EP0344211A1 - Anticorps monoclonaux dans des compositions de vaccins - Google Patents

Anticorps monoclonaux dans des compositions de vaccins

Info

Publication number
EP0344211A1
EP0344211A1 EP88902682A EP88902682A EP0344211A1 EP 0344211 A1 EP0344211 A1 EP 0344211A1 EP 88902682 A EP88902682 A EP 88902682A EP 88902682 A EP88902682 A EP 88902682A EP 0344211 A1 EP0344211 A1 EP 0344211A1
Authority
EP
European Patent Office
Prior art keywords
pathogen
antibody
hbsag
ligand
virus
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.)
Ceased
Application number
EP88902682A
Other languages
German (de)
English (en)
Other versions
EP0344211A4 (fr
Inventor
Giuseppe Colucci
Samuel D. Waksal
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
ImClone LLC
Original Assignee
ImClone Systems Inc
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by ImClone Systems Inc filed Critical ImClone Systems Inc
Publication of EP0344211A1 publication Critical patent/EP0344211A1/fr
Publication of EP0344211A4 publication Critical patent/EP0344211A4/fr
Ceased legal-status Critical Current

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/08—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from viruses
    • C07K16/081—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from viruses from DNA viruses
    • C07K16/082—Hepadnaviridae, e.g. hepatitis B virus
    • 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
    • A61K39/395—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/42—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against immunoglobulins
    • C07K16/4208—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against immunoglobulins against an idiotypic determinant on Ig
    • C07K16/4216—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against immunoglobulins against an idiotypic determinant on Ig against anti-viral Ig
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/42—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against immunoglobulins
    • C07K16/4208—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against immunoglobulins against an idiotypic determinant on Ig
    • C07K16/4233—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against immunoglobulins against an idiotypic determinant on Ig against anti-bacterial Ig
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • 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

  • the present invention involves vaccine formulations which provide for protection against infection by pathogens, either by stimulating an active immune response in a vaccinated host or by conferring short-term passive immunity.
  • the active ingredients of the vaccine formulations of the present invention comprise monoclonal antibodies or fragments of monoclonal antibodies which contain the idiotype of the molecule, such as any Fv fragment including the Fab, F(ab')_, or Fab' and the like.
  • the invention is also directed to methods for preparing such vaccine formulations which do not require isolating or handling the pathogen.
  • the immunogen of the vaccines of the invention which stimulate an active immune response against a pathogen comprises a monoclonal antibody or a fragment thereof, which bears the conformational image of a specific receptor of the pathogen.
  • the monoclonal antibody, or fragment thereof bears the conformational image of a ligand which specifically binds to a receptor on the pathogen.
  • the vaccines of the present invention can confer protection to a host without exposing the host to the pathogen.
  • the invention is demonstrated by way of examples in which monoclonal anti-idiotypes which mimic polymeric human serum albumin were used to generate anti-anti-idiotypes which bear the internal image of hepatitis B virus surface antigen and mimic its immunogenicity in vivo.
  • Vaccines are traditionally prepared by rendering viruses harmless without destroying their immunogenicity. This is achieved either by inactivating the infectivity of the virion, or by selecting an avirulent mutant. Inactivated vaccines are "dead” in the sense that the infectivity of the virions has been destroyed, usually by 5 treatment with ormaldehyde. Injection of these "killed" virus particles into a host will then elicit an immune response capable of neutralizing a future infection with live virus. A number of problems are associated with the use of inactivated vaccines. One major concern is the 0 failure to inactivate all the virus particles. However, even when this is accomplished, because the killed viruses do not multiply in their host, the immunity achieved is often short lived and additional immunizations are required. Another major difficulty encountered in using inactivated
  • 15 vaccines lies in producing enough virus in order to prepare a vaccine which provides the necessary quantity of the relevant antigen to promote systemic antibody production in the host.
  • Attenuated viruses have essentially lost their disease producing ability and generally are good immunogens that provide for long lasting immunity because the attenuated virus actually replicates in the host.
  • several problems are associated with live virus
  • Attenuation of a virus is traditionally accomplished by rapid serial passage of the virus in a foreign host to produce an avirulent mutant.
  • Some of the problems associated with the live virus vaccines include genetic instability of the attenuated virus which results in a greater degree of virulence than is desirable, contamination by adventitious virus during the passages in cell culture, interference by wild type virus and heat __ lability of the live virus in the vaccine formulation.
  • An. alternative to inactivated and live virus vaccines, is the use of a subunit vaccine. This involves immunization of the host solely with the relevant immunogenic material of the pathogenic virus.
  • virus encoded glycoproteins of many of the enveloped viruses or the capsid proteins of non-enveloped viruses are capable of eliciting neutralizing antibodies.
  • Subunit vaccines may be prepared by purifying these proteins from the viruses.
  • One advantage of subunit vaccines is that irrelevant virus material, and the genetic information as well as the replication machinery of the virus are excluded. A major difficulty is encountered in producing the purified proteins in immunogenic amounts.
  • the immunological response to immunogens used in the killed virus vaccines or subunit vaccines can be greatly enhanced if they are administered in an emulsion with adjuvants.
  • the mechanism by which adjuvants increase an immune response are complex, involving the stimulation of activities associated with the reticuloendothelial system.
  • Passive immunization is generally regarded as an emergency procedure for the immediate protection of unimmunized individuals exposed to special risks.
  • passive immunization is also regarded as an important 5 prophylactic measure in several viral infections.
  • human immunoglobulin has proven effective in the short-term prophylaxis of measles and hepatitis A; by contrast, prevention of hepatitis B using human normal immunoglobulin has not been so successful.
  • Anti-idiotypic antibodies or anti-idiotypes are antibodies directed against the antigen-combining region or variable region (called the idiotype) of another antibody molecule.
  • the idiotype of another antibody molecule.
  • Jerne's network model of idiotypic relationships Jerne, N.K., 1974, Ann. Immunol.
  • antigen-combining site for a given antigen should produce a group of anti-antibodies, some of which share with the antigen a complementary structure to the paratope. 35 Immunization with a subpopulation of the anti-idiotypic antibodies should in turn produce a subpopulation of anti- idiotypic antibodies that bind the initial antigen. Thus certain anti-idiotypes directed against a virus-neutralizing antibody should mimic the virus, and when inoculated into a host should induce a specific antiviral response.
  • Vaccines for reovirus have been investigated in experimental animals.
  • Anti-idiotypic antibodies have been used to induce immunity comprising protection against a lethal Sendai virus infection in mice (Ertl, H.C.J. and Finberg, R.W., 1984, Proc. Natl. Acad. Sci. USA 81:2850- 2854) .
  • Immunization with anti-idiotypic antibodies in the HBV system led to the induction of an anti-HBsAg response (Kennedy, supra) .
  • Hepatitis B virus is an etiologic agent in both acute and chronic hepatitis, and has been implicated as an etiologic agent in hepatocellular carcinoma (Hoofnagle,
  • HBV displays a tropism for hepatocytes, the mechanism of which is still an enigma (Hanson, et al., 1979, Infection and Immunity, 26: 125-130) .
  • polyHA polymerized human albumin
  • HBV hepatitis B surface antigen
  • the present invention involves vaccine formulations which provide for active or passive immunity against infection by pathogens, including but not limited to viruses, bacteria, parasites, etc.
  • the active component of the vaccines of the present invention comprises a monoclonal antibody or a fragment of the monoclonal antibody which contains the idiotype of the molecule; these fragments include but are not limited to any Fv fragment such as the
  • the vaccine formulations provide for active immunization of a host against a particular pathogen.
  • the immunogen of thi vaccine comprises a monoclonal antibody, or an idiotypic fragment thereof, which bears the conformational image of a specific receptor of the pathogen.
  • the pathogen is not utilized in the production of the monoclonal antibody.
  • the monoclonal antibodies which mimic a specific receptor of the pathogen may comprise: (a) a monoclonal antibody generated against a ligand that binds specifically to the receptor of the pathogen; (b) a monoclonal antibody generated against the idiotype of a second antibody which, in turn, defines the idiotype of a
  • the vaccine formulations may provide for passive immunity to confer short-term resistance to infection by a pathogen.
  • the active ingredient of the vaccine comprises a monoclonal antibody or idiotypic fragment thereof which bears the conformational image of a ligand that binds specifically to a receptor of the pathogen.
  • the pathogen is
  • the monoclonal antibodies which mimic a ligand that binds to a specific receptor of the pathogen may comprise: (a) a monoclonal antibody directed against the idiotype of a second antibody which, in turn, is directed against the
  • ___ ligand (b) a monoclonal antibody directed against the 2b idiotype of a second antibody which, in turn, is directed against the idiotype of a third antibody which, in turn, is directed against the idiotype of a fourth antibody which is directed against the ligand; or (c) any other anti-idiotypic antibody which mimics the ligand that binds to the pathogen as depicted in the cascade shown in FIG. 1. In all cases the monoclonal antibody should also competitively inhibit the binding of the ligand to the pathogen.
  • Some of the advantages of the vaccine formulations of the present invention include the following: (a) a pathogenic immunogen is not required to stimulate the immunity of the host;
  • the vaccines of the present invention can be prepared against epitopes that comprise non-proteins such as lipids, carbohydrates, or glycolipids, etc.;
  • Anti-id anti-idiotype antibody
  • BSA bovine serum albumin
  • Fv the variable region or antigen-combining site of an antibody molecule. This may be any fragment which contains the idiotype of the molecule including but not limited to the Fab, F(ab') 2 , Fab', and the like.
  • HA human albumin (monomeric)
  • HBV hepatitis B virus
  • HBsAg hepatitis B surface antigen
  • PEG polyethylene glycol
  • polyHA polymeric human albumin
  • SDS sodium dodecylsulfate
  • TWEEN-20 polyoxyethylene sorbitan onolaurate
  • FIG. 1 is a diagrammatic representation of the cascade 5 of anti-idiotypic antibodies that can be generated from a ligand which specifically binds to a receptor of a pathogen.
  • the antibodies that can be used as the active ingredient in either the active or passive vaccine formulations of the present invention are indicated in the figure.
  • the anti ⁇ ng ligand antibody and anti-id2, anti-id4, etc. mimic the receptor of the pathogen and can be used as the immunogen in the vaccines formulated for active immunization.
  • the anti- idl, anti-id3, etc. mimic the ligand which binds specifically to the receptor of the pathogen, and can be 15 used as the active ingredient in the vaccines formulated for passive immunization.
  • the vaccine formulations of the invention can provide for protection against infection of a host by a number of pathogens, including but not limited to viruses, bacteria, parasites, etc.
  • the active ingredient of the vaccine formulations of the present invention comprise monoclonal antibodies or fragments of the monoclonal antibody which contain the idiotypic region of the antibody molecule; these include, but are not limited to the fragments which include the Fv region, such as the Fab, F(ab')_, Fab' fragments and the like.
  • the monoclonal antibodies of the present invention are generated without utilizing the pathogen.
  • antibodies, or fragments thereof are formulated as a vaccine which, depending upon the nature of the idiotype, can be used either to stimulate an active immune response or to confer short-term passive immunity in a host against a pathogen.
  • the vaccine formulations of the present invention which provide for either active or passive immunity are described in separate sections below, and with reference to FIG. 1 which diagrammatically represents the cascade of anti-idiotypic antibodies that could be used in different embodiments of the invention.
  • receptors on their surfaces that specifically bind to particular substrates or receptors
  • ligands that are present in the host which is the target for infection by the pathogen. 5
  • the binding interaction between the pathogen receptor and the host's ligand enables the attachment of the pathogen to a target host cell surface. This attachment is generally a prerequisite for successful infection, multiplication, colonization, etc. by the pathogen which
  • the host ligand may be used to generate an antibody (or its anti-anti—idiotype) which mimics the pathogen receptor (see
  • This antibody may be used in vaccine formulations to induce active immunity against the pathogen.
  • anti-pathogen antibody obtained from a host who has developed a neutralizing immune response (e.g. , through exposure to the pathogen) may be used as the starting material to develop the anti-idiotype cascade.
  • the anti-idiotype of the neutralizing antibody which mimics the conformational structure of the receptor of the pathogen and induces a neutralizing response is selected as the active ingredient of the vaccine formulation.
  • a vaccine can be formulated without using or handling the pathogen.
  • the vaccine formulations and their method of preparation in accordance with the invention is based, in part, upon the theory that the idiotypic region of an antibody molecule directed against the idiotype of a neutralizing antibody or a ligand which binds specifically to a receptor on a pathogen should bear the conformational image and structure of the receptor on the pathogen; for example, see anti-ligand in FIG. 1.
  • an antibody directed against the idiotype of a second antibody which, in turn, is directed against the idiotype of the anti-ligand antibody should also bear the conformational image of the receptor on the pathogen; for example, see anti-id2 in FIG.
  • FIG. 1 which diagrammatically depicts the cascade of anti- idiotypes that can be generated from the ligand and/or the anti-ligand.
  • the anti-ligand as well as anti-id2, anti-id4, etc. in the cascade would be expected to mimic the receptor of the pathogen.
  • the degree to which the anti-ligand antibody or anti-idiotypic antibody mimics the receptor on the pathogen can be ascertained in a competitive binding assay. For example, if the anti-ligand, or the anti- idiotype, competitively inhibits the binding of the pathogen to the ligand, then one can conclude that the idiotype of the antibody molecule mimics the receptor on the pathogen.
  • an effective dose of the anti-ligand antibody, or the appropriate anti-idiotypic antibody which mimics the receptor on the pathogen may be formulated as an immunogen in a vaccine which is used to stimulate an active immune response in a vaccinated host. Because the idiotype of the antibody molecule mimics the conformation of the receptor on the pathogen, the resulting immune response directed against the idiotype will also be directed against the receptor on the pathogen. Since this receptor is responsible for the attachment of the pathogen to its target, the immune response should neutralize the infectivity of the pathogen.
  • the anti-ligand antibodies which are used as the immunogen in the vaccine formulations of the present invention should be monoclonal antibodies in order to insure a continuous supply and in order to be able to produce the antibody in large quantities.
  • the intermediate antibodies of the cascade need not be monoclonal antibodies.
  • the anti- ligand, and anti-idl, anti-id2, and anti-id3 need not be monoclonal antibodies.
  • the monoclonal antibodies which are used as immunogens can be prepared using any technique which provides for the production of antibody molecules by continuous cell lines in culture. For example, the hybridoma technique originally developed by Kohler and Milstein (1980, Sci. Am. 243(4):
  • the invention is not limited to the use of whole antibody molecules as the immunogen in the vaccine formulations, rather, any fragment of the antibody which contains the idiotype of the antibody (e.g. , the Fv portion of the antibody molecule) could be used.
  • Such fragments include but are not limited to: the F(ab')_ fragment which can be generated by treating the antibody molecule with pepsin; the Fab' fragments which can be generated by reducing the disulfide bridges of the F(ab')_ fragment; and the 2Fab or Fab fragments which can be generated by treating the antibody molecule with papain and a reducing agent to reduce the disulfide bridges.
  • the F(ab') fragments would be preferred as immunogen, in order to enhance the induction of an immune response of the desired specificity.
  • the pathogens for which the vaccines can be prepared are numerous. For example, in many viruses, attachment of virions to a target cell is a prerequisite for penetration of the virus into the host cell and subsequent multiplication. Specificity of attachment has been demonstrated for viruses such as poliovirus and influenza virus. Poliovirus, for instance, will attach to primate but not to rodent cells. This specificity is believed to be due to the interaction of the virus with a specific cellular receptor of the primate. According to the invention, the receptor can be used as the ligand from which anti-ligand and/or the cascade of anti-idiotypic antibodies could be generated. The appropriate antibodies could be used in a vaccine formulation as described.
  • the presence of certain proteins on the surface of the host cell is required for the attachment of influenza virus.
  • These cell-surface proteins appear to be receptors for the viral hemagglutinin antigen.
  • the receptor proteins can be used as the ligand from which the anti-ligand and/or cascade of anti-idiotypic antibodies are generated.
  • the appropriate antibodies can be formulated in a vaccine as described.
  • Penetration of the host cell during retrovirus infection has been shown to be dependent upon a specific interaction between the viral envelope glycoprotein and a host cellular receptor (Crittenden, L.B., 1968, J. Natl. Cancer Inst. 41: 145-153; Piraino, F., 1967, Virology 32:700-707; Steck, F.T., and Rubin, H. , 1965, Virology :642-653).
  • these host receptors can be used as the immunogen for generation of the anti- idiotypic cascade of antibodies.
  • an anti- idiotypic cascade can be used to produce antibodies to the cellular receptor for the AIDS virus, the etiological agent of Acquired Immune Deficiency Syndrome (AIDS) (Gallo, R.C., et al. , 1984, Science 224:500; Popovic, M. , et al., 1984, Science 224:497; Barre-Sinoussi, F. , et al., 1983, Science 220: 868; Levy, J.A., et al., 1984, Science 225: 840).
  • AIDS Acquired Immune Deficiency Syndrome
  • CD4 T4 antigen on lymphocytes
  • AIDS virus epitopes may be used as the initial antigens for the eventual production of anti-idiotypic antibodies that mimic the viral protein which binds the cellular T4 antigen.
  • _ epitopes are the currently available diagnostic kits for
  • AIDS many of which contain disrupted AIDS virus virions.
  • AIDS virus proteins which are produced by recombinant DNA techniques, chemically synthesized, obtained from purified virus, etc.
  • anti-AIDS virus antibody derived from patients could be used as the starting point for generation of anti-idiotypic antibodies directed against the cellular receptor for the AIDS virus.
  • HBV which has binding sites for polymerized serum albumin, attaches to the surface of hepatocytes via the polymerized albumin; the HBV-polyalbumin complex is then taken into the liver cells by endocytosis.
  • HBV particles and HBsAg particles only bind specifically to human and chimpanzee 5 serum polyalbumin.
  • the polymerized human albumin is used as the ligand from which the anti-ligand and/or anti-idiotypic antibodies are generated.
  • a cascade of anti-idiotypic 5 antibodies can be generated using the human anti-polyHA which can be obtained from patients infected with HBV.
  • the appropriate antibodies can be formulated as a vaccine as described herein.
  • cholera toxin choleragen
  • ganglioside G M _ ganglioside G M _
  • G M1 a host cell ganglioside
  • G M _ may be used as the immunizing ligand from which anti-idiotypic antibodies are generated.
  • This embodiment of the invention is based, in part, upon the theory that the idiotypic region of an antibody 25 molecule directed against the idiotype of a second antibody which, in turn, is directed against a ligand which binds specifically to a receptor on a pathogen should bear the conformational image and structure of the ligand; for
  • anti-idl see anti-idl in FIG. 1.
  • Other anti-idiotypic 30 antibodies such as anti-id3, indicated in the cascade depicted in FIG. 1 would also be expected to mimic the ligand which binds specifically to the receptor on the pathogen.
  • the degree to which the anti-idiotypic antibody mimics the ligand can be ascertained in a competitive wO binding assay. For example, if the anti-idiotypic antibody competitively inhibits the binding of the ligand to the pathogen, then one can conclude that the idiotype of the antibody molecule mimics the ligand.
  • the anti-idiotypic antibody which mimics the ligand is formulated in a "vaccine" that can be used to provide passive immunization for short-term protection against infection by the pathogen. Because the idiotype of the anti-idiotypic antibody mimics the conformation of the ligand, the antibody will bind to the receptor on the pathogen. Binding of the antibody to the receptor will interfere with attachment of the pathogen to its target and, therefore, will prevent subsequent infection by the pathogen.
  • the anti-idiotypic antibodies used in the vaccine formulations for passive immunization should be monoclonal antibodies in order to insure a continuous supply and in order to be able to produce the antibody in large quantities.
  • the intermediate antibodies of the cascade need not be monoclonal antibodies.
  • the intermediate antibodies of the cascade such as anti-id2, anti-idl, and the anti- ligand antibody need not be monoclonal antibodies.
  • the monoclonal antibodies used in the vaccine formulations for passive immunization can be prepared by any technique that provides for the production of antibody molecules by continuous cell lines in culture.
  • the hybridoma technique originally developed by Kohler and Milstein (1980, Sci. Amer. 243(4): 66-74) as well as other techniques which have recently become available such as the human B-cell hybridoma technique (Kozbar et al., 1983, I munology Today 4: 72) and the EBV-hybridoma technique to produce human monoclonal antibodies (Cole et al., 1985, Monclonal Antibodies and Cancer Therapy, Alan R. Liss, Inc. , pp. 77-96) and the like are within the scope of the present 5 invention.
  • Human monoclonal antibodies are preferred for use, since they would reduce the possibility of eliciting an immune response directed against the antibody molecule. Repeated administrations of the monoclonal antibody for 10 passive immunization would preferably be minimized so as to reduce the risk of inducing auto-immunity.
  • the invention is not limited to the use of whole monoclonal anti-idiotypic antibody molecules as the active ingredient in vaccine formulations for passive immunity. 15 Rather, any fragment of the antibody which contains the idiotype of the antibody (e.g. , the Fv portion of the antibody molecule) could be used. Such fragments include but are not limited to: the F(ab')_ fragment which can be generated by treating the antibody molecule with pepsin; the Fab' fragments which can be generated by reducing the
  • disulfide bridges of the F(ab')_ fragments 20 disulfide bridges of the F(ab')_ fragments; and the 2Fab or Fab fragments which can be generated by treating the antibody molecule with papain and a reducing agent to reduce the disulfide bridges.
  • ____.0 one that includes the Fc region of the molecule) as the active ingredient of a vaccine formulation for passive immunity may offer advantages over using an Fv fragment of the antibo n dy.
  • the monoclonal anti-idiotypic antibody is of a class that activates complement or mediates
  • ADCC antibody dependent cellular cytotoxicity
  • Passive immunization can be used to treat a variety of viral or bacterial diseases.
  • Hepatitis A virus has successfully been treated with human immunoglobulin to provide passive immunity.
  • This human immunoglobulin is obtained from donors who have had contact with the virus.
  • monoclonal antibodies directed against envelope proteins of HAV could be used to inactivate the hepatitis A virus via passive immunization.
  • Anti-idiotypic antibodies directed against either the bacterial antigen or cellular receptor can then be used in passive immunization.
  • Bacteria which are intestinal pathogens such as Shigella dysenteriae, Vibrio cholerae, various Salmonella and E. coli species and others can be treated by passive immunization schemes in which the anti-idiotypic antibody is taken orally.
  • the vaccine formulations which stimulate an active immune response can be prepared by mixing the monoclonal antibody, or an Fv fragment thereof, which mimics the receptor or the pathogen in a carrier suitable for use in vivo.
  • the monoclonal antibody or its Fv fragment should be formulated with a suitable adjuvant in order to enhance the i ⁇ imunological response of the host.
  • suitable adjuvants include, but are not limited to aluminum hydroxide, surface active substances, lysolecithin, pluronic polyols, polyanions, and peptides.
  • Other potentially useful adjuvants in humans such as corynebacterium parvum and BCG
  • the monoclonal antibody or the Fv fragment could be incorporated into or on liposomes or conjugated to polysaccharides and/or other polymers that are useful in vaccine formulations.
  • the antibody fragment is a hapten, i.e. , antigenic but not capable of eliciting an immune response, it may be conjugated to a large carrier molecule, such as protein serum albumin and the like to confer immunogenicity to the complex.
  • the vaccine could be formulated as a multivalent vaccine.
  • a mixture of different antibodies, each of which mimics the receptor of a different pathogen can be mixed together in one formulation.
  • PASSIVE IMMUNIZATION The monoclonal antibody molecules which mimic the ligand that binds to the pathogen can be formulated to confer short-term passive immunity to the host.
  • Adjuvants 5 are not needed in this type of preparation because the object of this formulation is not to stimulate an immune response, but rather to bind to and inactivate the pathogen.
  • any suitable pharmaceutical carrier may be used. Passive immunization could be used on an emergency basis for 0 immediate protection of unimmunized individuals exposed to special risks.
  • monoclonal antibody that mimics polymeric human albumin which, according to the present invention, is a ligand that binds to HBV.
  • This monoclonal antibody could be used in a vaccine formulation that provides for passive immunity; alternatively, the monoclonal antibody could be used to
  • polyHA polymeric human albumin
  • Inhibition assays showed that only polyHA (polymeric human albumin) could block the binding activity of antibodies 35 63.14 and 70.F9, as compared to HA (monomeric human albumin) and two polyclonal anti-HBsAg antibodies.
  • a partial inhibition of the anti-HBsAg reactivity of the anti-id by a monoclonal a ⁇ ti-HBsAg is probably due to steric hindrance..' 5
  • These data suggest that the anti-id share a common three dimensional structure with polyHA, and recognize a polyHA binding site expressed on HBsAg. This recognition occurs on a domain of the antigen different from that detected by conventional anti-HBsAg antibodies.
  • the antigen which seems not to be detected by conventional anti-HBsAg antibodies Since the expression of the polyHA receptor on HBV appears to correlate with a higher viral replication and infectivity, our anti-id can be valuable in identifying different subsets of patients with viral 0 hepatitis and the healthy carriers who are more likely to transmit the infection. Finally, if the polyHA receptor is involved in the penetration of HBV into the liver cells these anti-id when used in a passive immunization scheme might be effective in preventing the viral infection. 5 Alternatively, an anti-idiotypic antibody generated against the anti-id described herein could be used in an active vaccine formulation.
  • mice were immunized with an affinity purified human IgG anti-polyHA obtained from a patient with autoimmune hepatitis. They were at first injected subcutaneously in the inguinal and axillary regions and in the footpads with 100 micrograms of the immunogen in complete Freund's adjuvant. The successive boosts were given intraperitoneally using incomplete Freund's adjuvant and saline. After the third injection in saline, mice were bled and the serum tested for the presence of anti-id using a hemagglutination inhibition assay (HIA) . Human red blood cells were coated with polyHA according to S.N. Thung et al.
  • HAA hemagglutination inhibition assay
  • PolyHA was generated by cross-linking with glutaraldehyde according to R. Lenkei
  • Spleen cells obtained from the ⁇ positive mice were fused with the HAT-sensitive cell line SP2/0-AZ-14, using PEG 44%. After two weeks of HAT selection, hybridomas were screened for the presence of anti-id with the HIA, as previously 0 described. The positive clones were expanded, subcloned by limiting dilutions and grown in ascites.
  • Fab' monoclonal anti-id 2 of the immunogen as well as HBsAg was tested in a solid phase RIA, in which 11 different human myeloma proteins and an unrelated monoclonal anti-id (MOPC 173) served as negative control.
  • HBsAg obtained from the concentrated supernatant of the HBV-transfected 3T3 cell
  • ELISA Monoclonal anti-id 63.14 and 70.F9 were conjugated with alkaline phosphatase (type VII-T, Sigma Chemical Corporation, St. Louis, MO) according to Voller et al. (1976, Bull. World Health Organ. 53: 55-65). The wells of a microtiter plate were coated with HBsAg (10 micrograms/ml) , by incubating overnight at 4 ⁇ C. Supernatants obtained from the untransfected 3T3 fibroblasts served as control.
  • alkaline phosphatase type VII-T, Sigma Chemical Corporation, St. Louis, MO
  • HBsAg after preincubation with different dilutions of the unconjugated anti-id.
  • the antibodies affinity constant (k) was calculated from the molar concentration of the inhibitor giving 50% inhibition, the molar concentration of the tracer and the amount of tracer bound in the absence of the inhibitor. 6.1.5.
  • WESTERN BLOT Purified HBsAg obtained from the 4.10 cell supernatant and from the serum of a patient with acute hepatitis, were electrophoresed on a 12% SDS-polyaery1amide gel, overnight 5 at 10 A. The proteins were then electrophoretically transferred to a nitrocellulose filter for 3 hours at 60 V.
  • the filter was then blocked with 5% BSA in 1 mM Tris-HCl buffer, pH 7.6, and incubated with the monoclonal anti-id overnight at 4°C. 0 After washing with Tris-Tween, the filter was incubated with horseradish peroxidase- ⁇ onjugated goat anti-mouse IgG (BioRad Laboratories, Richmond, CA) , washed, and reacted with the peroxidase substrate solution (0.01% 4-chloro-l- napthol in methanol and 0.02% hydrogen peroxide).
  • the 1 monoclonal anti-HBsAg H25B10 and cellular extract obtained from 3T3 fibroblasts served, respectively, as positive and negative controls.
  • the cells were incubated with the avidin-biotin-peroxidase complex (ABC, Vector Laboratories, Burlingame, CA) .
  • the peroxidase reaction was then developed using diaminobenzidine and hydrogen peroxide as substrate.
  • PolyHA and the monoclonal antibody H25B10 served as blocking agents.
  • H25B10 and the monoclonal anti-id MOPC 173 served respectively as positive and negative controls. 5 6.1.7.
  • mice were immunized with purified human IgG specific for human polymeric albumin in order to induce an anti-idiotypic response which would bear the internal image of polyHA and mimic its binding activity as described.
  • Fusions were done using spleen cells derived from mice whose sera were positive in the RIA assay. Six clones derived from these fusions subsequently showed positive reactivity in this assay; four of these hybrido as were then expanded, subcloned and further analyzed for their ability to bind the
  • Binding was assayed in a solid phase RIA.
  • the human proteins tested were immobilized in sample wells which were incubated with one of the mouse anti-ids indicated. Binding of 5 the anti-id was determined by a second incubation with I- radiolabled anti-mouse antibody. The plates were washed and counted in a gamma-counter. Ascites derived from the mouse myeloma cell line MOPC 173 was used as a negative control. Results are expressed as mean and standard deviation of triplicates. 6.2.2. POLY-HA INHIBITS BINDING
  • molecular weight markers we used the following proteins: bovine albumin, ovalbumin, glyceraldehyde-3- phosphate dehydrogenase, carbonic anhydrase, trypsinogen, trypsin inhibitor, alpha-lactalbumin (Sigma Chemical Co., St. Louis, MO) .
  • HBsAg appeared to be specific for the common "a" determinant of HBsAg, since they reacted equally well against two different antigen subtypes and precipitated the 24-kD a protein of the viral particles.
  • HBsAg obtained from the supernatant of the HBV-transfected cell line 4.10 (Christman et al., 1982, Proc. Natl. Acad. Sci. (USA) 79:1815), was purified on a cesium chloride gradient and adjusted to a concentration of 1 ⁇ g/ml in 0.05 M sodium carbonate buffer, p 9.6. The antigen as well as normal human serum were then use to coat the wells of a microtiter plate by incubating overnight at 4 ⁇ C.
  • alkaline phosphatase-conjugated 63.14, HB25B10 American Cell Type Culture Collection, Rockville, MD or polyHA. These molecules were conjugated with alkaline phosphatase (type VII-T, Sigma Chemical Co., St. Louis, MO) according to Voller et al. (1976, Bull WHO 53:55). For inhibition experiments 63.14, HB25B10 and polyHA were preincubated or coincubated with an aliquot of hybridomas supernatant.
  • hybridomas were indeed able t mimic HBsAg, we tested them for the ability to bind a monoclonal anti-HBsAg (H25B10) , as well as polyHA in ELISA.
  • Enzyme-conjugated H25B10 and polyHA were reacted with anti-anti-Id supernatants previously bound to the wells o a plastic 96-well plate. Results are expressed as mean + SD (A 405 nm) of duplicates.
  • Anti-anti-Id, Gil was used in studies testing its abilit to induce an immune response against HBsAg in vivo.
  • rabbits were used as the hosts and were injected with 0.5 mg of Gil or HBsAg in complete Freund's adjuvant and boosted subcutaneously 1 month later. After the second immunization the rabbits' sera were tested in ELISA against two different preparations of HBsAg (adw, ayr) . Serum obtained from rabbits injected with anti-anti-Id was found to express anti-HBsAg activity, with a titer almost as high as the showed by rabbits immunized with HBsAg.

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Organic Chemistry (AREA)
  • Immunology (AREA)
  • Medicinal Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Biophysics (AREA)
  • Genetics & Genomics (AREA)
  • Biochemistry (AREA)
  • Molecular Biology (AREA)
  • Virology (AREA)
  • Microbiology (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Communicable Diseases (AREA)
  • Mycology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Epidemiology (AREA)
  • Animal Behavior & Ethology (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
  • Peptides Or Proteins (AREA)

Abstract

Des compositions de vaccins confèrent une immunité active ou passive contre des infections causées par des pathogènes. Le composant actif des vaccins de la présente invention consiste en un anticorps monoclonal ou un fragment de l'anticorps monoclonal qui contient l'idiotype de la molécule. Des compositions de vaccins qui assurent l'immunisation active d'un hôte contre un pathogène particulier sont préparés en utilisant des anticorps monoclonaux ou des fragments idiotypiques qui portent l'image conformationnelle d'un récepteur spécifique du pathogène. Des compositions de vaccins qui confèrent l'immunité passive sont préparées à partir d'anticorps anti-idiotypiques ou des fragments idiotypiques de ceux-ci qui portent l'image conformationnelle d'un ligand qui se lie de manière spécifique à un récepteur du pathogène. Dans un cas comme dans l'autre, il n'est pas nécessaire que le pathogène prépare l'anticorps qui comprend le composant actif de la composition du vaccin.
EP19880902682 1987-02-20 1988-02-19 Anticorps monoclonaux dans des compositions de vaccins. Ceased EP0344211A4 (fr)

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
US1680787A 1987-02-20 1987-02-20
US16807 1987-02-20
US15669188A 1988-02-17 1988-02-17
US156691 1988-02-17

Publications (2)

Publication Number Publication Date
EP0344211A1 true EP0344211A1 (fr) 1989-12-06
EP0344211A4 EP0344211A4 (fr) 1990-03-12

Family

ID=21779094

Family Applications (1)

Application Number Title Priority Date Filing Date
EP19880902682 Ceased EP0344211A4 (fr) 1987-02-20 1988-02-19 Anticorps monoclonaux dans des compositions de vaccins.

Country Status (4)

Country Link
EP (1) EP0344211A4 (fr)
JP (1) JPH02502183A (fr)
AU (2) AU1484688A (fr)
WO (1) WO1988006040A1 (fr)

Families Citing this family (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5955076A (en) * 1989-06-15 1999-09-21 Peptide Therapeutics Limited Immunoactive peptides and antibodies and their use in anti-allergy treatment
GB8913737D0 (en) * 1989-06-15 1989-08-02 Univ Birmingham A novel anti-allergy treatment
KR100498198B1 (ko) 1996-03-01 2005-09-09 노파르티스 아게 알레르기에 대한 백신접종 및 치료를 위한 펩티드 임뮤노겐

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0142345A2 (fr) * 1983-11-09 1985-05-22 Synbiotics Corporation Vaccin anti-idiotypique à base d'anticorps monoclonaux anti-idiotypiques
WO1986001539A1 (fr) * 1984-08-27 1986-03-13 The Trustees Of Columbia University In The City Of Procede de production d'anticorps monoclonaux anti-idiotypiques
WO1988001304A1 (fr) * 1986-08-21 1988-02-25 The Trustees Of Columbia University In The City Of Adn de codage de la proteine t4 de la surface des cellules t et utilisation de fragments de t4 pour le traitement du sida

Family Cites Families (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4596792A (en) * 1981-09-04 1986-06-24 The Regents Of The University Of California Safe vaccine for hepatitis containing polymerized serum albumin
US4661586A (en) * 1981-11-17 1987-04-28 The Board Of Trustees Of The Leland Stanford Junior University Monoclonal anti-idiotype antibodies
JPS59116230A (ja) * 1982-11-29 1984-07-05 ベイロ−・カレツジ・オヴ・メデイスン ウイルス性感染因子に対する免疫応答を生起する抗−イデイオタイプ抗体
EP0141783B2 (fr) * 1983-11-07 1993-06-16 The Wistar Institute Réponse immune à des tumeurs et des virus induite par des anticorps anti-idiotypes
CA1256795A (fr) * 1983-12-28 1989-07-04 Dennis A. Carson Anticorps anti-idiotype induits par des polypeptides de synthese
US4683295A (en) * 1984-05-24 1987-07-28 Scripps Clinic And Research Foundation Method for the preparation of anti-receptor antibodies

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0142345A2 (fr) * 1983-11-09 1985-05-22 Synbiotics Corporation Vaccin anti-idiotypique à base d'anticorps monoclonaux anti-idiotypiques
WO1986001539A1 (fr) * 1984-08-27 1986-03-13 The Trustees Of Columbia University In The City Of Procede de production d'anticorps monoclonaux anti-idiotypiques
WO1988001304A1 (fr) * 1986-08-21 1988-02-25 The Trustees Of Columbia University In The City Of Adn de codage de la proteine t4 de la surface des cellules t et utilisation de fragments de t4 pour le traitement du sida

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
EUROPEAN JOURNAL OF IMMUNOLOGY, vol. 17, 1987, pages 365-370, Verlagsgesellschaft mbH, Weinheim, DE; G. COLUCCI et al.: "Interactions between hepatitis B virus and polymeric human albumin. I Production of monoclonal anti-idiotypes (anti-anti-polymeric human albumin) which recognize hepatitis B virus surface antigen" *
EUROPEAN JOURNAL OF IMMUNOLOGY, vol. 17, 1987, pages 371-374, Verlagsgesellschaft mbH, Weinheim, DE; G. COLUCCI et al.: "Interactions between hepatitis B virus and polymeric human serum albumin II. Development of syngeneic monoclonal anti-anti-idiotypes which mimic hepatitis B surface antigen in the induction of immune responsiveness" *
See also references of WO8806040A1 *
THE JOURNAL OF IMMUNOLOGY, vol. 137, no. 9, 1st November 1986, pages 2937-2944, The American Association of Immunologists, US; J.S. McDOUGAL et al.: "Binding of the human retrovirus HTLV-III/LAV/ARV/HIV to the CD4 (T4) molecule: conformation dependence, epitope mapping, antibody, inhibition, and potential for idiotypic mimicry" *

Also Published As

Publication number Publication date
EP0344211A4 (fr) 1990-03-12
AU5217890A (en) 1990-09-06
JPH02502183A (ja) 1990-07-19
WO1988006040A1 (fr) 1988-08-25
AU1484688A (en) 1988-09-14

Similar Documents

Publication Publication Date Title
Uytdehaag et al. Induction of neutralizing antibody in mice against poliovirus type II with monoclonal anti-idiotypic antibody.
Reagan et al. Anti-idiotypic antibodies induce neutralizing antibodies to rabies virus glycoprotein
Sharpe et al. Syngeneic monoclonal antiidiotype can induce cellular immunity to reovirus.
Kennedy et al. Common idiotypic determinant associated with human antibodies to hepatitis B surface antigen.
AU596070B2 (en) Anti-idiotype antibodies & their use in inducing immulogical response to viruses & tumours
US5856087A (en) Method for detecting immune response to hepatitis B
PT85137B (pt) Processo de preparacao de um polipeptido de composito imunogenico, de melhoramento da imunogenicidade de um imunogenio e de atenuacao da falta de capacidade de resposta a uma vacina do virus da hepatite b
Kang et al. Anti-idiotype monoclonal antibody elicits broadly neutralizing anti-gp120 antibodies in monkeys.
Carayanniotis et al. Delivery of synthetic peptides by anti-class II MHC monoclonal antibodies induces specific adjuvant-free IgG responses in vivo
Anders et al. Induction of immune response to influenza virus with anti-idiotypic antibodies
US20050163768A1 (en) Monoclonal antibodies and their use
EP0344211A1 (fr) Anticorps monoclonaux dans des compositions de vaccins
JP2006143738A (ja) ルイスｙ−特異的モノクローナル抗体ｂｒ５５−２に対する抗イディオタイプモノクローナル抗体およびその使用
EP0370090B1 (fr) Immunogenes et peptides biologiquement actifs derives de sequences partagees d'antigenes et d'anticorps anti-idiotypiques ou d'anticorps specifiques contre des recepteurs cellulaires des antigenes
Müller et al. Generation and specificity of monoclonal anti-idiotypic antibodies against human HIV-specific antibodies. I. Cross-reacting idiotopes are expressed in subpopulations of HIV-infected individuals.
JP2000514643A (ja) Ｂ型肝炎表面抗原のａ―決定基の抗原エピトープ及びその利用法
EP0110706A2 (fr) Injection d'anti-idiotype pour obtenir une réponse immunitaire aux agents viraux infectieux
Shearer et al. Structural characterization of viral neutralizing monoclonal antibodies to hepatitis B surface antigen
Lamarre et al. Antiidiotypic vaccination against murine coronavirus infection.
Colucci et al. Interactions between hepatitis B virus and polymeric human serum albumin II. Development of syngeneic monoclonal anti‐anti‐idiotypes which mimic hepatitis B surface antigen in the induction of immune responsiveness
CA1341566C (fr) Reponse immune a des tumeurs induite par des anticorps anti-idiotypes
Kennedy et al. Concepts of idiotype-based vaccines for hepatitis B virus and human immunodeficiency virus
RU2014845C1 (ru) Способ получения вакцины против спида
RICH et al. Induction, via immunization with a monoclonal antiidiotypic antibody, of viral envelope specific Ab3 antisera that neutralize retrovirus infectivity

Legal Events

Date Code Title Description
PUAI Public reference made under article 153(3) epc to a published international application that has entered the european phase

Free format text: ORIGINAL CODE: 0009012

17P Request for examination filed

Effective date: 19890817

AK Designated contracting states

Kind code of ref document: A1

Designated state(s): AT BE CH DE FR GB IT LI LU NL SE

RIN1 Information on inventor provided before grant (corrected)

Inventor name: COLUCCI, GIUSEPPE

Inventor name: WAKSAL, SAMUEL, D.

A4 Supplementary search report drawn up and despatched

Effective date: 19900312

17Q First examination report despatched

Effective date: 19911219

STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: THE APPLICATION HAS BEEN REFUSED

18R Application refused

Effective date: 19930501