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CN1186696A - Application of beneficial bacteria in improving specific immunity of organism specific and in development of vaccine - Google Patents

Application of beneficial bacteria in improving specific immunity of organism specific and in development of vaccine Download PDF

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CN1186696A
CN1186696A CN97125843A CN97125843A CN1186696A CN 1186696 A CN1186696 A CN 1186696A CN 97125843 A CN97125843 A CN 97125843A CN 97125843 A CN97125843 A CN 97125843A CN 1186696 A CN1186696 A CN 1186696A
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immunity
group
bacterium
bacillus
bifurcated
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徐建国
林纪胜
蓝景刚
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Institute of Microbiology and Epidemiology of AMMS
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Institute of Microbiology and Epidemiology of AMMS
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Abstract

To the organism obtaining initial imunization through hypodermic vaccine injection, oral taking or natural infection, beneficial bacteria, such as bifidobacterium, lactobacillus acidophilus and colibacillus expressing protective antigen gene, are used separately or compositely for immune regulation in the same way as the first time so as to raise specific immunity level and protecting capacity. The result is compared with check group and shows obvious diffence (P<0.05). The said immune method is safe and reliable and has great application value in preventing and controlling infectious disease.

Description

Probiotic bacteria is improving specific immunity of organism specific and the developing application of epidemic disease (bacterium) Seedling
The present invention relates to the escherichia coli of probiotic bacteria such as bacillus bifidus, bacillus acidophilus and expression protective antigen gene; be particularly related to and use them to improve the specific immunity level that obtains the initial immunity body through epidemic disease (bacterium) Seedling initial immunity or natural infection, and their application aspect development of epidemic disease (bacterium) Seedling and immunization strategy.
Ideal epidemic disease (bacterium) Seedling should meet two conditions: the one, and safety is good, and the 2nd, have good immunogenicity.Both normally a pair of contradictions, otherwise safety is better, and immunogenicity is relatively poor; Immunogenicity is better, and toxic and side effects is stronger.This contradiction has limited application and the development in practice of epidemic disease (bacterium) Seedling.At present, the immunization scheme of epidemic disease (bacterium) Seedling mostly is first primary vaccination, carries out booster immunization again through behind the certain hour.Existing to this immunization strategy degree varies complicated operation, cost increases, uses and bother especially that the child is difficult to shortcomings such as acceptance.
Bacillus bifidus and bacillus acidophilus are the important members of normal flora in the humans and animals intestinal, at infection, regulate immunity of organism, defying age, antitumor and provide aspect such as some nutrient substance to play important effect.The normal escherichia coli of intestinal also are one of normal flora members, and it is widely used in the research of epidemic disease (bacterium) Seedling as the carrier of protective antigen gene.But the escherichia coli that utilize probiotic bacteria such as bacillus bifidus, bacillus acidophilus and expression protective antigen gene involved in the present invention are improved the immune programme for children and the strategy of body specificity level and immunity, and report is not at home and abroad arranged.
The present invention's purpose; be the escherichia coli that utilize probiotic bacteria such as bifurcated bacillus bifidus, bacillus acidophilus and expression epidemic disease (bacterium) Seedling protective antigen gene; to body through epidemic disease (bacterium) Seedling subcutaneous injection and oral route initial immunity; or natural infection obtains the body of initial immunity; through behind the certain hour; by the approach identical with initial immunity; respectively with its different combination the initial immunity body is carried out immunomodulating; solve after epidemic disease (bacterium) Seedling initial immunity or natural infection acquisition initial immunity the problem that specific immunity descends and the protection period is short.
Subject matter of the present invention and technical scheme are as follows:
1. select SFF (Specific pathogen free SPF) animal BALB/c mouse (available from Ministry of Public Health Beijing pharmaceutical biological product calibrating institute) for use, male and female half and half, random packet, male and female divide box to raise.
2. strain derives from Inst. of Epidemiology and Microbiology, Chinese Academy of Preventive Medicine (, strain principal character (seeing attached list 1).
The principal character of subordinate list 1. strains
The title principal character
Fu Shi 2a shigella 301 wild type strains
Dysentery I type shigella 112 wild type strains
Two valency (Fu Shi 2a, the dysentery I type shigella) vaccines of DOM3*;
ThyA-; The antibiotic-free resistance
E.coli X511 expresses the escherichia coli of dysentery I type shigella lipopolysaccharide
The bifurcated bacillus bifidus separates from normal children feces
The bacillus acidophilus
E.coli LE392 engineering strain
* two valency vaccine candidate strain DOM3 of shigella: with the strain of Fu Shi 2a shigella thy A genetic flaw is recipient bacterium, changes gene constructed the forming of somatic antigen of dysentery I type shigella over to.
3. antibacterial culturing: two valency vaccine candidate strain DOM3 of shigella and E.coli.X511, take out recovery from-70 ℃ of refrigerators respectively and cultivate, the single bacterium colony of picking, (NaCl 10g, yeast powder 5g, peptone 10g, adding distil water be to 1000ml, PH7.4) culture medium with LB; Wild type strain Fu Shi 2a shigella 301 and dysentery I type shigella 112 are taken out recovery from-70 ℃ of refrigerators respectively and cultivate, and the single bacterium colony of picking is with PA (Carnis Bovis seu Bubali cream 1.5g, NaCl 3.5g, K 2HPO 4.3H 2O 4.8g, KH 2PO 41.32g tryptone 5g, glucose 1g, yeast powder 1.5g, adding distil water be to 1000ml, PH7.2) culture medium; 37 ℃, 160 rev/mins incubated overnight are done 50 times of dilutions next day in shaking table, continue to cultivate 2-3 hour.The bifurcated bacillus bifidus is cultivated from freeze-drying lactobacillus pipe recovery back anaerobism, and the single bacterium colony of picking is with BL (Carnis Bovis seu Bubali cream 3g, polypepton 10g, tryptone 3g plants peptone 3g, glucose 10g, yeast powder 5g, soluble starch 0.5g, Tween 80 1ml, 5% cysteine 10ml, liver soup 150ml, Sanguis caprae seu ovis 50ml, agar 15g, solution A 10ml, solution B .5ml, adding distil water be to 1000ml, PH7.2; Solution A: KH 2PO 425g, K 2HPO 425g, adding distil water is to 250ml; Solution B: MgSO 4.7H 2O 10g, FeSO 4.7H 2O 0.5g, NaCl 0.5g, MnSO 40.337g adding distil water is to 250ml) culture medium, in anaerobic jar, cultivated 48-72 hour for 37 ℃.The bacillus acidophilus takes out recovery from-70 ℃ of refrigerators and cultivates, and the single bacterium colony of picking is with MRS (peptone 10g, Carnis Bovis seu Bubali cream 10g, yeast powder 5g, glucose 20g, K 2HPO 42.0g, sodium acetate 5.0g, Chinese holly edge acid three ammonia 2.0g, Tween 80 1ml, MgSO 4.7H 2O 200mg, MnSO 4.4H 2O 50mg, adding distil water are to 1000ml, and PH6.0) culture medium standing over night in 37 ℃ of incubators is cultivated.Collect thalline, with normal saline (subcutaneous injection immunity) or 0.2mol/LNaHCO 3(oral immunity) is mixed with needed bacteria concentration.
4. percutaneous is injected initial immunity and immunoregulatory program down: vaccine candidate strain DOM3 subcutaneous injection initial immunity program (seeing attached list 2); Subcutaneous injection immunomodulating program (seeing attached list 3), the subcutaneous injection initial immunity is after 6 weeks (i.e. in the 7th week) the last time, by the subcutaneous injection approach, give bifurcated bacillus bifidus, combination that the bacillus acidophilus is different with it with E.coli X511 respectively, the mice through initial immunity is carried out immunomodulating.
Subordinate list 2. subcutaneous injection initial immunity programs
Position number of times next day of between bacterium name bacterium amount (CFU)/0.1ml
DOM3 10 83 groin subcutaneous 3
Subordinate list 3. subcutaneous injection immunomodulating programs
Position number of times next day of between group bacterium amount CFU/0.1ml
Bifurcated bacillus bifidus 10 103 groin subcutaneous 3
The bacillus acidophilus 10 103 groin subcutaneous 3
E.coli X511 10 93 groin subcutaneous 3
Bifurcated bacillus bifidus+bacillus acidophilus
Each is 10 years old 103 groin subcutaneous 3
Bifurcated bacillus bifidus+bacillus acidophilus+E.coli.X511
Each is 10 years old 10+ 10 93 groin subcutaneous 3
Normal saline 0.1ml 3 groin subcutaneous 3
5. oral administration initial immunity and immunomodulating and antibody detection program (seeing attached list 4).Wherein: S, blood 50 μ l are got in docking, dilute 20 times, the centrifuging and taking dilute serum; F gets stool in mice, dilutes 10 times, puts 4 ℃ of soaked overnight, through 12, and 000rpm * 10 minute centrifuging and taking supernatant; V, per os pour into the two valency vaccine candidate strain DOM3 10 of shigella 9/ ml * 0.5ml; N, per os pour into 0.5ml 0.2mol/L NaHCO 3Solution; X, per os pour into E.coli X511 bacterium 10 9/ ml * 0.5ml; L, per os pour into bacillus acidophilus 10 9/ ml * 0.5ml; B, per os pour into bifurcated bacillus bifidus 10 9/ ml * 0 5ml; M1, per os pour into bifurcated bacillus bifidus 10 9/ ml, the bacillus acidophilus 10 9/ ml equivalent mixed liquor 0.5ml; M2, per os pour into bifurcated bacillus bifidus 10 9/ ml, the bacillus acidophilus 10 9/ ml, E.coli X511 bacterium 10 9/ ml equivalent mixed liquor 0.5ml.All bacterium liquid are all used 0.2mol/L NaHCO 3Preparation.
Subordinate list 4. oral administration initial immunities and immunomodulating and antibody detection program Zu Bie days 037 14 30 33 37 44 60 I S, F, V S, F S, F S, F S, F, N S, F S, F S, F S, FII S, F, V S, F S, F S, F S, F, X S, F S, F S, F S, FIII S, F, V S, F S, F S, F S, F, L S, F S, F S, F S, FIV S, F, V S, F S, F S, F S, F, B S, F S, F S, F S, FV S, F, V S, F S, F S, F S, F, M1 S, F S, F S, F S, FVI S, F, V S, F S, F S, F S, F, M2 S, F S, F S, F S, FVII S, F, N S, F S, F S, F S, F, N S, F S, F S, F S, FVIII S, F S, F S, F S, F S, F S, F S, F S, F S, F
The method that detects of antibody titer (enzyme-linked immunosorbent assay Enzyme linked immunosorbent assay, ELISA): envelope antigen: Fu Shi 2a shigella 301 and dysentery I type shigella 112 thalline; Sheep anti mouse IgA, IgM (HRP) be available from Sigma company, and sheep anti-mouse igg (HRP) is available from magnificent biological product company.
7. antibody titer detection of dynamic: the last time behind the subcutaneous injection initial immunity the 1st week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks (in the 7th week, immunomodulating is carried out in subcutaneous injection bifurcated bacillus bifidus, bacillus acidophilus, E.coli X511 and different combinations thereof), 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, mice is taken a blood sample through eye socket, adopt 5-11 mouse orbit blood for every group, every Mus is all surveyed the titre of specific antibody IgA, IgM, IgG in its serum, the geometric mean of mice serum antibody titer is as the titre of corresponding antibodies in this group mice serum.
8. to the attack of mice after the subcutaneous injection immunomodulating: through the DOM3 initial immunity and before dropping near immunity without the serum antibody titer of immunomodulating mice during level (in the 9th week); attack mice after immunomodulating through the abdominal cavity with the wild type strain; observe the existence of every group of mice, dead situation, determine respectively to organize the difference of protection.
The laboratory observation result:
1. bifurcated bacillus bifidus, bacillus acidophilus, E.coli X511 are to the immunological enhancement (seeing accompanying drawing 1) through the SPF of initial immunity BALB/c mouse.
1). the immunogenicity of vaccine candidate strain DOM3: behind DOM3 subcutaneous injection initial immunity, antibody I gA and IgG titre peak value appear at the 2nd week behind the last subcutaneous injection in the mice serum, and IgM titre peak value appeared at for the 1st week: serum titer progressively descends then, subcutaneous injection tended towards stability after 6 weeks the last time approximately.The specific antibody that has anti-Fu Shi 2a shigella and anti-dysentery I type shigella in the mice serum simultaneously, and the trend basically identical of its titre variation.Illustrate that vaccine candidate strain DOM3 has good two valencys, but the antibody titer of anti-Fu Shi 2a shigella is higher than the antibody titer of anti-dysentery I type shigella, this may relevant with the characteristics of vaccine own (seeing accompanying drawing 2 (a) and (b), (c)).
2). percutaneous is injected the dynamic change (seeing accompanying drawing 3 (a) and (b), (c)) of specific antibody titre in the mice serum of immunomodulating front and back down.Mice is first week after last subcutaneous injection immunomodulating, IgG antibody for the anti-Fu Shi 2a of specificity in serum shigella, bifurcated bacillus bifidus group, bacillus acidophilus's group, E.coli X511 group, bifurcated bacillus bifidus+bacillus acidophilus's group and bifurcated bacillus bifidus+bacillus acidophilus+E.coli X511 group, immunological enhancement to mice behind the initial immunity, through one factor analysis of variance, and matched group is to compare between the normal saline group, all exist significant difference (P=0.0000,<0.05).IgG antibody for the anti-dysentery I type of specificity in serum shigella, identical with the IgG antibody of the anti-Fu Shi 2a of specificity in serum shigella, bifurcated bacillus bifidus group, bacillus acidophilus's group, E.coli X511 group, bifurcated bacillus bifidus+bacillus acidophilus's group and bifurcated bacillus bifidus+bacillus acidophilus+E.coli X511 group, immunological enhancement to mice behind the initial immunity, compare with matched group, also all exist significant difference (P=0.0000,<0.05).Result and the IgG antibody of specific antibody IgM, IgA are similar.It should be noted that E.coli X511 group and bifurcated bacillus bifidus+bacillus acidophilus+E.coliX51 group, may be because have the specific immunity of stimulation memory impairments, its specific antibody titre is high than other group.
3). percutaneous can improve the protective effect that the initial immunity mice is attacked shigella wild type strain after the injection immunomodulating down.Give bifurcated bacillus bifidus, bacillus acidophilus, E.coli X511 and bifurcated bacillus bifidus+bacillus acidophilus respectively, bifurcated bacillus bifidus+bacillus acidophilus+E.coli X511 carries out immunomodulating, mice after the immunomodulating is attacked in abdominal cavities respectively with wild strain Fu Shi 2a shigella 301 and dysentery I type shigella 112 in after the subcutaneous injection immunomodulating the 14th day the last time.Bifurcated bacillus bifidus group, bacillus acidophilus group, E.coli X511 group and bifurcated bacillus bifidus+bacillus acidophilus's group and bifurcated bacillus bifidus+bacillus acidophilus+E.coli X511 group, to mice with 234 LD 50(LD 50=2.2388 * 10 5CFU) wild type strain Fu Shi 2a shigella 301 and 184 LD 50(LD 50=2.9535 * 10 5CFU) protection that wild type strain dysentery I type shigella 112 is attacked is respectively compared with matched group, all has significant difference (P<0.05).The result shows, although antibody titer has height that the end is arranged, protects effect to reach unanimity.Bifurcated bacillus bifidus and bacillus acidophilus can be with its nonspecific immunity protective effects; be used for the raising of specificity protection; adopt this immunization strategy, nonspecific immunity protection and specific immunity protection can realize organically combining, and reach best protection effect (seeing attached list 5).
2. oral bifurcated bacillus bifidus, bacillus acidophilus and E.coli X511 and different combinations thereof can improve the specific antibody IgA titre of oral administration initial immunity mouse intestinal.The dynamic change (seeing accompanying drawing 4-9) of specific antibody in stool in mice, the serum before and after oral bifurcated bacillus bifidus, bacillus acidophilus and E.coli X511 and the different combination thereof.Oral bifurcated bacillus bifidus, bacillus acidophilus and E.coli X511 and different combinations thereof do not have remarkable potentiation to IgG antibody, the IgM of anti-Fu Shi 2a shigella in the oral administration initial immunity stool in mice and anti-dysentery I type shigella; Oral bifurcated bacillus bifidus, bacillus acidophilus and E.coli X511 and different combinations thereof, improved to degree varies the antibody I gA level of the specificity anti-Fu Shi 2a shigella and the anti-dysentery I type shigella of oral administration initial immunity stool in mice, bifurcated bacillus bifidus+bacillus acidophilus's group and bifurcated bacillus bifidus+bacillus acidophilus+E.coli X511 group is suitable with the peak value of E.coli X511 group; Bifurcated bacillus bifidus, bacillus acidophilus and E.coli X511 and different combinations thereof have certain potentiation to IgG antibody, the IgM of anti-Fu Shi 2a shigella in the oral administration initial immunity mice serum and anti-dysentery I type shigella.
Good effect of the present invention is; adopt this immunization strategy; can not only improve the specific immunity level and the protection of body; and it is safe and reliable; to utilize epidemic disease (bacterium) Seedling or in natural infection colony the prevention and control infectious disease aspect; have very big use value and commercial value, have a extensive future.
The mice of subordinate list 5. subcutaneous injection initial immunities is attacked bacterial strain attack bacterium amount death toll existence number P value * to the protection effect initial immunity bacterial strain immunomodulating bacterial strain of mice after with the approach immunomodulating
Fu Shi 2a shigella 301 234LD 5012 8<0.05 DOM3 bifurcated bacillus bifidus
Dysentery I type shigella 112 184LD 5016 4<0.05
Fu Shi 2a shigella 301 234LD 5015 5<0.05 DOM3 bacillus acidophilus
Dysentery I type shigella 112 184LD 5015 5<0.05
Fu Shi 2a shigella 301 234LD 5012 8<0.05 DOM3 E.coli X511
Dysentery I type shigella 112 184LD 508 12<0.05
Fu Shi 2a shigella 301 234LD 5012 8<0.05 DOM3 bifurcated bacillus bifidus+bacillus acidophilus
Dysentery I type shigella 112 184LD 5014 6<0.05
Fu Shi 2a shigella 301 234LD 5012 8<0.05 DOM3 bifurcated bacillus bifidus+bacillus acidophilus+E.coli X511
Dysentery I type shigella 112 184LD 508 12<0.05
Fu Shi 2a shigella 30 234LD 5020 0 DOM3 normal saline
Dysentery I type shigella 112 184LD 5020 0 * and normal saline group X 2Check.
Description of drawings:
At the beginning of the escherichia coli of accompanying drawing 1-probiotic bacteria such as bifurcated bacillus bifidus, bacillus acidophilus and expression protective antigen gene are improved
Inferior immune body specific antibody titre sketch map
Wherein:
Figure A9712584300071
-initial immunity
Figure A9712584300072
-probiotic bacteria is regulated and is injected specific antibody titre dynamic change figure in the initial immunity mice serum under the immune accompanying drawing 2-percutaneous
Wherein:
(a)-dynamic change of IgG titre:
(b)-dynamic change of IgM titre:
(c)-dynamic change of IgA titre.
Figure A9712584300081
The anti-Fu Shi 2a of anti-Sh.f2a-shigella antibody; The anti-dysentery of anti-Sh.dl-is hate I type shigella antibody.Accompanying drawing 3-percutaneous injection is down regulated specific antibody titre dynamic change figure in the mice serum of immunity back
Wherein, (a)-dynamic change of IgG titre;
(b)-dynamic change of IgM titre;
(c)-dynamic change of IgA titre. Anti-Fu Shi 2a shigella antibody in the A:anti-Sh.f2a-normal saline group;
Figure A9712584300084
Anti-dysentery I type shigella antibody in the A:anti-Sh.dl-normal saline group;
Figure A9712584300085
Anti-Fu Shi 2a shigella antibody in the B:anti-Sh.f2a-bifurcated bacillus bifidus group:
Figure A9712584300086
Anti-dysentery I type shigella antibody in the B:anti-Sh.dl-bifurcated bacillus bifidus group;
Figure A9712584300087
Anti-Fu Shi 2a shigella antibody in the C:anti-Sh.f2a-bacillus acidophilus group;
Figure A9712584300088
Anti-dysentery I type shigella antibody in the C:anti-Sh.dl-bacillus acidophilus group;
Figure A9712584300089
Anti-Fu Shi 2a shigella antibody in the D:anu-Sn.f2a-E.coli X511 group;
Figure A97125843000810
Anti-dysentery I type shigella antibody in the D:anti-Sh.dl-E.coli X511 group;
Figure A97125843000811
Anti-Fu Shi 2a shigella antibody in E:anti-Sh.f2a-bifurcated bacillus bifidus+bacillus acidophilus's group;
Figure A97125843000812
Anti-dysentery I type shigella antibody in E:anti-Sh.dl-bifurcated bacillus bifidus+bacillus acidophilus's group; Anti-Fu Shi 2a shigella antibody in F:anti-Sh.f2a-bifurcated bacillus bifidus+bacillus acidophilus+E.coli X511 group;
Figure A97125843000814
Anti-dysentery I type shigella antibody in F:anti-Sh.dl-bifurcated bacillus bifidus+bacillus acidophilus+E.coli X511 group.Specific antibody titre dynamic change figure in the stool in mice before and after the immunomodulating of accompanying drawing 4-oral administration
Wherein: (a)-anti-dysentery hate the variation of I type shigella antibody I gA;
(b)-variation of anti-Fu Shi 2a shigella antibody I gA.
Figure A97125843000815
I: DOM3 group; II: DOM3+E.coli X511 group;
Figure A97125843000817
III: DOM3+ bacillus acidophilus group; IV: DOM3+ bifurcated bacillus bifidus group;
Figure A97125843000819
V: DOM3+ bacillus acidophilus+bifurcated bacillus bifidus group;
Figure A97125843000820
VI: DOM3+ bacillus acidophilus group+bifurcated bacillus bifidus+E.coli X511 group;
Figure A97125843000821
VI: NAHCO3 matched group; VIII: blank group.Specific antibody titre dynamic change figure in the stool in mice before and after the immunomodulating of accompanying drawing 5-oral administration
Wherein: (a)-anti-dysentery hate the variation of I type shigella IgG antibody;
(b)-variation of anti-Fu Shi 2a shigella IgG antibody.
Figure A97125843000823
I: DOM3 group;
Figure A97125843000824
II: DOM3+E.coli X511 group; III: DOM3+ bacillus acidophilus group; IV: DOM3+ bifurcated bacillus bifidus group;
Figure A97125843000827
V: DOM3+ bacillus acidophilus+bifurcated bacillus bifidus group;
Figure A97125843000828
VI: DOM3+ bacillus acidophilus group+bifurcated bacillus bifidus+E.coli X511 group;
Figure A97125843000829
VII: NAHCO3 matched group; VIII: blank group.Specific antibody titre dynamic change figure in the stool in mice before and after the immunomodulating of accompanying drawing 6-oral administration
Wherein: (a)-anti-dysentery hate the variation of I type shigella antibody IgM;
(b)-variation of anti-Fu Shi 2a shigella antibody IgM.
Figure A9712584300091
I: DOM3 group;
Figure A9712584300092
II: DOM3+E.coli X511 group;
Figure A9712584300093
III: DOM3+ bacillus acidophilus group;
Figure A9712584300094
IV: DOM3+ bifurcated bacillus bifidus group;
Figure A9712584300095
V: DOM3+ bacillus acidophilus+bifurcated bacillus bifidus group;
Figure A9712584300096
VI: DOM3+ bacillus acidophilus group+bifurcated bacillus bifidus+E.coli X511 group;
Figure A9712584300097
VII: NAHCO3 matched group;
Figure A9712584300098
VIII: blank group.Specific antibody titre dynamic change figure in the mice serum before and after the immunomodulating of accompanying drawing 7-oral administration
Wherein: (a)-anti-dysentery hate the variation of I type shigella IgG antibody;
(b)-variation of anti-Fu Shi 2a shigella IgG antibody.
Figure A9712584300099
I: DOM3 group; II: DOM3+E.coli X511 group; III: DOM3+ bacillus acidophilus group;
Figure A97125843000912
IV: DOM3+ bifurcated bacillus bifidus group;
Figure A97125843000913
V: DOM3+ bacillus acidophilus+bifurcated bacillus bifidus group;
Figure A97125843000914
VI: DOM3+ bacillus acidophilus group+bifurcated bacillus bifidus+E.coli X511 group;
Figure A97125843000915
VII: NAHCO3 matched group; VIII: blank group.Specific antibody titre dynamic change figure in the mice serum before and after the immunomodulating of accompanying drawing 8-oral administration
Wherein: (a)-anti-dysentery hate the variation of I type shigella antibody IgM;
(b)-variation of anti-Fu Shi 2a shigella antibody IgM.
Figure A97125843000917
I: DOM3 group;
Figure A97125843000918
II: DOM3+E.coli X511 group; III: DOM3+ bacillus acidophilus group; IV: DOM3+ bifurcated bacillus bifidus group; V: DOM3+ bacillus acidophilus+bifurcated bacillus bifidus group; VI: DOM3+ bacillus acidophilus group+bifurcated bacillus bifidus+E.coli X511 group; VII: NAHCO3 matched group; VIII: blank group.Specific antibody titre dynamic change figure in the mice serum before and after the immunomodulating of accompanying drawing 9-oral administration
Wherein: (a)-variation of anti-dysentery I type shigella antibody I gA;
(b)-variation of anti-Fu Shi 2a shigella antibody I gA.
Figure A97125843000925
I: DOM3 group;
Figure A97125843000926
II: DOM3+E.coli X511 group;
Figure A97125843000927
III: DOM3+ bacillus acidophilus group: IV: DOM3+ bifurcated bacillus bifidus group;
Figure A97125843000929
V: DOM3+ bacillus acidophilus+bifurcated bacillus bifidus group;
Figure A97125843000930
VI: DOM3+ bacillus acidophilus group+bifurcated bacillus bifidus+E.coli X511 group; VII: NAHCO3 matched group;
Figure A97125843000932
VIII: blank group.

Claims (5)

1. probiotic bacteria is improving specific immunity of organism specific and the developing application of epidemic disease (bacterium) Seedling; it is characterized in that body is carried out initial immunity by subcutaneous injection and oral route; or natural infection obtains initial immunity; through behind the certain hour; by the approach identical with initial immunity; give probiotic bacteria such as bifurcated bacillus bifidus, bacillus acidophilus and the escherichia coli of expression epidemic disease (bacterium) Seedling protective antigen gene and different combinations thereof respectively; body to initial immunity carries out immunomodulating, to improve the specific immunity level and the protection of body.
According to the described probiotic bacteria of claim 1 improving specific immunity of organism specific and the developing application of epidemic disease (bacterium) Seedling, it is characterized in that preparing epidemic disease (bacterium) Seedling, injection and oral route carry out initial immunity, or natural infection acquisition initial immunity down to the body percutaneous.
3 are improving specific immunity of organism specific and the developing application of epidemic disease (bacterium) Seedling according to claim 1 and 2 described probiotic bacterias; it is characterized in that behind initial immunity or natural infection acquisition initial immunity certain hour; the large intestine bar of preparation probiotic bacteria such as bifurcated bacillus bifidus, bacillus acidophilus and expression epidemic disease (bacterium) Seedling protective antigen gene; by the approach identical with initial immunity; give probiotic bacteria such as bifurcated bacillus bifidus, bacillus acidophilus and the large intestine bar of expression epidemic disease (bacterium) Seedling protective antigen gene and different combinations thereof respectively, the body of initial immunity is carried out immunomodulating.
4. according to claim 1,2 and 3 described probiotic bacterias are improving specific immunity of organism specific and the developing application of epidemic disease (bacterium) Seedling, it is characterized in that adopting enzyme-linked immunosorbent assay (ELISA) to detect serum and the specific antibody IgG in the feces, IgM and the IgA level of body after the probiotic bacteria immunomodulating.
According to the described probiotic bacteria of claim 1,2 and 3 improving specific immunity of organism specific and the developing application of epidemic disease (bacterium) Seedling, it is characterized in that the body after the probiotic bacteria immunomodulating, attack with the wild type strain, observe its protection effect.
CN97125843A 1997-12-25 1997-12-25 Application of beneficial bacteria in improving specific immunity of organism specific and in development of vaccine Pending CN1186696A (en)

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN100421676C (en) * 2003-12-17 2008-10-01 纽迪西亚公司 Lactic acid producing bacteria and lung function

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN100421676C (en) * 2003-12-17 2008-10-01 纽迪西亚公司 Lactic acid producing bacteria and lung function

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