IMMUNOLOGICAL USE OF MICROWAVES

Abstract: Antigenic materials are treated with microwaves to destroy immunodominant epitopes and render other, microwave-resistant epitopes more accessible to the immune system. The treated materials (or synthetic microwave-resistant epitopes) may be used as diagnostic or therapeutic agents, or to raise antibodies which in turn may be used as diagnostic or therapeutic agents. The technology is of particular value in the diagnosis and treatment of AIDS.

Description: ΪMMUNOLOGICAL USE OF MICROWAVES Field of the Invention The present invention relates to antigens having microwave-resistant epitopes, and the role of such antigens, and of antibodies to such epitopes, in the diagnosis or treatment of a disease or condition of the body. Background of the Invention Generally, a successful infectious agent is one which modulates the host's immune response to its advantage such that an immunocompetent host responds with an immune response that is not harmful to the infectious agent. Thus, in successful infectious agents, the antigens which cause the predominant immune response, particularly the antibody response, are antigens that are not involved in key metabolic or biological processes fundamental to the survival of the infectious agent in the host. Generally, the antigens which cause the predominant immune response camouflage key infectious agent antigens thus protecting them from the host's immune response. Alternatively, the infectious agent may present antigens which the host recognizes as "self" and consequently does not respond to; such antigens camouflage epitopes borne by antigens essential to the survival of the infectious agent. The HIV-1 virus is known to have antigenie sites on its surface and to leave viral antigens on infected cell surfaces, which mirror normal cellular receptor molecules (e.g., the glycoprotein gpl20, binds to the normal CD4 receptor ) . These epitopes are exposed to the infected host's immune system but are apparently seen as "self" and an antibody response is not usually generated. In order to mount* an effective response against an agent which acts in this way it is necessary to be able to present to the host's immune system key antigens or epitopes thereof against which the host does not normally mount an effective immune response. Current procedures for the production of antibodies to, for instance, HIV include immunization of a host with viral lysates, immunization of a host with reco binant viral proteins, and immunization of a host with peptide antigens (Wang et al . U.S. 4,735,896 and Berzofsky et al. PCT/US87/03471). These procedures for production of antibodies against the human immunodeficiency virus produce antibodies which may neutralize virus in vitro but are not efficacious in vivo. Recent work by Jackson et al . (1988) CThe Lancet, September 17, 647-6513 shows that loss of titre of anti-P24 antibodies correlated with onset of AIDS/ARC in AIDS patients. Passive administration of anti-P24 antibodies provided temporary remission in patients showing loss of anti-P24 titre. Also, HIV is mutating quickly and many variants can be seen even in the same patient during the course of the disease. CSaag et al .. Nature (1988), J4440; Fischer et al.. Nature ( 1988 ) , _____ 4441. It is therefore important to uncover sites on the virus which are essential to the viral lifecycle and consequently less likely to mutate. Knowles, U.S. 4,658,022 (Molecular Diagnostics) teaches denaturing a protein by chemical or physical means to destroy conformational epitopes and expose or enhance the exposure of l near epitopes. Antibodies are prepared against these treatment-res stant epitopes. The methods of denaturation include "heat, soni ation, high or low pH" or treatment with a "chaotropic reagent." ( Col. 8, lines 8-13). There is no reference to denaturation by radiation. Similarly, Milich, U.S. 4,683,136 teaches use of a denaturing agent to destroy a conformational epitope, specifically, of a Hepatitis B virus antigen. The modified antigen is then used on an assay for sample antibodies which react with a conformation-independent determinant. Microwaves have been reported to affect the immune response in vivo. Thus Rao, et al . , Bioelectromagnetics, 6:41-52 (1985) reports that microwaves activated peritoneal macrophages and augmented the IgM antibody response of hamsters to SRBCs. While Rao attributes this phenomenon to a thermal effect of the microwaves, Zafra, et al . , Int. Arch. Allergy Appl . Immunol., 85:478-82 (1988) says that MW activation of macrophages is independent of the heating effect. Other reports belittle the importance of microwaves. Gildersleeve et al . , Comp. Biochem. Biophysiol., 87A:375-80 (1987) reports that the humoral immune potential of Japanese quail is not affected by embryonic exposure to microwaves. Liddle, et al . , Bioelectromagnetics, 7:91-94 (1986) found that low intensity (IMW/c 2 ) microwaves did not alter the circulating antibody response of mice exposed to S. pneumoniae III. Vaccine use of microwaved antigens is known. Injection of mice with microwaved B16 melanoma cell membranes, prior to challenge with B16 melanoma, increased the subjects' survival time as compared with mice immunized with untreated cell membranes. Douss, et al . , C.R. SOC. Biol., 179:615-24 (1985). Gray, U.S. 3,660,234 attenuated a virus by microwave treatment. Microwave irradiation can have a heating effect. Heat has been used to selectively denature epitopes. Hansen, W088/09934 describes a CEA immunoassay in which the sample is heated to 90°C and then assayed using monoclonal antibodies which bind to heat-stable CEA epitopes. Anacker, et al., Infection and Immunity, 55:825 (1987) reports that Rickettsia rickettsi surface proteins exhibit both heat-sensitive and heat-resistant epitopes. Monoclonal antibodies specific for one type or the other have been developed. Only antibodies against heat-sensitive epitopes had a protective effect on immunized mice. "Conceivably, the heat-resistant epitopes are exposed by denaturing influences, such as heat, and are not immunoaccessible to any appreciable extent on the surfaces of normal infectious rickettsiae." (p. 827). See also, Leye and Walker, Infection and Immunity, 56:2587-93 (1988); Sakomoto, et al., 761 Bakt. Hyg. A., 259: 507-19 (1985). However, microwave treatment does not need to heat the subject. See Chen, U.S. 4,327,180. Microwaves have been used to fix cells without antigen loss. See Leong and Milios, J. Pathol . , 148:183-7 (1986); Patterson, Jr. and Bulard, Stain Technology, 55:71 (1980); and Hopgood, et al . , Histoche ical J., 16:1171-91 (1984). No admission is made that any reference is prior art or is material. Descriptions are based solely on the publications and are not binding on applicants. All references in this specification are incorporated by reference to the extent pertinent. Summary of the Invention It is the object of this invention to overcome the deficiencies of the prior art. More particularly, the invention relates to the use of microwave radiation in the preparation of antibodies against immunorecessive (" asied") epitopes of an antigen associated with an infectious agent, and especially against the epitope of an antigen essential to the survival or pathogenicity of the agent. Such antibodies may be used to detect the circulating antigen having such epitopes and thereby detect the presence of the infectious agent. The agent is more reliably detected in this manner, since the normally immunorecessive epitopes are less likely to be altered by strain-to-strain variations or blocked by the presence of circulating antibodies generated against the immunodominant epitopes. The antibodies may also be used in immunotherapy. The microwaved antigens, or the microwave-resistant epitopes thereof, are prepared and used to detect whether the body has mustered an immune response against such epitopes, to raise antibodies against such antigens and epitopes for diagnostic or therapeutic use, and for direct immunization of patients to stimulate an immune response. More specifically, according to this invention there is provided a process for altering the immunological characteristic of an antigenie entity which comprises irradiating an antigenic entity having at least one microwave-resistant epitope and at least one microwave-labile epitope with microwave radiation of an intensity, wavelength and duration effective to reduce the antigenicity of said microwave-l bile epitope while retaining said microwave-resistant epitope, thereby yielding an altered antigenic entity. According to a second embodiment of this invention there is provided a process for altering the immunological characteristic of an antigenic entity which comprises irradiating an antigenic entity having at least one microwave-resistant epitope with microwave radiation of an intensity, wavelength and duration effective to produce a altered antigenic entity retaining said microwave-resistant epitope and providing increased accessibility to said epitope by antibodies which recognize said epitope. According to a third embodiment of this invention there is provided an antibody specific for a natively immunorecessive, microwave-resistant epitope of a naturally occurring antigen. According to a fourth embodiment of this invention there is provided a process for preparing an antibody specific for a microwave-resistant epitope which comprises immunizing an animal with a microwave-heated antigenic entity, said entity natively having at least one microwave-resistant epitope, the antigenicity of said microwave treated antigenic entity differing from that of said entity prior to microwave treatment. According to this invention there is also provided antibodies prepared by the process of the fourth embodiment. According to a fifth embodiment of this invention there is provided a non-naturally occurring antigen characterized by the presence of a microwave resistant epitope common to a naturally occurring antigen and the absence of a microwave-labile epitope common to the naturally- occurring antigen, said naturally occurring antigen being associated with an infectious agent. According to a sixth embodiment of this invention there is provided in an immunoassay for the detection in a sample of an antigen associated with an infectious agent, wherein an antibody reagent which recognizes said antigen is incubated with the sample or a portion thereof, the improvement consisting of providing an antibody reagent specific for a microwave-resistant epitope of such antigen and of subjecting the sample of a portion thereof to microwave-treatment prior to incubating it with said antibody. According to a seventh embodiment of this invention there is provided in an immunoassay for the detection in a sample of an antibody produced in a subject in response to an antigen associated with an infectious agent, wherein an antigenic reagent recognized by said antibody is incubated with the sample or a portion thereof, the improvement consisting or providing a first antigenic reagent which presents essentially only mi rowave-resistant epitopes of said antigen. According to an eighth embodiment there is provided a therapeutic or prophylactic composition comprising an antigen according to the first, second or fifth embodiments or an an antibody according to the third embodiment and a pharmaceutically acceptable carrier. According to a ninth embodiment of this invention there is provided a method of inducing immunity in a host to a condition associated with an antigenic entity comprising administering to said host an immunity inducing amount of a composition according to the eighth embodiment. According to a tenth embodiment of this invention there is provided a method for the in vitro detection and diagnosis of an antigenic entity having at least one microwave-resistant epitope in an animal or human suspected of having the antigenic entity, which method comprises: contacting a sample from said animal or human with antibodies raised against a altered antigenic entity prepared by irradiating the antigenic entity with microwave radiation of an intensity, wavelength and duration effective to produce the altered antigenic entity which retains said microwave-resistant epitope and provides increased accessibility to said epitope by antibodies which recognize said epitope; and determining whether said antibodies have bound to the antigenic entity. According to an eleventh embodiment of this invention there is provided a method for the in vitro detection and diagnosis of an antigeni entity having at least one microwave-resistant epitope and at least one microwave-labile epitope in an animal or human suspected of having the antigenic entity, which method comprises: contacting a sample from said animal or human with antibodies raised against a altered antigenic entity having at least one microwave-resistant epitope prepared by irradiating an antigenic entity having at least one microwave-resistant epitope and at least one microwave-labile epitope antigenic entity, said altered antigenic entity being characterized by the retention of said microwave-resistant epitope and the reduction in the antigenicity of said microwave-labile epitope; and determining whether said antibodies have bound to the antigenic entity. According to a twelfth embodiment of this invention there is provided a method for the in vitro detection and diagnosis of an antigenic entity having at least one microwave-resistant epitope in an animal or human suspected of having the antigenic entity, which method comprises: contacting a sample from said animal or human with antibodies raised against a non-naturally occurring antigen characterized by the presence of a microwave-resistant epitope common to the antigenic entity; and determining whether said antibodies have bound to the antigenic entity. According to a thirteenth embodiment of this invention there is provided an anti-idiotypic antibody specific for an antibody according to the invention. According to a fourteenth embodiment of this invention there is provided a diagnostic kit for the detection of an antigenic entity having a microwave radiation resistant epitope or antibodies to the antigenic entity, comprising: an altered antigenic entity of the invention; together with antibodies of the invention specific for the antigenic entity. According to a fifteenth embodiment of this invention there is provided a diagnostic kit for the detection of an antigenic entity having a microwave radiation resistant epitope, comprising: antiidiotypic antibodies of the invention; together with antibodies of the Invention specific for the antigenic entity. Typically the diagnostic kits of the invention also include: an antigenic entity having a microwave radiation resistant epitope. Monoclonal antibodies of the invention are typically produced by bybridomas formed by fusion of cells from a mouse myeloma line and spleen cells from a mouse prior immunized with an altered antigenic entity of the invention. More specifically, mice are immunized with an altered antigenic entity of the invention and monoclonal antibodies are produced according to the method of Bundesen et al . , 1984, Vet. Immunol, and Immunopath. the contents of which are incorporated herein by cross-reference. Anti-idiotypic monoclonal antibodies of the invention are typically prepared from a monoclonal antibody of the invention (prepared from a hybridoma cell line) by: purifying the monoclonal antibody; injecting the purified monoclonal antibody and acceptable carrier, diluent and/or adjuvant into a suitable host; and preparing anti-idiotypic monoclonal antibodies according to the above described procedure for preparing monoclonal antibodies. The invention provides the following advantages: 1. Identification of epitopes of infectious agents which the infectious agent obscures from the immune system of the host thereby rendering the host incapable of neutralizing or killing the agent by the immune response. 2. Antibodies can be raised in either the natural host or an alternative animal or tissue culture system which are important to the life cycle of the infectious agent. 3. Vaccination against infectious agents against which vaccines prepared by traditional techniques have been ineffective. 4. The ability to immunize passively against an infectious agent and ameliorate symptoms of the disease caused by the infectious agent. In the case of AIDS it is advantageous to return a patient to the remission state whereby the patient achieves im unocompetence and concomitant ability to respond to an active vaccination program. 5. The provision of diagnostic kits which differentiate between those patients with the disease and those who have been immunized against the disease. This is particularly advantageous in the blood banking industry. Brief Description of Drawing Fig. 1 is a flow diagram of the particular combination of steps used in processes of the invention for the development of the antibodies and subsequent vaccines and kits. Detailed Description of the Invention An antigen is any molecule that can generate or be induced to generate an immune response in an appropriate animal. It may be a polypeptide, protein, glycoprotein or a carbohydrate, but is not necessarily restricted to these classes of molecules. An epitope (or antigenic determinant) is the structural feature of an antigen which is recognized fay an antibody. This feature, in the case of a polypeptide, may be formed by a continuous sequence of amino acids, in which case the epitope is said to be linear or sequential, or by the three-dimensional folding of the polypeptide so that portions distant in sequence are brought into close proximity, whereupon the determinant is said to be conformational . Antigens frequently have more than one epitope; certain epitopes elicit a strong immune response and are said to be "immunodomi ant"; others are less significant and are called "immunorecessive". A structure which has the potential to act as an epitope, but 1s not available for antibody recognition until a masking structure is removed, is said to be "cryptic". A "cryptic" epitope may be considered an extreme case of immunorecessivity. The present invention is generally directed to the use of microwave radiation, with or without a heating effect, to render an immunorecessive epitope of an antigen more immunologically accessible. Non-essential epitopes are believed to be more likely to be immunodominant and microwave labile. Epitopes essential to the survival of the infectious agent are more likely to be immunorecessive and microwave resistant. Our reasoning is as follows: (i) An infectious organism cannot escape immune surveillance and therefore it will, by a process of natural selection, present antigenic epitopes that are highly antigenic and can initiate a strong immune response, which is either ineffective or incapable of eliminating the infectious organism before it can be reproductively successful. (ii) Such epitopes can probably mutate quite rapidly and consequently " heir tertiary structure at the molecular level is controlled by weak bonding forces allowing substantial tertiary alteration for the minimum structural alteration. Thus these epitopes are MW liable. (iii ) Molecules or epitopes essential for survival will be hidden from immune surveillance as a process of natural selection. Some of these epitopes will be accessible to the immune system and while they are immunogenic under the appropriate circumstances, they are generally not "seen" or processed by the immune system and thus can be regarded as being immunorecessive. ( iv ) These essential and accessible immunorecessive molecules or epitopes must be highly conserved to prevent changes and it is logical to propose that their tertiary structure would be controlled by strong bonding forces. Thus these essential molecules or epitopes would be more likely to be MW resistant. The body defends itself against infectious agents by immune recognition of antigens associated with those agents. Some examples of intracellular and extracellular infectious agents are provided in Table 1. However, the method can be applied, to production of antibodies against virtually any antigen. Table 2 provides examples of antigens associated with some of the agents of Table 1. For effective passive therapy, the monoclonal antibody is preferably but not essentially of the same species as the recipient when the recipient is immunocompetent (e.g., in the case of a human subject, the monoclonal antibody is preferably humanized or of human origin). The recipient of an antigen or antibody according to this invention may be a human, a nonhuman mammal, a bird, a fish, a reptile or an amphibian, i.e., any vertebrate. Typically, the recipient is a human or a farm animal (e.g., cow, pig, horse, goat, sheep), pet (e.g., cat, dog) or research (e.g., mouse, rat, guinea pig, ape, chimpanzee, rabbit) animal. The antigens to be irradiated may be obtained from nature or prepared synthetically. Thus, a protein antigen may be isolated from its natural source, synthesized directly by joining amino acids, or produced by transforming a host cell with a recombinant DNA molecule which bears an expressible gene encoding the protein, or a molecule cleavable to obtain that protein. Once the microwave-resistant epitopes of an antigen have been identified, it is also possible to dispense with isolating and irradiating the whole antigen, and instead prepare smaller molecules bearing the desired microwave-resistant epitopes. If these molecules are too small to induce an immune response, they may be conjugated to an immunogenic carrier. In the case of a peptide antigen, the conjugate may be expressed by recombinant DNA techniques as a fusion of the desired hapten and carrier protein. The antigens, whether natural or synthetic, may be subjected to other physical or chemical treatments, before or after exposure to microwaves, which affect their immunological characteristics. For example, they may be cleaved with a p oteolytic agent, or partially denatured by heat, sonication, high or low pH, chaotropic agents, non-MW radiation, and the like. The antigens, when irradiated, may still be associated with the infectious agent or infected cell, or they may be in an isolated form. For the purposes of this invention, microwaves are defined as electromagnetic waves having a wavelength primarily in the range of 0.1 to 30 cm (and thus frequencies of 1 to 300 GHz). Typically, the antigens are irradiated for 30 to 180 seconds with intensities of 400 to 700 watts at a frequency of 2.450 GHz. Generally, the total flux of microwave irradiation, of the infectious agent or antigen is not of such a magnitude as to cause proteins therein to denature. Microwave radiation may be produced by backward wave oscillators, klystrons or other suitable devices. Microwave treatment heats its subject by transferring energy to water, which in turn can transfer energy to adjacent antigen molecules; such heating will affect some epitopes. When a heating effect is not desired, the subject is microwaved in the presence of a heat sink such as a volume of water contained in a suitable vessel such as a glass beaker. Typically 200 to 250 L of water in a 250 mL beaker is appropriate. Under these conditions, energy transfer can occur by other processes such as molecular collisions between water molecules and adjacent antigen molecules to cause antigen modification. For the purpose of this invention, the term "microwave-resistant antigen" refers to (1) natural antigens containing only microwave-resistant epitopes, (2) antigens initially having both microwave-resistant and microwave-labile epitopes which have been irradiated to degrade the latter, and (3) synthetic antigens constructed so as to present only the microwave-resistant epitopes of a corresponding natural antigen. It should be recognized that the terms "microwave-resistant" and "microwave-l bile" should not be taken in an absolute sense, e.g., depending on the duration and intensity of the MW exposure, the percentage of molecules in which a microwave-labile epitope is destroyed will vary. Similarly, an antibody may be said to be "specific" for a MW-resistant epitope even though it has some cross-reactivity with other epitopes. For immunological techniques not fully described herein, see Harlow and Lane, ANTIBODIES: A LABORATORY MANUAL (Cold Spring Harbour: 1988). Polyclonal antibodies to microwave resistant epitopes present on the surface of the infectious agent or on the surface of a cell infected with the infectious agent can be prepared by immunizing an appropriate animal using standard immunization procedures and collecting serum. Human polyclonal antibodies may be recovered from the sera of infected individuals having a high titer of the antibody of interest. Antibodies which recognize irradiated and non-irradiated antigen can be purified from the serum by conventional affinity purification techniques. Monoclonal antibodies to microwave resistant epitopes present on the surface of the infectious agent or on the surface of a cell infected with the infectious agent can be prepared by immunizing mice or rats or human lymphoid cells in tissue culture with microwave irradiated antigen. In the murine system this would be performed by obtaining spleen cells from the immunized animal. The spleen cells are then fused with myeloma cells to produce hybridomas. The hybridomas are screened and cloned to select those hybridomas producing antibodies which recognize irradiated and non-irradiated antigen. Screening of the hybridomas is performed using .both irradiated and non-irradiated antigen preferably. In one embodiment, these antibodies are used, for passive immune treatment of a host. Passive immunotherapy for AIDS patients with polyclonal or monoclonal antibodies is described by arpas, U.S. 4,863,730. Monoclonal antibodies, because of their specificity are preferred. These monoclonal antibodies may be able to mediate natural host immune responses, such as antibody dependent cellular cytotoxicity (ADCC) or complement mediated lysis. Alternatively, they may be conjugated to a che otherapeutic agent or toxin, such as ricin. This invention provides a process for preparing antibodies against an immunorecessive epitope of an infectious agent or antigen, comprising: (a) treating the infectious agent or antigen with microwave radiation to render immunorecessive epitopes on the infectious agent or antigen immunologically accessible; (b) raising antibodies against the now more immunologically accessible epitopes on the infectious agent or antigen; and (c) screening the raised antibodies to select specific antibodies to microwave radiation resistant epitopes present on native antigen and the treated antigen. The process includes screening the specific antibodies to select antibodies to microwave radiation resistant epitopes present on the surface of the infectious agent or on the surface of a cell infected with the infectious agent and accessible to a humoral, cell-mediated or a combination of humoral and cell-mediated immune response in a host. The antibody may be associated with a therapeutic agent or chemotherapeutic agent (such as ricin) effective against the infectious agent or infected cells and used to direct the therapeutic or chemotherapeutic agent to kill or immobilise the infectious agent or infected cells. The antibody may be a human, murine, bovine, porcine, rabbit, goat or other species of monoclonal antibody, or a chimeric antibody in which the constant and variable regious are derived from different species (e.g., human constant and murine variable). In another embodiment, the antibodies are used diagnostically for the detection of antigens which bear microwave-resistant epitope. In this embodiment, a sample believed to contain the antigens of interest is subjected to microwave treatment, with or without preliminary separation or modification steps, and assayed using the antibodies as reagents. Any assay format conventional in the immunological arts may be used, including both sandwich, competitive assays. The antibody may be insolubilized directly or indirectly, covalently or noncovalently, on an insoluble support, or it may be labeled with a convenient label, such as a radioisotope, fluorophore or enzyme. It may also be immobilized on an agglutinable particle of a kind conventionally used in agglutination assays, such as a latex bead. The samples to be assayed using the reagents of this invention may be biological samples, such as tissues and biological fluids (e.g., blood, urine, milk, etc. ) or nonbiological samples such as soil, water, or food. It is contemplated that specifically binding fragments of antibodies may be substituted for whole antibodies in the present invention. The antibodies are also useful in identifying those microwave resistant epitopes that may be present in or on the surface of both the infectious organism and normal cells of the infected host. A host immune response to such epitopes can lead to autoimmune disease processes. The antibodies are also useful in the purification of the microwave-resistant antigens and the characterization of their microwave-resistant epitopes. For purification, the antibodies are associated with an affinity chromatography support. The epitopes may be localized by preparing fragments of the antigen (either by proteolysis of the whole antigen or by direct production of the fragment) and testing different fragments for affinity to the antibodies of this invention. The amino acid sequence data may also be used to predict antigenic sites for further analysis. As a result of the characterization of the epitopes, synthetic analogues may be designed which present the desired microwave resistant, natively immunorecessive epitopes in a more immunologically accessible form than in__the native antigen. The antigens of this invention have a variety of uses. First, they may be used in labeled or insolubilized form in a diagnostic kit for the detection of circulating antibodies reacting with microwave resistant epitopes present on both native and treated antigen which is not seen by the normal immune response in a host which allows detection of whether the host has been vaccinated against an infectious agent. Such a kit comprises antigens having microwave resistant epitopes and the corresponding monoclonal antibodies or affinity purified polyclonal antibodies as positive control. The kit may additionally include native antigens to permit detection of the normal antibody response and thereby allow discrimination between infected hosts and vaccinated hosts. Second, the antigens may be used therapeutical ly. By rendering an epitope immunologically accessible and raising monoclonal antibodies against such epitopes it is possible to identify candidate epitopes for the provision of recombinant vaccines comprising the epitope(s) that have been demonstrated to be unique to the infectious organism and able to mediate a protective immune response, while omitting those epitope(s) that have been demonstrated to be normal host cell autoantigen(s) that have the potential to produce autoimmune disease. The epitope may also be provided in the form of the infectious agent or antigen itself after treatment to render the relevant epitope immunologically accessible or may be provided in combination with other epitopes of the native infectious agent or antigen, to provide a vaccine. In this example, care is preferably taken to eliminate microwave resistant epitopes that are common to both the infectious organism and the host, in order to preclude the possibility of generating an autoimmune response when using such a vaccine. Anti-idiotypic antibodies may be used for the same purposes as antigens. Anti-idiotypic antibodies which mimic the microwave resistant epitopes present on the surface of the infectious agent or on the surface of a cell infected with the infectious agent can be prepared by immunizing an immunocompetent host with said monoclonal antibodies; harvesting antibodies against said monoclonal antibodies from said host; and screening the harvested antibodies to detect those antibodies which are anti-idiotypic antibodies. Thus, the vaccines of the present invention may be (a) a microwave-treated infection-related natural antigen that has been treated to remove epitopes common to both the infectious organism and the host, (b) a synthetic or recombinant antigen presenting a microwave resistant epitope of a unique natural infection-related antigen that is not a shared autoantigen of the host, or (c) an anti-idiotypic antibody raised against an antibody specific for a unique microwave-resistant epitope of an infection-related antigen that is not a shared autoantigen of the host. Preferably, the antigen lacks one or more masking epitopes of an analogous antigen of the infectious agent of interest, so that the immune response to other epitopes of the antigen is enhanced. A plurality of antigens presenting the same or different microwave-resistant epitopes, may be administered to achieve the desired immune response. The immunization agent may include an adjuvant such as aluminum hydroxide. Microwave-resistant, infection-specific antigens may be of particular value in the treatment of viral infections associated with immunoparalysis and/or autoimmune reactions. HIV-1 is now generally accepted as being the etiologic agent in the development of acquired immune deficiency syndrome (AIDS). It is suspected that other disease states displaying autoimmune phenomena where there is an involvement of anti-lymphocyte antibodies (Juvenile Rheumatoid Arthritis, JRA; Systemic Lupus Erythematosus, SLE), or auto-antibodies to other cell types (Type 1 Diabetes Mellitus; Multiple Sclerosis, MS; Graves Disease; Myasthenia Gravis) may also have a viral aetology. Sera from HIV-1 infected individuals has been shown to contain antibodies able to mediate antibody-dependent cellular cytotoxicity (ADCC) reactions on in vitro HIV-1 infected cells. In general, serum ADCC activity decreased as the disease progressed from healthy HIV-1 positive patients to late stage AIDS patients with opportunistic infections and the magnitude of the ADCC response appeared to correlate with the presence of antibody to the gag antigen p24 by immunoblotting. (Rook, et al . 1987). Viral infections are a known cause of transient iymphocytotoxic antibodies which can result in T lymphocytopenia, which usually resolves during recovery from the infection (De Horatius). Similarly, in HIV-1 infected individuals, auto-antibodies to lymphocytes as well as other cell types have also been reported (Kloster, et al . 1984; Dorsett, et al . , 1985; Kiprov, et al . 1985; Williams, et al . 1984; Tomar, et al . 1985). Cytotoxic complement fixing IgG anti-lymphocyte antibodies are present in approximately 90% of AIDS patients. Up to 50% of ARC patients have also been found to have anti-lymphocyte antibodies. These anti-lymphocyte antibodies have been variously identified as specific for the helper/ inducer 0KT4, OKT 11 positive T cells, or reactive with all T cell sub-sets and B cells. Some 18% of AIDS patients have also been found to have autoantibodies that react with red blood cells (Toy, et al . 1985) and autoantibody mediated thrombocytopenia has been found to be a major hematological abnormality associated with AIDS (Strieker, et al . 1985). Virus specific cytotoxic T lymphocytes, capable of killing infected cells, have been widely reported and are believed to be an important mechanism for host defense against infection. HIV-1 specific cytotoxic T cells have, also been found in infected individuals that are able to preferentially kill either autologous B cells expressing recombinant HIV-1 vaccinia virus antigens or autologous HIV-1 infected macrophages and thus indicate that the MHC may be important in restricting the cytotoxic response. Although the major response appears to be against HIV-1 env products, there is also a demonstrable but lesser gag specific cytolytic response (Walker, et al . , 1987). Recently, HLA class-I-restricted gag-specific cytotoxic lymphocytes were found in 13 out of 15 healthy HIV-1 seropositive patients using a recombinant vaccinia virus to express the gag protein of HIV-1. Further, cytotoxic lymphocytes from one patient recognized a 15 amino acid peptide in ρ24 in association with HLA-B27. The finding of gag specific cytolytic T cells in HIV-1 infections is not surprising as a number of other viral infections (e.g., influenza) have also been shown to elicit cytotoxic T cell responses to internal viral structural proteins and the gag gene encoded target proteins were demonstrated to be on the infected cell surface. However, with HIV-1 infection, it is not clear whether the observed gag specific cytotoxic T cell responses exert a beneficial role for the duration of the disease; they may in fact perform a harmful role by mediating autoimmune cytotoxicity. The presence of both antibodies able to mediate ADCC reactions on HIV-1 infected cells that appear to correlate with the presence of gag specific antibodies and cytotoxic T cells that respond to gag antigens coupled with the fact that gag antigens have been found on the infected cell surface in other viral systems, would seem to imply that HIV-1 gag antigens must also be found on the surface of HIV-1 infected cells and that the immune system is able to respond to these antigens. By modifying HIV-1 antigens, we have generated a number of murine MAb that recognize the gag p24 antigen that fall into two distinct types. Type 1 MAb react with crossreacting p24 epitopes that are present on the surface of both HIV-1 infected and uninfected peripheral blood lymphocytes, and red blood cells. One of these Type 1 monoclonal antibodies, H69.67.2D4, also enhanced in vitro HIV-1 plaque formation. Type 2 MAb react with p24 epitopes that are only expressed on HIV-1 infected lymphocytes and are absent from normal uninfected peripheral blood lymphocytes or other normal cells. Some MAb or both Type 1 and Type 2 are able to mediate in vitro ADCC reactions on HIV-1 infected lymphocytes. It is known that there is an antibody spectrum shift with time in animals used for the production of specific antisera that receive repeated booster immunizations. Further, we (Bundesen, et al . 1984) and other groups (Morgar, AU-A-74496/87) routinely down-regulate the normal immunodominant epitope responses to a particular antigen by immunizing mice with antigen absorbed with MAb generated from a prior fusion using native ( unabsorbed) antigen. In this way, the normal immunodominant epitopes are "masked", allowing the production of novel MAb that react with normally immunorecessive or silent epitopes on these antigens. Arguing by analogy, with persistent viral infections such as HIV-1, there is continual antigen presentation and processing of antigen/antibody complexes by the immune system. Over a period of time, formerly immunorecessive epitopes are finally able to generate an immune response and this mechanism would particularly facilitate the production of antibodies to crossreacting virus/self autoantigens. Monoclonal antibodies have been generated that bind to a phosphoprotein of measles virus or a protein of herpes simplex virus type 1 and crossreact with an intracellular intermediate filament protein of human cells, which was thought to be vimentin. This observation gave rise to the concept of "molecular mimicry" and the proposal that viruses could induce autoimmune responses through shared determinants normally present on host cells (Fujinami, et al . 1983). Indirect evidence for this concept is perhaps found in SLE, where monoclonal autoantibodies have been derived from both murine and human systems. Such monoclonal anti-DNA autoantibodies show remarkable poly-specificity; not only do they react with DNA, but with phospholipids such as cardiolipin and proteins such as vimentin. Some 66% of SLE patients have anti-cardiolipin antibodies. Autoantibodies directed against normal pancreatic islet cells has been found in Juvenile diabetes as a result of Coxsackie virus infection and the B cell surface expression of retrovirai protein "neoantigens" has been proposed as a mechanism for stimulating their autoimmune elimination in non-obese diabetic mice. The finding of shared epitopes on normal uninfected peripheral blood lymphocytes, red blood cells and the HIV-1 gag p24 antigen, was crucial to the formulation of our theory. This theory postulates an antibody masking effect of HIV-1 antigens, leading to the down regulation of existing antibody producing clones. When this process is coupled with a time dependent antibody spectrum shift, it leads to the generation of autoantibodies able to bind the crossreacting viral gag/self cell surface epitopes. The autoantibodies generated by this mechanism, as well as mediating direct autoimmune phenomena (ADCC, C mediated lysis) on various target cells such as lymphocytes, red blood cells, platelets, and peripheral nerve cells could also, with further immune processing of cell bound antibody or antibody/antigen complexes, generate cytotoxic T cells reactive to these viral/self epitopes expressed on normal uninfected cell populations. Further, some of these antibodies could be enhancing and thus provide an alternative mechanism for viral infection in non-CD4 bearing cell types. Thus this overall process would provide a simple unifying mechanism to explain the totality of HIV-1 pathogenesis and should allow rational and effective strategies for immune modulation of the disease. Importantly the identification of unique HIV-1 gag epitopes expressed only on the surface of infected cells, could pave the way for the construction of safe and effective vaccines that would only contain the unique epitopes and eliminate crossreactive epitopes. Further, the Type 2 murine monoclonal antibodies that react with the unique gag epitopes may provide an effective means of passive immunotherapy in AIDS patients. Other Type II monoclonal antibodies to microwave resistant epitopes may also be present in stable regions of envelope antigens, such as the region of gp41 with the amino acid sequence RILAVERYLKDQQLLGIWQCSGK, as demonstrated by the Mab H69.67.2A2.26. While this theory was initially proposed to explain AIDS pathogenesis, it has obvious wider relevance to other autoimmune diseases such as MS and SLE. Recently, gag and env HTLV-1 related sequences were detected in peripheral blood mononuclear cells from MS patients and antibodies that react with the gag p24 HTLV-1 antigen have also been found in the serum and cerebrospinal fluid. Consequently, it is highly likely that crossreacting p24 epitopes will be found on the normal cell surface of oligodendrocytes and that such epitopes would mediate an autoimmune response. EXAMPLE 1 Development of DiaQnostic Kit for Circulating Di ofilaria Immitis (Dog Heartworm) Anticien As described above, antibodies developed against epitopes of microwaved antigens that are present on both the untreated and microwaved antigen may be used for the development of diagnostic kits for applications such as the detection of circulating heartworm antigen in dog plasma without the need to process the circulating antigen in order to remove or destroy the autologous antibody that would block test antibody from binding. The following example illustrates this feature. D. immitis Antigen Preparation Adult P. immitis worms were removed from the right ventricles and pulmonary arteries of infected dogs using sterile procedures. The worms were extensively washed in sterile saline and 150 worms were cultured in 300 ml RPMI 1640 medium supplemented with 100 IU/ml penicillin and 100 μg/ml streptomycin for 48 hours with one change of medium after 24 hrs, in a humidified, 5% C0 2> 95% air atmosphere at 37 = C. After 48 hours of incubation, the worms were removed from the culture medium and homogenized on ice for 2.5 min in a total volume of 50 ml PBS. The homogenized P. immitis material was then centrifuged for 15 min at 1,000 g at room temperature to pellet the particulate material and the supernatant containing soluble P. immitis antigens was carefully removed and stored. Immunization Female BALB/c mice were immunized with 2 subcutaneous injections of 10 μg (protein) microwaved D. immitis antigen in Freunds complete adjuvant at 2 weekly intervals followed by a final intraperitoneal injection of 10 μg antigen in 0.5 ml PBS 4 days before the mice were killed and their spleens removed for fusion. Hvbridoma Production The spleens were removed aseptically from 3 immunized mice and the pooled spleen cells were fused with NS-1 myeloma cells (Bundesen et al . , 1984, Vet. Immunol, and Immunopath.). Selected crude clones chosen on the basis of the screening assay were subcloned by limiting dilution and subsequently a number of MAb secreting hybridoma cell lines were finally chosen which recognized both irradiated and non-irradiated D. immitis antigen for expansion to mass culture and for the production of ascitic fluids. The hybridoma cell lines D44.8.3B4/64 and D46.8.4D4/23, which bind to different epitopes on both microwaved and unmicrowaved heartworm antigen were produced and selected in this manner. Ascitic Fluid Production and MAb Conjugation Female BALB/c mice were given an intraperitoneal injection of 0.5 ml 2, 10, 14, tetramethylepentadecane (Pristane, Aldrich Chemical Corp., Milwaukee, Wisconsin) 14 days prior to the injection of 2 x 10 viable hybridoma cells and ascites fluids were collected from the mice 12 to 14 days after injection of the ceils. The ascitic fluid was clarified by centrifugation and MAb were recovered by precipitation with 45% ammonium sulphate and stored at either 4°C or -70°C in phosphate buffered saline (PBS) containing 0.01% sodium azide. Application of Microwaved Antigen Derived MAb to Detect Circulating Parasite Antigen In order to detect circulating antigen derived from an infectious agent in a blood, serum or plasma sample in the presence of an anti-infectious agent host antibody response, test antibodies must either see epitopes not recognized by the host antibody response, or recognize new epitopes in an antigen that has been modified in some manner. Alternatively, the epitope may be stable to the modification process but any host antibody binding to that epitope will be stripped off and denatured, allowing test monoclonal antibody to bind. The process of microwaving antigen allows the production of monoclonal antibodies that can be applied to these types of tests. Enzyme-Immunoassav Serum or plasma samples are diluted 1/5 in phosphate buffered saline (PBS) and microwaved for 90 seconds on the high setting in a domestic microwave oven. The microwaved samples are then reacted with the wells of 96 well microtitre plates coated with the monoclonal antibody, which was given the reference code D46.8.4D4/23, for 60 minutes at room temperature, followed by 3 washes with PBS/Tween (PBST). Horseradish peroxidase conjugated D46.8.4D4/23 was then added and incubated for 60 minutes, followed by a further 3 washes with PBST and ABTS substrate was then added and incubated for a further 20 minutes before stopping the reaction with 0.4% oxalic acid and reading the OD at 420 n on a plate reader. A positive sample from a heartworm infected dog will give an OD greater than the appropriate negative control, whereas a negative sample from an uninfected dog will have an 0D less than or not significantly greater than the negative control Table 3). Latex-Immunoassay Ten μl of untreated serum or plasma are added to 25 μl latex beads coated with the monoclonal antibody D44.8.3B4/64 on a suitable glass or plastic slide, rocked by hand for 3-5 minutes and then assessed for agglutination. A positive sample from an infected dog will agglutinate whereas a negative sample from an uninfected dog will remain milky. EXAMPLE 2 A process for preparing antibodies against immunorecessive epitopes of HIV which are expressed on the surface of HIV and/or expressed on the surface of a host cell infected with HIV such as HIV infected lymphocytes is given below. Appropriate immunizing antigens which can be prepared or isolated by known techniques include: (a) Live HIV infected cells; (b) Live HIV; (c) Selected HIV component; (d) Recombinant HIV component; and (e) Synthetic HIV component. At least one of (a)-(e) is treated with microwave radiation (eg 1ml of a one mg per ml protein solution of (b) + (c) in a Nunc Cryotube microwaved for 60 - 120 seconds at 680 watts at 2450MHz, using a heat sink such as a container of 250ml water to maintain the temperature of the sample below 56 -60°C so as to be below protein denaturing temperature) to render immunorecessive epitopes of the antigen immunologically accessible. Antibodies are raised against the immunologically accessible epitopes on the microwave treated antigens by known techniques (e.g. standard monoclonal antibody production techniques). The antibodies are screened by immunoassay (e.g., enzyme immunoassay or radioimmunoassay) to select specific antibodies to microwave radiation resistant epitopes present on native HIV antigens and microwaved HIV antigens (e.g. gp!20, gp41 and p24. The specific antibodies are further screened to select antibodies to the microwave radiation resistant epitopes which are not recognized by the normal human immune response in HIV infected individuals. The antibodies resulting from this further screening can be used in the construction of immunodiagnostic kits (e.g., enzyme immunoassay or radioimmunoassay or red cell agglutination assay) for the detection of circulating HIV antigens (e.g., gp!20, gp41 and p24) in infected individuals. The antibodies to the microwave radiation resistant epitopes are screened further to select antibodies to unique HIV epitopes present on the surface of a cell infected with HIV and accessible to a humoral, cell-mediated or a combination of humoral and cell-mediated immune response in an infected human and to identify further those monoclonal antibodies that react with both microwaved or unmicrowaved HIV-1 antigen and epitopes present on normal uninfected human cells such as red blood cells, platelets, lymphocytes and other cell types. The unique aπti-HIV-1 antibodies obtained from these latter screenings can be directly administered (e.g., intravenously) to an affected AIDS/ARC patient for passive immune therapy. The antibodies obtained from these latter screenings can further be used to identify the specific epitopes and construct either peptide or recombinant antigens for use as a vaccine. Ideally, these peptide or recombinant antigens would only contain unique HIV epitopes and exclude HIV and normal cell surface crossreacting epitopes, to prevent the possibility of stimulating an autoimmune response as a consequence of immunization. Alternatively, the final vaccine formulation can include a combination of native epitopes and modified epitopes ( e.g., native pi7, p24, gp41 and gp!20 and microwave treated HIV antigen ) . Ideally, such native HIV antigens would also be treated in a manner to eliminate the previously mentioned HIV/normal cell crossreacting epitopes. Alternatively, the antibodies reacting with the unique HIV epitopes can be used to produce antiidiotypic antibodies which can be used as a vaccine. The peptide or recombinant antigens or the antiidiotypic antigens may be used singly, in combination or alternatively in combination with other elements derived from the virus (e.g., microwaved HIV, microwaved HIV components or microwaved recombinant HIV antigens) to provide composite vaccines. The unique antibodies selected in the last screening step or antiidiotypic antibodies can be used to construct immunodiagnostic tests which will discriminate between HIV immunized and HIV infected individuals. Alternatively, the microwave modified antigen could be used to discriminate between HIV infected patients and vaccinated patients. Such tests would have application in blood bank screening. With the exception of antiidiotypic antibodies which are monoclonal any of the above described antibodies could be polyclonal antibodies, produced from suitable experimental animal species or murine or human monoclonal antibodies. HIV Antigen Preparation Several 1 ml aliquots (1 mg/ml in PBS) of human immunodeficiency virus were microwaved for 60 seconds in a standard domestic microwave oven (for example, a Sanyo model EM 3310) with a 250 ml beaker filled with water as a heat sink. This treatment caused both neutralization of viral infectivity and a rearrangement of the viral proteins and other molecules. Alternatively, 300 μl "HTLV-III" viral detergent lysate antigen (800 μg/ml) was microwaved under identical conditions. Immunization Three groups of mice were immunized in the following manner: Group A: Six female BALB/c mice were each immunized twice with subcutaneous injections of 10 μg microwaved HIV-1 virus in 0.05 ml of PBS emulsified with 0.05 ml Freunds complete adjuvant (FCA), at weekly intervals. Four days before the mice were killed and their spleens removed for fusion, a final intraperitoneal injection was given consisting of 10 μg of antigen In 0.5 ml PBS. Group B: Six female BALB/c mice were each immunized twice with subcutaneous injections of a mixture of 5 μg of microwaved HIV-1 virus and 5 μg microwaved "HTLV-III" viral detergent lysate antigen in 0.05 ml PBS emulsified with 0.05 ml FCA, at weekly intervals. Four days before the mice were killed and their spleens removed for fusion, a final intraperitoneal injection was given consisting of 5 μg of each antigen in 0.5 ml PBS. Group C: Six female BALB/c mice were each immunized twice with subcutaneous injections of a mixture of 5 μg of microwaved HIV-1 virus and 5 μg "HTLV-III" viral detergent lysate antigen in 0.05 ml PBS emulsified with 0.05 ml FCA, at weekly intervals. Four days before the mice were killed and their spleens removed for fusion, a final intraperitoneal injection was given consisting of 5 μg of each antigen in 0.5 ml PBS. Hybridoma Production The spleen was removed aseptically from one mouse from each of the 3 immunization groups and pooled. A single cell suspension was prepared and the pooled spleen cells were fused with NS-1 myeloma cells (Bundesen et al . 1984, Vet. Immunol, and Immunopath.). Selected crude clones chosen on the basis of the screening assay were subcloned by limiting dilution and subsequently a number of MAb secreting hybridoma cell lines were finally chosen for expansion to mass culture and for the production of ascitic fluids. All hybridoma supernatants were tested for immunoreactivity to the following antigens using standard enzyme immunoassay procedures: ( i ) microwaved HIV-1 virus grown in CEM cells (ii) inactivated HTLV-III detergent lysate antigen (iii) inactivated HIV-1 detergent lysate antigen (iv) microwaved inactivated HIV-1 detergent lysate antigen (v) uninfected CEM cell lysate Results: Seventy-four primary monoclonal antibody secreting clones were produced, over 50% of which reacted with p24 by western blot analysis. Immunofluorescent studies indicate that these antibodies recognized epitopes on the surface, of HIV-1 infected cells. Of these clones, some 24 were chosen for further analysis (Table 5). Monoclonal antibodies in accordance with the invention can be utilized for recognition of epitopes for production of synthetic or recombinant vaccines. By making monoclonal antibodies to microwaved HIV-1 antigens, a unique panel of monoclonal antibodies were generated that reacted with various p24 and other HIV-1 epitopes, e.g., gp41 , by western blot and by EIA on normal and microwaved HIV-1 antigens and recombinant antigens. Using selected monoclonal antibodies from this panel, p24 epitopes were discovered on the surface of in vitro HIV-1 infected cells that were absent from uninfected cell surfaces. Monoclonal antibodies displaying this type of response were classified as Type 2 antibodies and were able to mediate in vitro antibody dependent cellular cytotoxic (ADCC) reactions on HIV-1 infected cells (Table 6). Other selected monoclonal antibodies from this panel recognized crossreacting p24 epitopes that were present on both HIV-1 infected cells and normal human cell types e.g. red blood cells, platelets, peripheral blood lymphocytes and brain and nerve tissue. Such antibodies were classified as Type 1 antibodies and were able to mediate ADCC reactions on both HIV-1 infected (Table 6) and uninfected cells (Table 7). A method is proposed for the generation of monoclonal antibodies from microwaved viral antigens and/or microwaved viral antigenes that have been absorbed with monoclonal antibodies made previously to either native or microwaved viral antigens. Some monoclonal antibodies generated in this manner will identify viral and human cell surface crossreacting epitopes (Table 1 MAb) and may mediate ADCC and/or C* dependent lytic reactions. Other monoclonal antibodies will identify viral antigens expressed only on the surface of infected cells (Type 2 MAb) and some of these monoclonal antibodies will be able to mediate ADCC and/or C dependent lysis reactions. A further method is proposed for the production of effective anti-viral peptide or recombinant vaccines that contain viral epitopes that are only expressed on the surface of infected cells from which the specific cell surface crossreacting epitopes have been identified and eliminated. Thus a "safe" vaccine may be produced that will protect against disease, rather than enhancing infection or causing autoimmune disease or immune paralysis. A third method is proposed for the immune modulation of the autoimmune disease or immune paralysis by treatment with antiidiotypic antibodies to the specific crossreactive MAb, which could act to switch off the autoimmune antibody response. Similarly, recombinant antigens or peptides identified by the specific crossreactive monoclonal antibodies could also be used to down-regulate the autoimmune response. This approach could be employed for HIV-1 and other retroviruses (including those recently found to be associated with MS, SLE, etc.). Industrial Applicability The methods of the invention are useful in the production of monoclonal antibodies against an antigen or an epitope of an antigen which is immunorecessive. The methods of the invention are also useful in the production of anti-idiotypic antibodies raised against these monoclonal antibodies. The antibodies of the invention can be used in diagnostic kits, therapeutically or for vaccination purposes. Diagnostic kits in accordance with the invention allow differentiation between patients vaccinated against an infectious agent and patients infected with the disease. Monoclonal antibodies in accordance with the invention can be utilised for recognition of epitopes for production of synthetic or recombinant vaccines. Additional References 1. Bundesen, P.G., et al . , Monoclonal antibodies directed against Brucella abortus cell surface antigens. Vet. Immunol. Immunopathol., 1984; 8:245-260. 2. Dorsett B. et al . Anti-lymphocyte antibodies in patients with the acquired immune deficiency syndrome. Amer J Med 1985; 78:621-626. 3. Fujinami, R.S., et al . Molecular mimicry in virus infection: Crossreaction of measles virus phosphoprotein or of herpes simplex virus protein with human intermediate filaments. Proc. Nat. Acad. Sci . USA. 1983; 80:2346-2350. 4. Kiprov DD et al . Antilymphocyte antibody and seropositivity for retroviruses in groups at high risk for AIDS. New Eng J Med, 1985; 312:1517. 5. Kloster BE et al . Lymphocytotoxic antibodies in the acquired immune deficiency syndrome (AIDS). Clin Immunol and Immunopath 1984; 30:330-335. 6. Rook, A.H., et al. Sera from HTLV-III/LAV antibody-positive individuals mediate antibody-dependent cellular cytotoxicity against HTLV-III/LAV-infected T cells. J. Immunol. 1987; 138:1064-1067. 7. Strieker RB et al . Target platelet antigen in homosexual men with immune thrombocytopenia. New Eng J Med, 1985; 313:1375-1380. 8. Tomar, R.H. et al . Cellular targets of antilymphocyte antibodies in AIDS and LAS. Clin. Immunol. Immunopath., 1985; 37:37-47. 9. Toy PTCY et al . Positive direct antiglobulin test associated hyperglobuliπemia in acquired immunodeficiency syndrome (AIDS). Amer J Hematol, 1985; 19:145-150. 10. Walker, B.D., et al . HIV-specific cytotoxic T lymphocytes in seropositive individuals. Nature, 1987; 328:345-348. 11. Williams JR et al . Lymphocyte-reactive antibodies in acquired immune deficiency syndrome. J Clin Immunol, 1984; 4:118-123. Deposits of Hybridomas The following hybridomas were deposited at the European Collection of Animal Cell Cultures'(ECACC), Vaccine Research and Production Laboratory, Public Health Laboratory Service, Centre for Applied Microbiology and Research, Porton Down, Salisbury, Wiltshire, United Kingdom SP4 OJG on 2 February 1990: 1. Murine hybridoma H46.8.4D4/23 Produces monoclonal antibodies with anti-Pi rofil aria immitis specificity. Accession no. 90020301 2. Murine hybridoma H68.67.1B4/2 Produces monoclonal antibodies with anti-HIV 1 p24 antigen specificity. Accession no. 90020302 3. Murine hybridoma H68.67.2B1/36 Produces monoclonal antibodies with anti-HIV 1 p24 antigen specificity. Accession no. 90020303 4. Murine hybridoma H69.67.2A2/26 Produces monoclonal antibodies with anti-HIV 1 gp41 antigen specificity. Accession no. 90020304 5. Murine hybridoma H70.67.3A5/63 Produces monoclonal antibodies with anti-HIV 1 p24 antigen specificity. Accession no. 90020305 SUBSTITUTESHEET TABLE 1 INTRACELLULAR INFECTIOUS AGENTS EXTRACELLULAR INFECTIOUS AGENTS Viruses: Bacteria: e.g. HIV, Rubella, Rotaviruses, e.g. Neisseria gonorrhoeae. Hepatitis, Herpes, Rhinoviruses, Bordetella pertussis τ Ortho- and para-myxoviruses, Vibrio cholerae, Epstein-Barr, Cytomegalovirus, Salmonella typhimurium Picornaviruses, Polyoma, Papilloma and Retro viruses. Spirochetes: Worms: e.g. Syphilis and Leptospirosis e.g. Heartworm, Schistosomes Protozoa: Arthropods: e.g. Malaria e.g. ticks, fleas - 38 - TABLE 2 INTRACELLULAR INFECTIOUS EXTRACELLULAR INFECTIOUS AGENTS AGENTS Vi ruses : Bacteria: Core anti gens Surface antigens e . g. HIV-1 p55 , p40, p24 Flagella antigens pl7 Surface antigens Worms: e.g. HIV-1 gp41 , gp!20 Surface antigens Spirochetes: Gut antigens Nuclear antigens Enzyme antigens Surface antigens Arthropods: Protozoa: Gut antigens Nuclear antigens Enzyme antigens Surface antigens TABLE 3 Dog Heart Worm Enzyme-Immunoassay Results from Necropsied Dogs Test Positive Test Negative 95 8 2 76 Test Sensi tivi ty Test Specifi ci ty Test Accuracy 92.2% 97.4% 94.5% (95/103) (76/78) (171/181) TABLE 4 Dog Heartworm Latex Agglutination Results for Necropsied Dogs Condition of Dogs Number Test Positive Test Negative Infected 110 92 18 Uninfected 53 1 52 Test Sensitivity Test Specificity Test Accuracy 83.6% • 98.1% 88.3% (92/110) (52/53) ( 144/163) TABLE 6 CLONE RBC PLAT LYMP P24 WB ADCC BDG BDG BDG EIA % OD RBC BDG %, percent red blood cell binding PLAT BDG %, percent platelet binding LYMP BDG %, percent peripheral blood lymphocyte binding p24 EIA, optical density of recombinant p24 antigen EIA WB, western blot analysis ADCC %, percent antibody dependent cellular cytotoxicity TABLE 7 CLONE CPM Cr 51 RELEASE HIV-1 INFECTED UNINFECTED H68.67.1 B4/2 4845 4881 H68.67.2D2/ 19-84 5399 5138 H69.67.2D4/126 5263 5065 NEGATIVE CONTROL 2546 2479

Claims: CLAIMS 1. A process for altering the immunological characteristic of an antigenic entity which comprises irradiating an antigenic entity having at least one microwave-resistant epitope and at least one microwave-labile epitope with microwave radiation of an intensity, wavelength and duration effective to reduce the antigenicity of said microwave-labile epitope while retaining said microwave-resistant epitope, thereby yielding an altered antigenic entity. 2. The process of claim 1, wherein the antigenicity of said microwave-labile epitope is destroyed, but said microwave-resistant epitope is still immunologically recognizable. 3. A process for altering the immunological characteristic of an antigenic entity which comprises irradiating an antigenic entity having at least one microwave-resistant epitope with microwave radiation of an intensity, wavelength and duration effective to produce a altered antigenic entity retaining said microwave-resistant epitope and providing increased accessibility to said epitope by antibodies which recognize said epitope. 4. The process of any one of claims 1-3 wherein the antigenic entity is an infectious agent. 5. The process of claim 4 wherein the infectious agent is a virus. 6. The process of claim 1 wherein the microwave-resistant epitope is formed by a first antigen of said entity and the microwave-labile epitope is formed by a second and different antigen of said entity. 7. The process of claim 1 wherein the microwave-resistant and microwave-labile epitopes are formed by the same antigenic molecule, 8. The process of claim 1 wherein the microwave-labile epitope was, prior to the irradiation, an immunodominant epitope of the entity. 9. The process of claim 1, wherein the microwave-resistant epitope was, prior to the irradiation, an immunorecessive epitope of the entity. 10. The process of claim 5 wherein the virus is HIV. 11. The process of claim 4 wherein the infectious agent is a parasite. 12. The process of claim 11 wherein the parasite is heartworm. 13. An antibody specific for a natively immunorecessive, microwave-resistant epitope of a naturally occurring antigen. 14. The antibody of claim 12, further characterized as a monoclonal antibody. 15. A altered antigenic entity obtained by the process of any one of claims 1-12. 16. A process for preparing an antibody specific for a microwave-resistant epitope which comprises immunizing an animal with a microwave-heated antigenic entity, said entity natively having at least one microwave-resistant epitope, the antigenicity of said microwave treated antigenic entity differing from that of said entity prior to microwave treatment. 17. The process of claim 16 wherein the accessibility of a microwave resistant epitope was increased by the microwave treatment. 18. The process of claim 16 wherein the antigenic entity originally comprised a microwave-labile epitope whose antigenicity was reduced by the microwave treatment. 19. Antibodies prepared by the process of any one of claims 16-18. 20. A non-naturally occurring antigen characterized by the presence of a microwave resistant epitope common to a naturally occurring antigen and the absence of a microwave-labile epitope common to the naturally- occurring antigen, said naturally occurring antigen being associated with an infectious agent. 21. The antigen of claim 20, said antigen being essentially a fragment of said naturally occurring antigen. 22. The antigen of claim 20, said microwave-resistant epitope being cross-reactive with an epitope found on a naturally occurring antigen associated with a normal uninfected vertebrate cell. 23. The process of claim 4 wherein the microwave-resistant epitope is cross-reactive with an epitope found on a naturally occurring antigen associated with a normal uninfected vertebrate cell. 24. In an immunoassay for the detection in a sample of an antigen associated with an infectious agent, wherein an antibody reagent which recognizes said antigen is incubated with the sample or a portion thereof, the improvement consisting of providing an antibody reagent specific for a microwave-resistant epitope of such antigen and of subjecting the sample of a portion thereof to microwave-treatment prior to incubating it with said antibody. 25. In an immunoassay for the detection in a sample of an antibody produced in a subject in response to an antigen associated with an infectious agent, wherein an antigenic reagent recognized by said antibody is incubated with the sample or a portion thereof, the improvement consisting or providing a first antigenic reagent which presents essentially only microwave-resistant epitopes of said antigen. 26. The immunoassay of claim 21 wherein a second antigenic reagent which presents both microwave-resistant and microwave-labile epitopes of said antigen is provided and the assay is run in tandem to compare the amount of antibody immunologically reactive with the first antigenic reagent with the amount of antibody immunologically reactive with the second antigenic reagent. 27. A diagnostic agent comprising an antigen according to any one of claims 15 or 20-22 or an antibody according to claims 13, 14 or 19, in immobilized or labeled form. 28. A therapeutic or prophylactic composition comprising an antigen according to any one of claims 15 or 20-22 or an antibody according to claims 13, 14 or 19, and a pharmaceutically acceptable carrier. 29. A method of inducing immunity in a host to a condition associated with an antigenic entity comprising administering to said host an immunity inducing amount of a composition according to claim 28. 30. A method for the in vitro detection and diagnosis of an antigenic entity having at least one microwave-resistant epitope in an animal or human suspected of having the antigenic entity, which method comprises: contacting a sample from said animal or human with antibodies raised against a altered antigenic entity prepared by irradiating the antigenic entity with microwave radiation of an intensity, wavelength and duration effective to produce the altered antigenic entity which retains said microwave-resistant epitope and provides increased accessibility to said epitope by antibodies which recognize said epitope; and determining whether said antibodies have bound to the antigenic entity. 31. A method for the in vitro detection and diagnosis of an antigenic entity having at least one microwave-resistant epitope and at least one microwave-labile epitope in an animal or human suspected of having the antigenic entity, which method comprises: contacting a sample from said animal or human with antibodies raised against a altered antigenic entity having at least one microwave-resistant epitope prepared by irradiating an antigenic entity having at least one microwave-resistant epitope and at least one microwave-labile epitope antigenic entity, said altered antigenic entity being characterized by the retention of said microwave-resistant epitope and the reduction in the antigenicity of said microwave-labile epitope; and determining whether said antibodies have bound to the antigenic entity. 32. The method of claim 30 or 31 wherein prior to said contacting the sample is irradiated with microwave radiation of an intensity, wavelength and duration effective to produce the altered antigenic entity when said antigenic entity is present in said fluid sample or tissue sample. 33. A method for the in vitro detection and diagnosis of an antigenic entity having at least one microwave-resistant epitope in an animal or human suspected of having the antigenic entity, which method comprises: contacting a sample from said animal or human with antibodies raised against a non-naturally occurring antigen characterized by the presence of a microwave-resistant epitope common to the antigenic entity; and determining whether said antibodies have bound to the antigenic entity. 34. The method of any one of claims 1-3 wherein the antigenic entity is. irradiated with microwaves in the presence of a heat sink so the changes in antigenicity are primarily attributable to nonthermal effects of the microwave treatment. 35. An anti-idiotypic antibody specific for an antibody according to claims 13, 14 or 19.

Inventors: BUNDESEN PETER GREGORY, HILLYARD CARMEL JUDITH, RYLATT DENNIS BRIAN, BUNDESEN, PETER, GREGORY, HILLYARD, CARMEL, JUDITH, RYLATT, DENNIS, BRIAN
Applicants: AGEN LTD, AGEN LIMITED
Application year: 1990
Main classification: G01N33
IPC codes: A61K39, A61K41, C07K1, C07K14, C07K16, C12P21