being |
GB201112091D0 (en) | 2011-07-14 | 2011-08-31 | Gt Biolog Ltd | Bacterial strains isolated from pigs |
US10307167B2 (en) | 2012-12-14 | 2019-06-04 | Corquest Medical, Inc. | Assembly and method for left atrial appendage occlusion |
US10314594B2 (en) | 2012-12-14 | 2019-06-11 | Corquest Medical, Inc. | Assembly and method for left atrial appendage occlusion |
US10813630B2 (en) | 2011-08-09 | 2020-10-27 | Corquest Medical, Inc. | Closure system for atrial wall |
GB201117313D0 (en) | 2011-10-07 | 2011-11-16 | Gt Biolog Ltd | Bacterium for use in medicine |
FI126711B (en) | 2011-10-12 | 2017-04-13 | Gut Guide Oy | Assessing the health risk associated with a serotonin deficiency |
US20140142689A1 (en) | 2012-11-21 | 2014-05-22 | Didier De Canniere | Device and method of treating heart valve malfunction |
WO2014154493A1 (en) * | 2013-03-28 | 2014-10-02 | Nestec S.A. | Indoxyl sulfate as a biomarker of prebiotic efficacy for weight gain prevention |
GB201306536D0 (en) | 2013-04-10 | 2013-05-22 | Gt Biolog Ltd | Polypeptide and immune modulation |
US9566443B2 (en) | 2013-11-26 | 2017-02-14 | Corquest Medical, Inc. | System for treating heart valve malfunction including mitral regurgitation |
US11826388B2 (en) | 2013-12-20 | 2023-11-28 | Seed Health, Inc. | Topical application of Lactobacillus crispatus to ameliorate barrier damage and inflammation |
US11642382B2 (en) | 2013-12-20 | 2023-05-09 | Seed Health, Inc. | Method for treating an individual suffering from bladder cancer |
US11672835B2 (en) | 2013-12-20 | 2023-06-13 | Seed Health, Inc. | Method for treating individuals having cancer and who are receiving cancer immunotherapy |
US11529379B2 (en) | 2013-12-20 | 2022-12-20 | Seed Health, Inc. | Method and system for reducing the likelihood of developing colorectal cancer in an individual human being |
US11026982B2 (en) | 2015-11-30 | 2021-06-08 | Joseph E. Kovarik | Method for reducing the likelihood of developing bladder or colorectal cancer in an individual human being |
US11969445B2 (en) | 2013-12-20 | 2024-04-30 | Seed Health, Inc. | Probiotic composition and method for controlling excess weight, obesity, NAFLD and NASH |
US11980643B2 (en) | 2013-12-20 | 2024-05-14 | Seed Health, Inc. | Method and system to modify an individual's gut-brain axis to provide neurocognitive protection |
US11213552B2 (en) | 2015-11-30 | 2022-01-04 | Joseph E. Kovarik | Method for treating an individual suffering from a chronic infectious disease and cancer |
US12005085B2 (en) | 2013-12-20 | 2024-06-11 | Seed Health, Inc. | Probiotic method and composition for maintaining a healthy vaginal microbiome |
US11998574B2 (en) | 2013-12-20 | 2024-06-04 | Seed Health, Inc. | Method and system for modulating an individual's skin microbiome |
US11833177B2 (en) | 2013-12-20 | 2023-12-05 | Seed Health, Inc. | Probiotic to enhance an individual's skin microbiome |
US11839632B2 (en) | 2013-12-20 | 2023-12-12 | Seed Health, Inc. | Topical application of CRISPR-modified bacteria to treat acne vulgaris |
EP3097183A1 (en) | 2014-01-24 | 2016-11-30 | The Procter & Gamble Company | Web comprising a microorganism-containing fibrous element and method for making same |
US20150209468A1 (en) | 2014-01-24 | 2015-07-30 | The Procter & Gamble Company | Hygiene article containing microorganism |
EP3097182A1 (en) | 2014-01-24 | 2016-11-30 | The Procter & Gamble Company | Filaments comprising a microorganism and method for making same |
ES2768690T3 (en) * | 2014-08-29 | 2020-06-23 | Chr Hansen As | Probiotic strains of Bifidobacterium adolescentis |
US10842626B2 (en) | 2014-12-09 | 2020-11-24 | Didier De Canniere | Intracardiac device to correct mitral regurgitation |
KR20170091157A (en) | 2014-12-23 | 2017-08-08 | 4디 파마 리서치 리미티드 | Pirin polypeptide and immune modulation |
SI3065748T1 (en) | 2014-12-23 | 2018-05-31 | 4D Pharma Research Limited | A bacteroides thetaiotaomicron strain and its use in reducing inflammation |
PE20180673A1 (en) * | 2015-06-01 | 2018-04-19 | Univ Chicago | TREATMENT AGAINST CANCER BY MANIPULATING COMENSAL MICROFLORA |
US20160354507A1 (en) | 2015-06-07 | 2016-12-08 | The Procter & Gamble Company | Article of commerce containing absorbent article |
MA41010B1 (en) | 2015-06-15 | 2020-01-31 | 4D Pharma Res Ltd | Compositions comprising bacterial strains |
AU2016278067B2 (en) | 2015-06-15 | 2022-09-22 | Cj Bioscience, Inc. | Compositions comprising bacterial strains |
KR20180012849A (en) | 2015-06-15 | 2018-02-06 | 4디 파마 리서치 리미티드 | Composition Containing Bacterial Strain |
MA41060B1 (en) | 2015-06-15 | 2019-11-29 | 4D Pharma Res Ltd | Compositions comprising bacterial strains |
ES2742514T3 (en) | 2015-06-15 | 2020-02-14 | 4D Pharma Res Ltd | Compositions comprising bacterial strains |
US20170020750A1 (en) | 2015-07-23 | 2017-01-26 | The Procter & Gamble Company | Patch containing microorganism |
AU2016315266B2 (en) * | 2015-08-31 | 2022-12-22 | Société des Produits Nestlé S.A. | Methods and compositions using Bifidobacterium longum to treat or prevent depressive symptoms |
JP2018528938A (en) * | 2015-08-31 | 2018-10-04 | ネステク ソシエテ アノニム | Methods and compositions using Bifidobacterium longum for controlling emotional responses and for the treatment and prevention of potential mood modulation |
BR112018001179A2 (en) * | 2015-08-31 | 2018-09-11 | Nestec Sa | Methods and compositions using bifidobacterium longum to optimize breastfeeding |
KR20170032815A (en) * | 2015-09-15 | 2017-03-23 | 경희대학교 산학협력단 | Novel lactic acid bacteria and composition for preventing, improving or treating neurodegenerative diseases or cognitive dysfunction |
PT3209310T (en) | 2015-11-20 | 2018-04-20 | 4D Pharma Res Ltd | Compositions comprising bacterial strains |
GB201520497D0 (en) | 2015-11-20 | 2016-01-06 | 4D Pharma Res Ltd | Compositions comprising bacterial strains |
GB201520638D0 (en) | 2015-11-23 | 2016-01-06 | 4D Pharma Res Ltd | Compositions comprising bacterial strains |
GB201520631D0 (en) | 2015-11-23 | 2016-01-06 | 4D Pharma Res Ltd | Compositions comprising bacterial strains |
US10568916B2 (en) | 2015-11-30 | 2020-02-25 | Joseph E. Kovarik | Method and system for protecting honey bees, bats and butterflies from neonicotinoid pesticides |
US10086024B2 (en) | 2015-11-30 | 2018-10-02 | Joseph E. Kovarik | Method and system for protecting honey bees, bats and butterflies from neonicotinoid pesticides |
US11529412B2 (en) | 2015-11-30 | 2022-12-20 | Seed Health, Inc. | Method and system for protecting honey bees from pesticides |
US10675347B2 (en) | 2015-11-30 | 2020-06-09 | Joseph E. Kovarik | Method and system for protecting honey bees from fipronil pesticides |
US10933128B2 (en) | 2015-11-30 | 2021-03-02 | Joseph E. Kovarik | Method and system for protecting honey bees from pesticides |
BR112018011665A2 (en) * | 2015-12-11 | 2018-12-04 | Alimentary Health Ltd | bifidobacterium longum for the treatment of obesity and associated metabolic disorders |
GB201612191D0 (en) | 2016-07-13 | 2016-08-24 | 4D Pharma Plc | Compositions comprising bacterial strains |
CA3016179C (en) | 2016-03-04 | 2019-08-13 | 4D Pharma Plc | Compositions comprising bacterial blautia strains for treating visceral hypersensitivity |
TWI802545B (en) | 2016-07-13 | 2023-05-21 | 英商4D製藥有限公司 | Compositions comprising bacterial strains |
IT201600084470A1 (en) | 2016-08-10 | 2018-02-10 | Probiotical Spa | Composition for use in the treatment of major depressive disorder |
GB201621123D0 (en) | 2016-12-12 | 2017-01-25 | 4D Pharma Plc | Compositions comprising bacterial strains |
RU2761873C2 (en) | 2016-12-23 | 2021-12-13 | Кэйо Юниверсити | Compositions and methods for inducing cd8+ t-cells |
MX2019009079A (en) * | 2017-01-31 | 2019-09-26 | Univ Industry Cooperation Group Kyung Hee Univ | Novel lactic acid bacteria and use thereof. |
EP3589726A1 (en) * | 2017-02-28 | 2020-01-08 | Alimentary Health Limited | Bifidobacterium longum able to beneficially modulate immune response to respiratory virus infection |
EP3532603B1 (en) * | 2017-02-28 | 2020-09-09 | PrecisionBiotics Group Limited | Bifidobacterium longum able to beneficially modulate immune response to respiratory virus infection |
EP3630137B1 (en) | 2017-05-22 | 2023-05-17 | 4D Pharma Research Limited | Compositions comprising bacterial strains |
MA41708A (en) | 2017-05-24 | 2020-04-08 | 4D Pharma Res Ltd | COMPOSITIONS CONTAINING BACTERIAL STRAINS |
ES2855701T3 (en) | 2017-06-14 | 2021-09-24 | 4D Pharma Res Ltd | Compositions comprising bacterial strains |
MD3600364T2 (en) | 2017-06-14 | 2020-12-31 | 4D Pharma Res Ltd | Compositions comprising a bacterial strain of the genus Megasphaera and uses thereof |
ES2841902T3 (en) | 2017-06-14 | 2021-07-12 | 4D Pharma Res Ltd | Compositions comprising bacterial strains |
WO2019010255A1 (en) | 2017-07-05 | 2019-01-10 | Evelo Biosciences, Inc. | Compositions and methods of treating cancer using bifidobacterium animalis ssp. lactis |
CN109200064A (en) * | 2017-07-07 | 2019-01-15 | 中国农业大学 | The purposes of animal bifidobacteria A6 in medicine preparation |
ES2899674T3 (en) | 2018-01-29 | 2022-03-14 | Prec Group Limited | A combination product for the prophylaxis and treatment of irritable bowel syndrome |
KR20200116106A (en) * | 2018-01-29 | 2020-10-08 | 프리시젼바이오틱스 그룹 리미티드 | Combination products for the prevention and treatment of irritable bowel syndrome |
CN111918663B (en) * | 2018-01-29 | 2024-10-11 | 精密生物集团有限公司 | Bifidobacterium longum NCIMB 41676 |
WO2019145574A1 (en) | 2018-01-29 | 2019-08-01 | Alimentary Health Limited | Bifidobacterium longum ncimb 41676 |
EP3746097A1 (en) | 2018-01-29 | 2020-12-09 | PrecisionBiotics Group Limited | Bifidobacterium longum |
WO2019145573A1 (en) * | 2018-01-29 | 2019-08-01 | Alimentary Health Limited | Bifidobacterium longum ncimb 41676 |
JP2021512638A (en) | 2018-02-09 | 2021-05-20 | 学校法人慶應義塾 | Compositions and Methods for Inducing CD8 + T Cells |
WO2020063553A1 (en) | 2018-09-30 | 2020-04-02 | 内蒙古伊利实业集团股份有限公司 | Bifidobacterium lactis bl-99 and application thereof |
CN110964656B (en) * | 2018-09-30 | 2021-05-28 | 内蒙古伊利实业集团股份有限公司 | Bifidobacterium lactis BL-99 capable of preventing osteoporosis and application thereof |
CN109929773B (en) * | 2019-01-10 | 2020-06-19 | 江苏德禧生物科技有限公司 | Bifidobacterium capable of being used for selenium-rich culture and active protein and application thereof |
CN110974917A (en) * | 2019-11-25 | 2020-04-10 | 垒途智能教科技术研究院江苏有限公司 | Compound preparation with auxiliary effect of preventing and treating psychological depression of children and teenagers |
CN113209139B (en) * | 2020-08-31 | 2022-09-23 | 内蒙古蒙牛乳业(集团)股份有限公司 | Application of bifidobacterium lactis MN-Gup in improving obesity and characteristic intestinal flora thereof |
CN114381390B (en) * | 2021-12-20 | 2023-11-17 | 美益添生物医药(武汉)有限公司 | Bifidobacterium longum ME-875 and application thereof |
Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US10689719B2 (en) * | 2008-11-11 | 2020-06-23 | Precisionbiotics Group Limited | Bifidobacterium longum |
Family Cites Families (8)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPS55113718A (en) | 1979-02-27 | 1980-09-02 | Yakult Honsha Co Ltd | Antitumor agent |
ID29150A (en) * | 1999-01-15 | 2001-08-02 | Entpr Ireland Cs | USE OF LACTOBACILLUS SALIVARIUS |
US20030092163A1 (en) * | 2001-07-26 | 2003-05-15 | Collins John Kevin | Probiotic bifidobacterium strains |
US20040265279A1 (en) | 2003-05-08 | 2004-12-30 | Timothy Dinan | Probiotics in the treatment of atypical depression and other disorders characterized by hypothalamic pitiuitary-adrenal axis over-activity |
SE529185C2 (en) | 2005-10-07 | 2007-05-22 | Arla Foods Amba | Use of probiotic bacteria for the manufacture of food or drugs for the prevention of obesity |
GB2460781B (en) * | 2007-03-28 | 2012-01-25 | Alimentary Health Ltd | Probiotic bifidobacterium strains |
US8557233B2 (en) * | 2007-03-28 | 2013-10-15 | Alimentary Heath Limited | Probiotic bifidobacterium strains |
EP2110028A1 (en) * | 2008-04-15 | 2009-10-21 | Nestec S.A. | Bifidobacterium longum and hippocampal BDNF expression |
-
2010
- 2010-11-11 MX MX2012005524A patent/MX2012005524A/en not_active Application Discontinuation
- 2010-11-11 CN CN201080051187.8A patent/CN102946891B/en active Active
- 2010-11-11 RU RU2012117973/10A patent/RU2546251C2/en active
- 2010-11-11 EP EP10779340.8A patent/EP2498789B1/en active Active
- 2010-11-11 AU AU2010317422A patent/AU2010317422A1/en not_active Abandoned
- 2010-11-11 WO PCT/IE2010/000066 patent/WO2011058535A1/en active Application Filing
- 2010-11-11 ES ES10779340.8T patent/ES2595444T3/en active Active
- 2010-11-11 PL PL10779340T patent/PL2498789T3/en unknown
- 2010-11-11 BR BR112012011315-1A patent/BR112012011315B1/en active IP Right Grant
- 2010-11-11 CN CN201710691551.0A patent/CN107574131A/en active Pending
- 2010-11-11 CA CA2779418A patent/CA2779418C/en active Active
-
2012
- 2012-05-10 US US13/468,374 patent/US10144978B2/en active Active
-
2015
- 2015-02-19 AU AU2015200833A patent/AU2015200833B2/en active Active
-
2018
- 2018-03-27 US US15/936,661 patent/US10597739B2/en active Active
-
2020
- 2020-02-13 US US16/789,768 patent/US11225641B2/en active Active
-
2021
- 2021-12-08 US US17/545,148 patent/US20220169976A1/en not_active Abandoned
Patent Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US10689719B2 (en) * | 2008-11-11 | 2020-06-23 | Precisionbiotics Group Limited | Bifidobacterium longum |
Also Published As
Publication number | Publication date |
---|---|
WO2011058535A1 (en) | 2011-05-19 |
CN102946891A (en) | 2013-02-27 |
US11225641B2 (en) | 2022-01-18 |
PL2498789T3 (en) | 2017-01-31 |
US10144978B2 (en) | 2018-12-04 |
RU2012117973A (en) | 2013-12-20 |
AU2015200833A1 (en) | 2015-03-12 |
CA2779418A1 (en) | 2011-05-19 |
AU2010317422A1 (en) | 2012-05-31 |
MX2012005524A (en) | 2013-04-03 |
EP2498789B1 (en) | 2016-06-22 |
CA2779418C (en) | 2018-03-20 |
US20180202010A9 (en) | 2018-07-19 |
EP2498789A1 (en) | 2012-09-19 |
RU2546251C2 (en) | 2015-04-10 |
ES2595444T3 (en) | 2016-12-30 |
US20120276143A1 (en) | 2012-11-01 |
BR112012011315A2 (en) | 2020-08-25 |
BR112012011315B1 (en) | 2022-03-22 |
US20200325548A1 (en) | 2020-10-15 |
US20180282825A1 (en) | 2018-10-04 |
CN107574131A (en) | 2018-01-12 |
US10597739B2 (en) | 2020-03-24 |
CN102946891B (en) | 2017-09-01 |
AU2015200833B2 (en) | 2017-03-16 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
US20220169976A1 (en) | Probiotic bifidobacterium strain | |
US8709398B2 (en) | Probiotic Bifidobacterium strains | |
US8557233B2 (en) | Probiotic bifidobacterium strains | |
EP1409645B1 (en) | Probiotic lactobacillus salivarius strains | |
US20060121015A1 (en) | Probiotic bifidobacterium strains | |
US20120207713A1 (en) | Probiotic bifidobacterium strains | |
IE20080245A1 (en) | Probiotic Bifidobacteria strains | |
IE20080244A1 (en) | Probiotic Bifidobacteria strains | |
IE86044B1 (en) | Probiotic bifidobacteria strains | |
IE86045B1 (en) | Probiotic bifidobacteria strains | |
ZA200400555B (en) | Probiotic bifidobacterium strains. |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
STPP | Information on status: patent application and granting procedure in general |
Free format text: DOCKETED NEW CASE - READY FOR EXAMINATION |
|
AS | Assignment |
Owner name: PRECISIONBIOTICS GROUP LIMITED, IRELAND Free format text: CHANGE OF NAME;ASSIGNOR:ALIMENTARY HEALTH LIMITED;REEL/FRAME:062749/0689 Effective date: 20191216 Owner name: UNIVERSITY COLLEGE CORK - NATIONAL UNIVERSITY OF IRELAND, CORK, IRELAND Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNOR:DINAN, TIMOTHY G.;REEL/FRAME:062749/0686 Effective date: 20101101 Owner name: UNIVERSITY COLLEGE CORK - NATIONAL UNIVERSITY OF IRELAND, CORK, IRELAND Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNOR:CRYAN, JOHN F.;REEL/FRAME:062749/0683 Effective date: 20101101 Owner name: ALIMENTARY HEALTH LIMITED, IRELAND Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNOR:THE PROCTER & GAMBLE COMPANY;REEL/FRAME:062696/0507 Effective date: 20170306 Owner name: THE PROCTER & GAMBLE COMPANY, OHIO Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNOR:ALIMENTARY HEALTH LIMITED;REEL/FRAME:062696/0504 Effective date: 20120629 Owner name: ALIMENTARY HEALTH LIMITED, IRELAND Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNOR:UNIVERSITY COLLEGE CORK - NATIONAL UNIVERSITY OF IRELAND, CORK;REEL/FRAME:062696/0496 Effective date: 20101110 Owner name: ALIMENTARY HEALTH LIMITED, IRELAND Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNORS:O'MAHONY, LIAM;KIELY, BARRY;MURPHY, EILEEN FRANCES;REEL/FRAME:062696/0486 Effective date: 20101110 |
|
STPP | Information on status: patent application and granting procedure in general |
Free format text: NON FINAL ACTION MAILED |
|
STCB | Information on status: application discontinuation |
Free format text: ABANDONED -- FAILURE TO RESPOND TO AN OFFICE ACTION |