WO1994001130A1 - Antigene du complexe majeur d'histocompatibilite utilise comme vaccin contre un virus d'immuno-deficience - Google Patents

Antigene du complexe majeur d'histocompatibilite utilise comme vaccin contre un virus d'immuno-deficience Download PDF

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Publication number
WO1994001130A1
WO1994001130A1 PCT/GB1993/001459 GB9301459W WO9401130A1 WO 1994001130 A1 WO1994001130 A1 WO 1994001130A1 GB 9301459 W GB9301459 W GB 9301459W WO 9401130 A1 WO9401130 A1 WO 9401130A1
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WIPO (PCT)
Prior art keywords
antigen
class
cells
human
virus
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Application number
PCT/GB1993/001459
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English (en)
Inventor
Edward James Stott
Peter Anthony KITCHIN
Kingston Henry Gordon Mills
Woon Ling Chan
Mark Page
Lesley Frank TAFFS
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Medical Research Council
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Publication date
Application filed by Medical Research Council filed Critical Medical Research Council
Priority to JP6503132A priority Critical patent/JPH07508984A/ja
Priority to EP93916056A priority patent/EP0650369A1/fr
Priority to AU45762/93A priority patent/AU4576293A/en
Publication of WO1994001130A1 publication Critical patent/WO1994001130A1/fr

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Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
    • C07K14/70503—Immunoglobulin superfamily
    • C07K14/70539—MHC-molecules, e.g. HLA-molecules
    • 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
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies

Definitions

  • VACCINES This invention relates to vaccines against immunodeficiency viruses.
  • Cyno olgus macaques vaccinated with either inactivated partially purified simian immunodeficiency virus (SIV) , fixed SIV-infected C8166 (a human T lymphoblastoid cell line) cells or fixed uninfected C8166 cells can be protected against a challenge infection with the 32H isolate of SIVmac 251 (grown in C8166) 1 ' 2 . Protection is correlated with the levels of antibody response to cellular antigens in the human cells from which the virus immunogen was grown 2"6 . However, the mechanism of protection is unclear.
  • SIV simian immunodeficiency virus
  • the invention provides a major histocompatibility complex (MHC) class I antigen for use in a method of treatment of the human or animal body by therapy, in particular for use as a vaccine against an immunodeficiency virus.
  • MHC major histocompatibility complex
  • the invention also provides a pharmaceutical composition comprising a pharmaceutically acceptable rcarrier or diluent and, as active ingredient, a MHC class I antigen.
  • a pharmaceutical composition comprising a pharmaceutically acceptable rcarrier or diluent and, as active ingredient, a MHC class I antigen.
  • the invention further provides use of a MHC class
  • I antigen in the manufacture of a medicament for use as a vaccine against an immunodeficiency virus I antigen in the manufacture of a medicament for use as a vaccine against an immunodeficiency virus.
  • the antigen is preferably a human class I antigen.
  • the class I molecule may therefore be HLA-A, HLA- B or HLA-C or /3 2 -microglobulin (0 2 m) which is the 0-chain of the HLA class I molecule.
  • the antigen may be the entire class I molecule in which the heavy chain is HLA-A, HLA-B or HLA-C. These are known antigens and can be obtained in purified form. They may be prepared as recombinant proteins.
  • the class I antigen may be given presented by transfected cells, i.e. by cells transfected with a gene encoding the antigen and which consequently express the antigen.
  • Transfected cells which may be administered to a human may be transfected cells of a human diploid cell line. Such cell lines have been tested for safety for the purpose of human vaccine manufacture. An appropriate cell line is the MRC5 cell line.
  • Allogenic lymphocytes which present a class I antigen may be administered to a patient.
  • the lymphocytes may be given as live cells, for example as a blood transfusion. Alternatively they may also be given as fixed or inactivated cells.
  • the lymphocytes may be ones in which the expression of the class I antigen has been enhanced, for example by stimulation with a mitogen or gamma- interferon.
  • the antigen may be used to vaccinate a host against an immunodeficiency virus.
  • the host may be a human or animal but typically it will be wished to vaccinate a human against a human immunodeficiency virus (HIV) .
  • That virus may be HIV-1 or HIV-2.
  • a prophylactic treatment for disease states attributable to infection by an immunodeficiency virus can therefore be provided.
  • the class I antigen may in particular act as an AIDS vaccine.
  • An effective amount of the antigen is administered to a host it is wished to vaccinate.
  • the antigen can be given parenterally, for example subcutaneously or intramuscularly. The amount of antigen per dose depends on a variety of factors such as the age and the condition of the subject involved.
  • a parenteral dose typically consists of from 20 ⁇ g to 1 mg of antigen, for example from 50 to 500 ⁇ g of antigen.
  • a number of doses may be given, for example from 2 to 4 doses over a period of up to six months. Each dose may be given one or two months apart.
  • a pharmaceutical composition also comprising a pharmaceutically acceptable carrier or diluent can be formulated.
  • the composition is thus sterile and pyrogen- free.
  • the composition may also comprise an adjuvant such as Al(OH) 3 or saponin.
  • compositions for intramuscular or subcutaneous injections may contain together with the antigen a pharmaceutically acceptable carrier, e.g. sterile water, olive oil, ethyl oleate, glycols e.g. propylene glycol, and if desired, a suitable amount of lidocaine hydrochloride.
  • a pharmaceutically acceptable carrier e.g. sterile water, olive oil, ethyl oleate, glycols e.g. propylene glycol, and if desired, a suitable amount of lidocaine hydrochloride.
  • the MHC class I antigens can be safely used by virtue of their negligible toxicity.
  • Figure le shows the result of flow cytometry analysis.
  • Example 1 Monkeys were vaccinated as follows: 1179-
  • the 32H cognate isolate of SIVmac 251 and the HIV-1 isolate GB8 were grown in C8166 cells and partially purified by gel exclusion chromatography to minimise loss of envelope glycoprotein 9 . Both virus preparations were inactivated with formalin and dialysed into phosphate- buffered saline (PBS) before use.
  • PBS phosphate- buffered saline
  • 50 ⁇ l of SIVmac (2 ⁇ g/ml) or HIV-1 (16 ⁇ g/ml) diluted in 0.1M carbonate buffer pH 9.6 were added to each well of a 96 well microtitre plate (Nunc, Maxi Sorb) and all subsequent steps carried out as previously described 10 .
  • Monoclonal antibodies (mAbs) and rabbit antibody to human immunoglobulin conjugated to horse radish peroxidase (1:100, DAKO) were diluted in PBS containing
  • Monkey PBMC were isolated from defibrinated blood (obtained from monkeys vaccinated with SIV env) by centrifugation on percoll gradients. PBMC at 1 x 10 6 per ml RPMI 1640 containing 10% autologous serum and 2- mercaptoethanol (5 x 10 "5 M) were dispensed at lOO ⁇ l/well into a 96 well flat-bottomed plate (Costar) .
  • the cultures were incubated at 37°C in a humidified C0 2 incubator for 5 days and were pulsed with l ⁇ Ci/well 3 H-thymidine (Amersha , specific activity 25Ci/mmol) during the final 6 hr of incubation. Cultures in triplicates were harvested on an automatic cell harvester (LKB, Sweden) and radioactivity counted in a ⁇ counter (LKB, Betaplate, Sweden) . In the adsorption experiments, 700 ⁇ l of 1:40 diluted monkey sera were incubated with 10 7 P815 or P815- Al cells followed by a second incubation with 10 7 P815 or P815-B27 cells respectively.
  • Human PBMC were isolated from defibrinated blood (obtained from an individual vaccinated 4 times with botulinum toxoid) by centrifugation on Histopaque (SIGMA) gradients.
  • the assay for T cell proliferation is essentially the same as in Table 2 except that Phytohaemagglutinin (PHA, Wellcome Diagnostics, ll ⁇ g/ml) or botulinum toxoid (obtained from Dr. D. Sesardic, 2.5 ⁇ g/ml) or purified OKT3 (5ng/ml) were used to stimulate the T cells.
  • Ascites fluid of mAbs W6/32 and ADP 314 (anti-HIV-1 p55/p24) were also used as control antibodies.
  • Radioim une precipitation of 35 S-methionine labelled C8166 cell lysate with the monkey sera demonstrate that all the sera from protected but not from unprotected monkeys recognise two major protein bands, 12 and 44kDa (Fig. la) . These bands were precipitated by sera from all the protected animals in the three vaccine groups but not by their preim une sera.
  • Blocking experiments were carried out with 35 S-methionine labelled C8166 cell lysate by pre- precipitation with serum from protected monkeys followed by precipitation with mAb W6/32 (specific for a monomorphic determinant on human MHC class I molecule HLA-A, - B, -C) .
  • the lysate was pre-precipitated with rabbit anti-3 2 m (the ⁇ chain of HLA class I molecule) followed by precipitation with serum from protected monkeys.
  • Results shown in Fig. lb demonstrate that the 12 and the 44kDa bands are 3 2 m and the heavy (h) chain of HLA class I molecule respectively.
  • the intensity and mass of the bands around the 44 Da region precipitated by sera from some protected monkeys suggest that other T cell surface proteins with similar molecular weights such as CD28 (44kDa) , CD2 (Til, 50kDa) and actin (44kDa) may also be recognised.
  • the sera from protected monkeys vaccinated with purified SIVmac 251 virus do not contain anti-CD2 (Fig. lc) or anti-CD28 antibodies (data not shown) .
  • the sera from protected monkeys did not precipitate the ⁇ -chain or the 0-chain of the class I molecule of 35 S-methionine labelled.
  • Herpes papio-transformed monkey B lymphoblastoid cell lines (Fig. Id) , suggesting that the anti-human class I antibodies in the sera are directed at polymorphic regions of the human HLA class I molecule.
  • anti-class I antibody may contribute towards the protection against SIV infection by down-regulating T cell activation, which is known to be required for HIV replication 2 ⁇ "3* *. This is consistent with previous reports that anti-class I antibody can prevent HIV replication in human PMBC 20"22 .
  • the precise mechanism by which anti-class I antibodies down regulate T cell activation is unknown. They may act by inhibiting monocyte antigen presentation. Therefore when protected monkeys were challenged with SIV preparations containing human class I antigens, the anti-human class I antibody could bind and effectively block these antigens from reacting with the monkey antigen presenting cells, thereby preventing the dominating xenogenic activation (>10x higher than anti-SIV response. Table 2) of T cells and hence inhibiting virus replication.
  • the anti-human class I antibody does not cross- react with the monkey class I antigens (Fig. Id) , it would not block the monkey class I antigens (present in SIV grown in. monkey cells) from binding to the monkey antigen presenting cells (Table 2) when the vaccinated monkeys were challenged with that virus. Thus there is no down- regulation of allogenic activation and so virus replication is not inhibited. This may explain the finding that vaccinated monkeys were not protected against challenge infection with SIV grown in monkey cells " >"6 .
  • anti-class I antibody can down- regulate T cell activation by acting directly on the class I molecules on T cells 19 .
  • the anti-human class I antibody from protected monkeys would not be expected to down-regulate the activation of monkey T cells induced by specific antigen by this route, because of the lack of cross-reactivity between the antibody and monkey class I antigens on monkey T cells. Since SIV antigen induced activation is at least 10X weaker than xenogenic or allogenic activation (Table 2), the T cell activation so induced may not be sufficient to support adequate virus replication to infect the monkeys. However, the antibody would be expected to strongly inhibit the activation of human T cells to specific antigens or itogens. This is indeed the case in experiments presented in Table 3. The finding suggests that anti-human HLA class I antibodies may be useful in immunotherapy against HIV infection.
  • the anti-class I antibody contributes towards the resistance to SIV infection in CD4* T lymphocytes, it may not have an effect on the infection of monocytes. In any case, the virus is not expected to replicate extensively in these cells, and virus grown in T cells or PHA activated PBMC would only infect the homologous cell type 3S .
  • the antibody titres in the vaccinees increased further to 1/1920 when assayed by flow cytometry on both P815-B27 and C8166 cells.
  • the antibody titre in control animals remained at ⁇ l/30.
  • the vaccinees had antibody titres of 1/120 - 1/480 whereas that of control animals and prebleeds was ⁇ l/30.
  • Sera from the vaccinees but not the control animals were also shown to bind to the SIV virus envelope as demonstrated by immuno-gold labelling under electron microscopy.

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  • Health & Medical Sciences (AREA)
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  • Chemical Kinetics & Catalysis (AREA)
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  • Gastroenterology & Hepatology (AREA)
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Abstract

On utilise comme vaccin contre un virus provoquant une immuno-déficience un antigène de classe I du complexe majeur d'histocompatibilité. Il peut s'agir d'un antigène humain de classe I tel que l'antigène HLA-A, HLA-B ou HLA-C ou de β2 microglobulines. Le virus peut être un virus d'immuno-déficience humaine tel que le VIH-1 ou le VIH-2.
PCT/GB1993/001459 1992-07-13 1993-07-13 Antigene du complexe majeur d'histocompatibilite utilise comme vaccin contre un virus d'immuno-deficience WO1994001130A1 (fr)

Priority Applications (3)

Application Number Priority Date Filing Date Title
JP6503132A JPH07508984A (ja) 1992-07-13 1993-07-13 免疫不全ウイルスに対するワクチンとして用いるｍｈｃ抗原
EP93916056A EP0650369A1 (fr) 1992-07-13 1993-07-13 Antigènes du complexe majeur d'histocompatibilité utilisés comme vaccin contre un virus de l'immunodéficience
AU45762/93A AU4576293A (en) 1992-07-13 1993-07-13 A major histocompatibility complex antigen for use as vaccine against an immunodeficiency virus

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
GB929214871A GB9214871D0 (en) 1992-07-13 1992-07-13 Vaccines
GB9214871.7 1992-07-13

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WO1994001130A1 true WO1994001130A1 (fr) 1994-01-20

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JP (1) JPH07508984A (fr)
AU (1) AU4576293A (fr)
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GB (1) GB9214871D0 (fr)
WO (1) WO1994001130A1 (fr)

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1996004005A1 (fr) * 1994-08-05 1996-02-15 Angela Turiano Compositions pharmaceutiques contenant des molecules de cmh extraites, destinees a la stimulation du systeme immunitaire
FR2735984A1 (fr) * 1995-06-30 1997-01-03 Inst Nat Sante Rech Med Vaccin contre des agents infectieux ayant une phase intracellulaire, composition pour le traitement et la prevention des infections a hiv, anticorps et procede de diagnostic
WO2000018433A2 (fr) * 1998-09-28 2000-04-06 Fondation Mondiale Recherche Et Prevention Sida Utilisation de complexes antigenes de l'enveloppe du vih et d'antigenes hla de classe i comme vaccin du vih
AU2005202644B2 (en) * 1999-06-01 2009-03-05 Janssen Alzheimer Immunotherapy Prevention and treatment of amyloidogenic disease

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1991016924A1 (fr) * 1990-05-10 1991-11-14 The Dana-Farber Cancer Institute Renforcement de l'association de peptides exogenes avec des molecules mhc de la classe i sur des cellules de systems immuns

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1991016924A1 (fr) * 1990-05-10 1991-11-14 The Dana-Farber Cancer Institute Renforcement de l'association de peptides exogenes avec des molecules mhc de la classe i sur des cellules de systems immuns

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
IMMUNOLOGY LETTERS vol. 24, no. 2, May 1990, AMSTERDAM, THE NETHERLANDS pages 127 - 132 D. LEWIS ET AL. 'HLA-DR peptide inhibits HIV-induced syncytia.' *
THE JOURNAL OF EXPERIMENTAL MEDICINE vol. 176, no. 4, 1 October 1992, NEW YORK, USA pages 1203 - 1207 W-L. CHAN ET AL. 'Protection in simian immunodeficiency virus-vaccinated monkeys correlates with anti-HLA class I antibody response.' *

Cited By (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1996004005A1 (fr) * 1994-08-05 1996-02-15 Angela Turiano Compositions pharmaceutiques contenant des molecules de cmh extraites, destinees a la stimulation du systeme immunitaire
FR2735984A1 (fr) * 1995-06-30 1997-01-03 Inst Nat Sante Rech Med Vaccin contre des agents infectieux ayant une phase intracellulaire, composition pour le traitement et la prevention des infections a hiv, anticorps et procede de diagnostic
WO1997002344A2 (fr) * 1995-06-30 1997-01-23 Institut National De La Sante Et De La Recherche Medicale (Inserm) Vaccin contre des agents infectieux, composition pour le traitement et la prevention des infections a hiv
WO1997002344A3 (fr) * 1995-06-30 1997-03-06 Inst Nat Sante Rech Med Vaccin contre des agents infectieux, composition pour le traitement et la prevention des infections a hiv
US6113902A (en) * 1995-06-30 2000-09-05 Institut National De La Santa Et De La Recherche Medicale (Inserm) Immunogenic compositions comprising peptides from β-2-microglobulin
WO2000018433A2 (fr) * 1998-09-28 2000-04-06 Fondation Mondiale Recherche Et Prevention Sida Utilisation de complexes antigenes de l'enveloppe du vih et d'antigenes hla de classe i comme vaccin du vih
EP1000628A1 (fr) * 1998-09-28 2000-05-17 Fondation Mondiale Recherche et Prevention SIDA Utilisation de complexes antigènes de l'enveloppe de VIH et des antigènes HLA classe I comme vaccin contre le VIH
WO2000018433A3 (fr) * 1998-09-28 2000-07-13 Fond Mondiale Rech & Prev Sida Utilisation de complexes antigenes de l'enveloppe du vih et d'antigenes hla de classe i comme vaccin du vih
AU2005202644B2 (en) * 1999-06-01 2009-03-05 Janssen Alzheimer Immunotherapy Prevention and treatment of amyloidogenic disease

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EP0650369A1 (fr) 1995-05-03
AU4576293A (en) 1994-01-31
JPH07508984A (ja) 1995-10-05
CA2140150A1 (fr) 1994-01-20
GB9214871D0 (en) 1992-08-26

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