FR2625100A2 - Use of antigens for the preparation of vaccines, protein rendered immunogenic and pharmaceutical composition containing it - Google Patents
Use of antigens for the preparation of vaccines, protein rendered immunogenic and pharmaceutical composition containing it Download PDFInfo
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- FR2625100A2 FR2625100A2 FR8718181A FR8718181A FR2625100A2 FR 2625100 A2 FR2625100 A2 FR 2625100A2 FR 8718181 A FR8718181 A FR 8718181A FR 8718181 A FR8718181 A FR 8718181A FR 2625100 A2 FR2625100 A2 FR 2625100A2
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/005—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from viruses
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/12—Viral antigens
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/12—Viral antigens
- A61K39/21—Retroviridae, e.g. equine infectious anemia virus
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
- G01N33/56983—Viruses
- G01N33/56988—HIV or HTLV
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/60—Medicinal preparations containing antigens or antibodies characteristics by the carrier linked to the antigen
- A61K2039/6031—Proteins
- A61K2039/6037—Bacterial toxins, e.g. diphteria toxoid [DT], tetanus toxoid [TT]
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- 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
- C12N2740/00—Reverse transcribing RNA viruses
- C12N2740/00011—Details
- C12N2740/10011—Retroviridae
- C12N2740/16011—Human Immunodeficiency Virus, HIV
- C12N2740/16211—Human Immunodeficiency Virus, HIV concerning HIV gagpol
- C12N2740/16222—New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
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- 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
- C12N2740/00—Reverse transcribing RNA viruses
- C12N2740/00011—Details
- C12N2740/10011—Retroviridae
- C12N2740/16011—Human Immunodeficiency Virus, HIV
- C12N2740/16211—Human Immunodeficiency Virus, HIV concerning HIV gagpol
- C12N2740/16234—Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
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- AIDS & HIV (AREA)
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- Gastroenterology & Hepatology (AREA)
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Abstract
Description
La demande de brevet principal concerne un produit utile pour la détermination d'un antigène particulier, ledit produit comportant une particule sur iaquelle est fixé au moins un anticorps spécifique dudit antigène particulier. The main patent application relates to a product useful for the determination of a particular antigen, said product comprising a particle to which it is attached at least one antibody specific for said particular antigen.
La présente invention concerne plus particulièrement certains antigènes particuliers qui ont pu etre mis en évidence au moyen du produit décrit dans la demande de brevet principal ; la présente addition concerne en effet les antigènes constitués de protéines non immunogènes du fait de leur poids moléculaire trop faible. The present invention relates more particularly to certain particular antigens which have been able to be demonstrated by means of the product described in the main patent application; the present addition in fact relates to antigens made up of non-immunogenic proteins due to their too low molecular weight.
On s'est aperçu, lors d'essais pharmacodynamiques, que des protéines élaborées à partir de virus du sida ou d'autres maladies notamment tropicales, n'étaient pas efficaces, à savoir, n'engendraient pas la formation d'anticorps neutralisant l'attaque virale. It has been found, in pharmacodynamic tests, that proteins made from AIDS viruses or other diseases, especially tropical, are not effective, that is, do not cause the formation of antibodies neutralizing l viral attack.
Cette inefficacité peut être attribuée au caractère hapténique des antigènes utilisés pour la préparation des vaccins. This ineffectiveness can be attributed to the haptenic nature of the antigens used for the preparation of vaccines.
C'est pourquoi la présente invention concerne l'utilisation d'un ou de plusieurs antigènes pour la préparation de vaccins, caractérisée en ce que le ou les antigènes sont des protéines non immunogènes du fait de leur poids moléculaire trop faible. This is why the present invention relates to the use of one or more antigens for the preparation of vaccines, characterized in that the antigen (s) are non-immunogenic proteins due to their too low molecular weight.
Divers antigènes peuvent être utilisés et seront le plus souvent isolés à partir du sérum de malades. Various antigens can be used and will most often be isolated from the serum of patients.
C'est ainsi que des petites molécules antigéniques ont été isolées par chromatographie d'affinité à partir du sérum de malades atteints du sida (on utilise pour cela les immunoglobulines G (Ig G) d'un patient séro-positif que l'on a préalablement fixées sur du sépharose aminé selon le procédé décrit dans le brevet principal. This is how small antigenic molecules were isolated by affinity chromatography from the serum of patients suffering from AIDS (for this we use the immunoglobulins G (Ig G) of a seropositive patient who has previously fixed on amino sepharose according to the process described in the main patent.
On a notamment isolé la protéine p24, à savoir la protéine de noyau ("core") GAG du retrovirus responsable de syndrome d'immuno-déficience acquise (SIDA). In particular, the p24 protein, namely the GAG core protein, was isolated from the retrovirus responsible for acquired immunodeficiency syndrome (AIDS).
Cette protéine P24 est connue et a déjà fait l'objet de nombreuses publications (Proceedings of the
National Academy of Sciences of the USA 82 (1985) Nov.,
No. 22, Washington, DC, USA).This protein P24 is known and has already been the subject of numerous publications (Proceedings of the
National Academy of Sciences of the USA 82 (1985) Nov.,
No. 22, Washington, DC, USA).
Mais d'autres protéines, non immunogènes, peuvent être utilisées dans le cadre de la présente invention. However, other non-immunogenic proteins can be used in the context of the present invention.
Pour être utilisées selon la présente invention, les protéines doivent être rendues immunogènes par fixation sur un support qui, de préférence, est choisi parmi les molécules permettant de générer les immunités à médiation cellulaire et/ou les immunités à médiation humorale. To be used according to the present invention, the proteins must be made immunogenic by attachment to a support which, preferably, is chosen from molecules making it possible to generate cell-mediated immunities and / or humoral mediated immunities.
Un support tout particulièrement préféré dans le cadre de la présente invention est constitué par une anatoxine par exemple l'anatoxine tétanique. A very particularly preferred support in the context of the present invention consists of a toxoid, for example tetanus toxoid.
Un avantage supplémentaire obtenu lors de l'utilisation de l'anatoxine tétanique est que le vaccin obtenu pourra être utilisé en outre pour prévenir le tétanos. An additional advantage obtained when using tetanus toxoid is that the vaccine obtained can be used in addition to prevent tetanus.
La fixation de la ou des protéines antigéniques sur un tel support peut être effectuée par tous les moyens connus de l'homme de l'art. Elle sera, de préférence effectuée par l'intermédiaire d'un couplage, notamment à l'aide d'une molécule bifonctionnelle telle que le glutaraldéhyde. The fixing of the antigenic protein (s) on such a support can be carried out by any means known to those skilled in the art. It will preferably be carried out by means of coupling, in particular using a bifunctional molecule such as glutaraldehyde.
C'est ainsi que la protéine P24 isolée à partir de malades atteints du sida a été concentrée à 4,6 mg/ml. Thus, the P24 protein isolated from AIDS patients was concentrated to 4.6 mg / ml.
A 1 millilitre de cette concentration, on a ajouté 50 microlitres de glutaraldéhyde à 1 % dans l'eau, puis un-millilitre d'anatoxine tétanique concentrée à 4,6 mg/ml. To 1 milliliter of this concentration, 50 microliters of 1% glutaraldehyde in water were added, then one-milliliter of tetanus toxoid concentrated to 4.6 mg / ml.
Après incubation pendant environ 2 heures à température ambiante et dialyse contre une solution de PBS à pH 7,4, on obtient une composition pharmaceutique. After incubation for about 2 hours at room temperature and dialysis against a PBS solution at pH 7.4, a pharmaceutical composition is obtained.
De façon étonnante, cette composition pharmaceutique s'est révelée être, non seulement efficace à titre préventif (il s'agit alors d'un vaccin) mais aussi à titre curatif. Surprisingly, this pharmaceutical composition has proven to be not only effective as a preventative (it is then a vaccine) but also as a curative.
Dans le cadre de l'immunisation, on a, par exemple, utilisé 0,5 ml de la composition pharmaceutique telle qu'elle a été préparée ci-dessus. As part of the immunization, for example, 0.5 ml of the pharmaceutical composition as used above was used.
On l'a mélangée à 7 ml de phosphate d'alumine et on a injecté le tout par voie-IM chez le mouton. It was mixed with 7 ml of alumina phosphate and the whole was injected by IM route in sheep.
On réalise une piqûre hebdomadaire et on effectue une saignée toutes les deux semaines. A weekly injection is carried out and a bleeding is carried out every two weeks.
Après deux piqûres, c'est-à-dire au quatorzième jour, le mouton présente déjà des anticorps précipitants (réaction d'Ouchterlocy)et agglutination de latex recouvert du virus HIV ou de protéines virales obtenues à partir de surnageant de culture de cellules infectées par le HIV). After two bites, that is to say on the fourteenth day, the sheep already has precipitating antibodies (Ouchterlocy reaction) and agglutination of latex coated with HIV virus or viral proteins obtained from culture supernatant of infected cells by HIV).
Après la quatrième piqûre (saignée au jour 28), les anticorps présentent une très grande affinité pour le virus HIV et les antigènes viraux solubles (réaction d'agglutination au latex). After the fourth injection (bleeding on day 28), the antibodies show a very great affinity for the HIV virus and the soluble viral antigens (agglutination reaction with latex).
Six semaines après le début de l'immunisation, le mouton produit des anticorps neutralisanbs
Dans le but d'illustrer la présente invention, les exemples suivants présentent une partie des essais réalisés lors de l'élaboration à partir de la protéine P24 du vaccin contre le sida. Six weeks after the start of immunization, sheep produce neutralized antibodies
For the purpose of illustrating the present invention, the following examples present a part of the tests carried out during the preparation from the P24 protein of the AIDS vaccine.
EXEMPLE 1
Isolement d'antigènes purifiés :
- Chromatographie d'affinité
Du facteur rhumatoide, isolé par précipitation des euglobulines, a été fixé sur du sépharose 4B aminé par la glutaraldéhyde.EXAMPLE 1
Isolation of purified antigens:
- Affinity chromatography
Rheumatoid factor, isolated by precipitation of euglobulins, was fixed on Sepharose 4B aminated with glutaraldehyde.
Ce réactif a été incubé avec des sérums de patients atteints de SIDA contenant des complexes immuns solubles. This reagent was incubated with sera from AIDS patients containing soluble immune complexes.
Après lavage jusqu'à ce que les liquides de lavage ne contiennent plus de protéines (D.O280 nm = O), on élue avec du tampon glycine - HCL 0,1 M, pH 2,8. After washing until the washing liquids no longer contain proteins (D.O280 nm = O), elution is carried out with glycine buffer - 0.1 M HCL, pH 2.8.
- Fractionnement par chromatographie sur gel
L'éluat concentré a été fractionné par du Sephadex
G200 sur colonne équilibrée par le même tampon que celui de l'élution (glycine-HCL 0,1 M pH 2,8).- Fractionation by gel chromatography
The concentrated eluate was fractionated by Sephadex
G200 on column balanced by the same buffer as that of the elution (glycine-HCL 0.1 M pH 2.8).
Par lecture à 280 nom en U.V. on obtient deux pics. By reading at 280 U.V. names we get two peaks.
Après dialyse de chaque pic contre du NaCl 0,9 % et concentration, on recherche la présence d'Ig G humaines par contre-électrophorèse et par ouchterlony. After dialysis of each peak against 0.9% NaCl and concentration, the presence of human Ig G is sought by counter-electrophoresis and by ouchterlony.
Le pic ne contenant pas d'Ig G est mélangé à de l'adjuvant complet de Freund 0,5/0,5 ml et injecté à un mouton à raison d'une piqûre par semaine. The peak not containing Ig G is mixed with 0.5 / 0.5 ml complete Freund's adjuvant and injected into a sheep at the rate of one bite per week.
Après plus de 6 mois d'observation, le mouton n'a pas produit d'anticorps précipitants par réaction un gel (Ouchterlony) et le mouton est mort en état de cachexie faisant penser au SIDA humain. After more than 6 months of observation, the sheep did not produce precipitating antibodies by reacting a gel (Ouchterlony) and the sheep died in a state of cachexia reminiscent of human AIDS.
EXEMPLE 2
Isolement de complexes antigène - anticorps
par cryoprecipitation. EXAMPLE 2
Isolation of antigen - antibody complexes
by cryoprecipitation.
Des sérums de malades atteints de SIDA ont été laissés pendant au moins 4 jours à 40C. Il se forme un précipité visible à l'oeil nu. Après centrifugation à froid, on recueille le précipité, on le lave 3 à 4 fois avec du
NaCl froid (40C). Ce précipité est resuspendu dans du NaCl froid. Il est mélangé à de l'adjuvant complet de Freund (cf. supra) et injecté à un mouton.Sera from patients with AIDS were left for at least 4 days at 40C. A precipitate is formed which is visible to the naked eye. After cold centrifugation, the precipitate is collected and washed 3 to 4 times with
Cold NaCl (40C). This precipitate is resuspended in cold NaCl. It is mixed with complete Freund's adjuvant (see above) and injected into a sheep.
Ce mouton n'a pas non plus produit d'anticorps précipitants et il est mort en état de cachexie avec des manifestations cliniques faisant penser au SIDA. This sheep also did not produce precipitating antibodies and died in a state of cachexia with clinical manifestations reminiscent of AIDS.
REMARQUES
Pour vérifier que le précipité à froid était constitué de complexes immuns solubles, on l'a dialyse contre un tampon acide (cf., supra) et on observe une redissolution. NOTES
To verify that the cold precipitate was made up of soluble immune complexes, it was dialyzed against an acid buffer (cf., above) and a redissolution is observed.
EXEMPLE 3 Immunisation par des comElexes immuns rendus
insolubles
On insolubilise des Ig G anti HIV sur-des particules de latex (réaction d'agglutination - Cf demande de brevet principal).EXAMPLE 3 Immunization with Rendered Immune ComElexes
insoluble
Anti-HIV Ig Gs are insolubilized on latex particles (agglutination reaction - see main patent application).
Les agglutinats obtenus lors des tests de divers sérums sont recueillis et lavés plusieurs fois avec du
NaCl 0,9 %, puis mélangés à l'adjuvant complet de Freund et injectés à un mouton. Au bout de 4 semaines, on note que le mouton a produit des anticorps anti-HIV et anti-Ig G humaines précipitant en Ouchterlony. Six mois après la dernière injection, le test des anticorps neutralisants était positif. The agglutinates obtained during the tests of various sera are collected and washed several times with
0.9% NaCl, then mixed with Freund's complete adjuvant and injected into a sheep. After 4 weeks, it is noted that the sheep has produced anti-HIV and anti-human Ig G antibodies precipitating in Ouchterlony. Six months after the last injection, the neutralizing antibody test was positive.
REMARQUES
L'analyse par Western Blot du contenu antigénique des agglutinats a montré qu'il s'agissait essentiellement de la protéine p24 et d'autres protéines plus petites du "Core" du virus, donc ce sont des protéines de faibles poids moléculaires qui constituent les antigènes des complexes immuns.NOTES
Western blot analysis of the antigen content of the agglutinates has shown that it is essentially the p24 protein and other smaller proteins of the "Core" of the virus, therefore it is low molecular weight proteins which constitute the immune complex antigens.
Vu leurs petits poids moléculaires ces protéines sont peu immunogènes et de ce fait elles induisent la production d'anticorps de faible affinité. Due to their small molecular weights, these proteins are not very immunogenic and therefore induce the production of low affinity antibodies.
Pour vaincre cet effet hapténique il faut alors les coupler à un support ou leur conférer un haut poids moléculaire par l'insolubilisation (voir supra). To overcome this haptenic effect, it is then necessary to couple them to a support or give them a high molecular weight by insolubilization (see above).
EXEMPLE 4 Couplage~des~Prote~nes a un ~support
Par la méthode de la glutaraldéhyde, des protéines éluées par chromatographie d'affinité (anticorps anti-HIV, couplés au sépharose 4B aminé) sont élués par le thiocyanate d'ammonium 5M après un lavage intensif. (DO250 nm = 0).EXAMPLE 4 Coupling of Proteins to a Support
By the glutaraldehyde method, proteins eluted by affinity chromatography (anti-HIV antibodies, coupled to amino 4B sepharose) are eluted with 5M ammonium thiocyanate after intensive washing. (DO250 nm = 0).
Après dialyse contre le NaCl 0,9 %, l,éluat est concentré jusqu'à 4,6 mg /ml puis couplé à l'anatoxine tétanique concentré à 4,6 mg/ml par de la glutaraldéhyde 1 % dans l'eau après une incubation pendant 2 heures. After dialysis against 0.9% NaCl, the eluate is concentrated to 4.6 mg / ml and then coupled to tetanus toxoid concentrated to 4.6 mg / ml with 1% glutaraldehyde in water after incubation for 2 hours.
Un mouton est ensuite immunisé hebdomadairement par 0,5 ml de ce réactif mélangé à 0,5 ml de phosphate d'aluminium. A sheep is then immunized weekly with 0.5 ml of this reagent mixed with 0.5 ml of aluminum phosphate.
Après 4 semaines d'immunisation le mouton a produit des anticorps précipitants, mais non neutralisants, mais un test effectué deux semaines plus tard c'est-à-dire six semaines après le début de l'immunisation a montré la présence d'anticorps neutralisants. After 4 weeks of immunization the sheep produced precipitating, but non-neutralizing antibodies, but a test carried out two weeks later, that is to say six weeks after the start of immunization, showed the presence of neutralizing antibodies. .
REMARQUES
Un mouton non traité n'a pas produit d'anticorps, de même un mouton auquel on a injecté des antigènes trypanasomiaux, n'a pas produit d'anticorps.NOTES
An untreated sheep did not produce antibodies, nor did a sheep injected with trypanasomial antigens produce antibodies.
Un mouton ayant reçu du virus entier n'a pas, non plus, produit d'anticorps neutralisants. A sheep that received whole virus did not produce neutralizing antibodies either.
EXPERIENCE DE PROVOCATION. PROVOCATION EXPERIENCE.
On a injecté le virus entier à un mouton immunisé par les complexes immuns insolubilisés. A sheep immunized with insolubilized immune complexes was injected with the whole virus.
Après trois mois d'observation il ne présente pas de signe clinique de maladie alors qu'un autre mouton soumis à la même expérience, mais non préalablement immunisé est mort après trois mois et demi et manifestait déjà des signes cliniques après deux mois. After three months of observation, he showed no clinical sign of disease, while another sheep subjected to the same experience, but not previously immunized, died after three and a half months and already showed clinical signs after two months.
Claims (11)
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| FR8718181A FR2625100A2 (en) | 1986-03-13 | 1987-12-24 | Use of antigens for the preparation of vaccines, protein rendered immunogenic and pharmaceutical composition containing it |
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| FR8603572A FR2595826B1 (en) | 1986-03-13 | 1986-03-13 | IMMUNOASSAY PRODUCT, PREPARATION METHOD THEREOF, USE THEREOF, IMMUNOGENIC COMPLEX COMPRISING THE SAME, AND USE OF THE COMPLEX |
| FR8718181A FR2625100A2 (en) | 1986-03-13 | 1987-12-24 | Use of antigens for the preparation of vaccines, protein rendered immunogenic and pharmaceutical composition containing it |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| FR2625100A2 true FR2625100A2 (en) | 1989-06-30 |
Family
ID=26225083
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| FR8718181A Pending FR2625100A2 (en) | 1986-03-13 | 1987-12-24 | Use of antigens for the preparation of vaccines, protein rendered immunogenic and pharmaceutical composition containing it |
Country Status (1)
| Country | Link |
|---|---|
| FR (1) | FR2625100A2 (en) |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1986006414A1 (en) * | 1985-04-29 | 1986-11-06 | Genetic Systems Corporation | Synthetic antigens for the detection of aids-related disease |
| WO1987002775A1 (en) * | 1985-10-24 | 1987-05-07 | Southwest Foundation For Biomedical Research | Synthetic peptides and use for diagnosis and vaccination for aids and arc |
| WO1987002892A1 (en) * | 1985-11-14 | 1987-05-21 | President And Fellows Of Harvard College | T-lymphotrophic virus |
| EP0230222A1 (en) * | 1986-01-06 | 1987-07-29 | F. Hoffmann-La Roche Ag | Expression of HTLV-III gag-Gene |
| EP0241239A2 (en) * | 1986-04-07 | 1987-10-14 | THE UNITED STATES OF AMERICA as represented by the Secretary United States Department of Commerce | A cell line producing aids viral antigens without producing infectious virus particles |
| EP0187041B1 (en) * | 1984-12-24 | 1996-05-15 | Genentech, Inc. | Fusions of AIDS-related polypeptides |
-
1987
- 1987-12-24 FR FR8718181A patent/FR2625100A2/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0187041B1 (en) * | 1984-12-24 | 1996-05-15 | Genentech, Inc. | Fusions of AIDS-related polypeptides |
| WO1986006414A1 (en) * | 1985-04-29 | 1986-11-06 | Genetic Systems Corporation | Synthetic antigens for the detection of aids-related disease |
| WO1987002775A1 (en) * | 1985-10-24 | 1987-05-07 | Southwest Foundation For Biomedical Research | Synthetic peptides and use for diagnosis and vaccination for aids and arc |
| WO1987002892A1 (en) * | 1985-11-14 | 1987-05-21 | President And Fellows Of Harvard College | T-lymphotrophic virus |
| EP0230222A1 (en) * | 1986-01-06 | 1987-07-29 | F. Hoffmann-La Roche Ag | Expression of HTLV-III gag-Gene |
| EP0241239A2 (en) * | 1986-04-07 | 1987-10-14 | THE UNITED STATES OF AMERICA as represented by the Secretary United States Department of Commerce | A cell line producing aids viral antigens without producing infectious virus particles |
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