MEDICAMENTS AND METHODS FOR TREATING AIDS AND ARC EMPLOYING ANTI-CARBOHYDRATE ANTIBODIES AND CARBOHYDRATE ANTIGENS

Abstract: A medicament for inhibiting or slowing the progression of acquired immune deficiency syndrome (AIDS) and AIDS related complex (ARC) comprising a pharmaceutically effective amount of one or more antibodies selected from the group consisting of anti-Le<Y>, anti-AI and anti-sialyl-Tn, and a pharmaceutically acceptable carrier, diluent or excipient. A method for inhibiting or slowing the progression of AIDS and ARC comprising administering to a subject having AIDS or ARC a medicament comprising a pharmaceutically effective amount of one or more antibodies selected from the group consisting of anti-Le<Y>, anti-AI and anti-sialyl-Tn and a pharmaceutically acceptable carrier, diluent or excipient. A vaccine against AIDS and ARC, comprising an immunologically effective amount of one or more antigens selected from the group consisting of Le<Y>, AI and sialyl-Tn, and a pharmaceutically acceptable carrier, diluent or excipient. A method of actively immunizing against AIDS and ARC, comprising administering to a subject a medicament comprising an immunologically effective amount of one or more antigens selected from the group consisting of Le<Y>, AI and sialyl-Tn, and a pharmaceutically acceptable carrier, diluent or excipient.

Description: MEDICAMENTS AND METHODS FOR TREATING AIDS AND ARC EMPLOYING ANTI-CARBOHYDRATE ANTIBODIES AND CARBOHYDRATE ANTIGENS Field of the Invention The present invention relates to treatment of acquired immune deficiency syndrome (AIDS) and AIDS related complex (ARC) , and in particular the present invention relates to medicaments and methods for treating AIDS•and ARC employing the anti-carbohydrate antibodies Le y , j and sialyl-Tn and employing the carbohydrate antigens Le y , A, and sialyl-Tn. BACKGROUND OF THE INVENTION Altered glycosylation in host cells associated with viral infection has been reported (Ray, E.K., et. al. (1978), Virology 88:118; Kumar samy, R., βt. al. (1985), ATCh. Biochβm. BJOPhva. 236:593.. Like oncogenesis, aberrant glycosylation induced by cytomegalovirus or by HIV causes formation of new antigens which are absent in the original host cells (Andrews, P. . βt. al. (1989) J. Exp. Med. 169:1347; Adachi, M., βt. al. (1988) J. Exp. Med. 167:323). Using monoclonal antibodies which define oligosaccharide epitopes, appearance of Le* and Le y antigens after viral infection has been detected. Several studies have indicated the involvement of the carbohydrate part of HIV infection in vitro. Thus, inhibition of the early steps in Golgi glycosylation in infected cells reduces the infectivity of the virus produced (Grutβrs, R.A., βt. al. (1987) Nature 330:74; Montefiori, D.C., βt. al. (1988) Proc. Natl. Acad. Sci. USA 85:9248) . and lectins block syncytium formation probably by a specific interaction with gpl20-glycans of infected cells and also neutralize infectivity of cell-free virus (Lifson, J., et. al. (1986) J. Exp. Med. 164:2101) . Variation in N-glycosylation of target T-4 cells, however, does not seem to influence HIV infection (Montefiori, D.C., et. al. (1988) Proc. Natl. Acad. Sci. USA 85:9248). Gpl20 contains several different glycan structures, and carbohydrate constitutes 50% of the total mass of gpl20 (Matthews, T.J., et. al. (1987) Proc. Natl. Acad. Sci. USA 84:5424) . So far, only N-linked glycans have been found on gpl20 (Kozarsky, K., et. al. (1989) J. Acσ. τmm_ Def. Syndr. 2:163). The binding site on gpl20 for the T4 receptor seems to be located in a non-linear C-terminal part of the molecule (L sky, L.A., et. al. (1987) Cell 50:975) . Whether glycan(s) participate directly in virus-binding is not clear. Thus, inhibitory lectins may bind to glycans adjacent to the binding site and thereby sterically interfere with T4-gpl20 binding as has been found for neutralizing antibodies (Bahraoui, E., et. al. (1988) AIDS 2:165-169; Linsley, P.S., et. al. (1988) J. Virol. 62:3695.. Glycans so far identified on gpl20 by lectin studies are rather ubiquitous, and the therapeutic potential of lectin-based treatment therefore seems small. The present invention is particularly directed to acquired immune deficiency syndrome (AIDS) which is recognized as a distinct new disease whose etiology has been identified as being associated with infection of a new class of lymphotrophic retrovirus termed human immunodeficiency virus (HIV) (previously called, in various terms, human T cell lymphotropic virus III (HTLV-III) , human lymphadenopathy-associated virus (LAV), or AIDS-related virus (ARV) ) (Gallo, R.C. et. al., (1986) Proσ. Allergy. 37:1; Montagnier, L. (1986) Allerαv. 37:46) . The disease is characterized by a disorder associated with an impaired cell- mediated immunity and absolute lymphopenia, particularly reduced helper T lymphocytes (T4 + or CD4) . This is due to the fact that HIV preferentially infects the CD4 lymphocyte population. AIDS may be preceded by a presyndrome that is usually manifested by a complex of designated clinical features and helper T lymphopenia (Friβdman-Kien, A.E., et. al., (1982) Ann, int. Med. 96:693; Gottlieb, M.S., et. al., (1981) N. Enαl. j. Med. 305:1425; Masur, H., βt. al., (1981) M. Bngl. J. Med. 305:1431). The presyndrome is called AIDS-related Complex (ARC) . Diagnosis of infection with HIV is usually made on the basis of detecting antibodies directed against HIV. The exact antibody profile may vary with the stage of the disease (Gallo, R.C. βt. al., (1986) Prog. Allergy. 37:1). Despite significant progress in understanding the pathogenesis of HIV, no studies have been performed regarding carbohydrate antigens in T lymphoid cells infected with HIV, although carbohydrate changes associated with functional change of cellular phenotypes have been well documented (Hakomori, S. (1981) Annu. Rev. Biochem. 50:733) . Evidence has also been developed that the Le y determinant is highly expressed at the surface of human T cell lines only after infection with HIV, as well as in T lymphocytes of peripheral blood of patients with AIDS and ARC but not in normal lymphocytes of healthy individuals (Adachi, M., et. al., (1988) J. Expt. Med. 167:323). Despite the significant advances that have been made to characterize AIDS and ARC, methods of treating and preventing AIDS and ARC are still poorly developed and a great need exists to develop better methods of treatment and prevention of this disease and its presyndrome. SUMMARY OF THE INVENTION Accordingly, one object of the present invention is to find monoclonal antibodies that inhibit in vitro HIV infectively, either by binding to virus or to the target cells used. Another object of the present invention is to use the monoclonal antibodies found to inhibit in vitro HIV infection to define carbohydrate structures expressed as viral glycoproteins and glycosphingolipids associated with the virus, thereby identifying glycans of the viral capsule as targets that are expected to be useful for immuno-therapy and/or vaccine development. These and other objects have been achieved by providing a medicament for inhibiting or slowing the progression of acquired immune deficiency syndrome (AIDS) and AIDS related complex (ARC) comprising: (A) A pharmaceutically effective amount of one or more antibodies selected from the group consisting of anti-Le y , anti-A- and anti-sialyl-Tn, and (B) A pharmaceutically acceptable carrier, diluent or excipient. The present invention also provides a method for inhibiting or slowing the progression of AIDS and ARC comprising administering to a subject having AIDS or ARC a medicament comprising: (A) A pharmaceutically effective amount of one or more antibodies selected from the group consisting of anti-Le y , anti-A j and anti-sialyl-Tn, and (B) A pharmaceutically acceptable carrier, diluent or excipient. In another embodiment, the present invention provides a vaccine against AIDS and ARC, comprising: (A) An immunologically effective amount of one or more antigens selected from the group consisting of Le y , A j and sialyl-Tn, and (B) A pharmaceutically acceptable carrier, diluent or excipient. The present invention further provides a method of actively immunizing against AIDS and ARC, comprising administering to a subject a medicament comprising: (A) An immunologically effective amount of one or more antigens selected from the group consisting of Le y , A j and sialyl-Tn, and (B) A pharmaceutically acceptable carrier, diluent or excipient. In preferred embodiments, the anti-Le y , anti-A, and anti-sialyl-Tn antibodies are monoclonal antibody BMl produced by hybridoma BMl having ATCC deposit no. HB 10312, monoclonal antibody AH21 produced by hybridoma AH21 having ATCC deposit No. HB 10226, and monoclonal antibody TKH2 produced by hybridoma TKH2 (BM4) having ATCC deposit No. HB 9654. BRIEF DESCRIPTION OF THE FIGURES Fig. l is a bar graph showing the inhibitory effect of various monoclonal antibodies on HIV infection of cells. The ordinate represents antigen production, in arbitrary units, and the abscissa represents control with untreated virus (÷AB) , control without virus (÷HIV) , or hybridoma supernatant containing the designated monoclonal antibodies. Figs. 2A to 2D are bar graphs showing inhibition of HIV by various monoclonal antibodies. Fig. 2A: anti-A j monoclonal antibody (MAb) AH21; Fig. 2B: anti- Le y MAb BMl; Fig. 2C: anti-sialyl-Tn MAb TKH2; Fig. 2D: anti-sialyl-Tn MAb B72.3. In Figs. 2A to 2D, the ordinate represents HIV antigen production (% of control) , and the abscissa represents the concentration of monoclonal antibody relative to virus inoculum (μg/TCID 50 ) . DETAILED DESCRIPTION OF THE INVENTION Carbohydrate structures are often involved in initial adhesion of pathogens to target cells. In developing the present invention, a panel of anti- carbohydrate monoclonal antibodies was tested for their ability to inhibit in vitro HIV infectivity, either by binding to the virus or the target cells used. Monoclonal antibodies to three different carbohydrate antigens, i.e., Le y , A- and sialyl-Tn, were able to block infection by cell virus as well as inhibit syncytium formation. The inhibition of virus infectivity was independent of virus strain (HTLV m B and a patient isolate SSI-002) , cell line used for virus propagation (H9 and MT4) or cell type used as infection target (MT4, PMC and T4 lymphocytes). Inhibition was observed when monoclonal antibodies were preincubated with virus but not when cells were so preincubated before infection. The monoclonal antibodies thus are believed to define carbohydrate structures expressed as viral glycoproteins, and glycans of the viral envelope are expected to be useful targets for immuno-therapy and/or vaccine development. Accordingly, the present invention provides a medicament for inhibiting or slowing the progression of acquired immune deficiency syndrome (AIDS) and AIDs related complex (ARC) comprising: (A) A pharmaceutically effective amount of one or more antibodies selected from the group consisting of anti-Le y , anti-A j and anti-sialyl-Tn, and (B) A pharmaceutically acceptable carrier, diluent or excipient. Similarly, the present invention provides a method for inhibiting or slowing the progression of AIDS and ARC comprising administering to a subject having AIDS or ARC a medicament comprising: (A) A pharmaceutically effective amount of one or more antibodies selected from the group consisting of anti-Le y , anti-A j and anti-sialyl-Tn and (B) A pharmaceutically acceptable carrier, diluent or excipient. As the anti-Le y antibody, any antibody can be used as long as it is specific for the carbohydrate antigen Le y having the following structure: Galβl→4GlσNAcβl→R 2 3 t t Fucαl Fuccϋl wherein R represents ceramide or a carrier molecule. Preferred antibodies specific to the carbohydrate antigen Le y have an affinity for Le y corresponding to a KD of about 5x10 *7 . An especially preferred monoclonal antibody specific to the carbohydrate antigen Le y is BMl produced by hybridoma BMl having ATCC Deposit No. HB 10312, and any other monoclonal antibodies having the identifying characteristics of monoclonal antibody BMl are preferred. Monoclonal antibody BMl has the following identifying characteristics: (1) has a KD of about 10 "8 ; (2) isotype is IgM; and (3) reacts with all types of Le y antigens irrespective of length of carrier chains or internal substitution. Additionally, monoclonal antibody BMl is a murine monoclonal antibody. Polyclonal antibodies to the carbohydrate antigen Le y can be prepared by known methods using the purified antigens described in Abe, K., et al (1983) Biol. Chβm. 258:11793. Monoclonal antibodies to Le y can also be prepared by methods known in the art, such as, for example, by the method described by Abe, K., et. al., (1983) J. Biol. Chem. 253:1179. Monoclonal antibody BMl can be prepared by methods known in the art using human gastric cancer cell line MKN 74 as the immunogen. As the antibody to carbohydrate antigen A-, any antibody can be used as long as it is specific to the carbohydrate antigen A j having the following structure: GalNAcαl \ 3 GalBl→3GlcNAcβl→R 2 Fucαl wherein R represents ceramide or a carrier molecule. Preferred antibodies specific to the carbohydrate antigen A j have the following further identifying characteristics: (1) have an affinity for A j corresponding to a KD of about 10 *8 to 5xl0 '7 ; and (2) binds to at least type 1 chain A. Examples of such preferred monoclonal antibodies specific to the carbohydratic antigen j include monoclonal antibody AH21 produced by hybridoma AH21 having ATCC deposit No. HB 10226, monoclonal antibody AH16 (Abe, K. et. al. , ( 1984 ) .T. Timwitial . 132 : 1951) and any other monoclonal antibodies having the identifying characteristics of either of these monoclonal antibodies. Monoclonal antibody AH21 is especially preferred. Monoclonal antibody AH21 has the following identifying characteristics: (1) has a KD of about 5xl0 "7 ; (2) isotype is IgM; and (3) specifically binds to type 1 chain A. Additionally, monoclonal antibody AH21 is a murine monoclonal antibody. Monoclonal antibody AH16 has the following identifying characteristics: (1) has a KD of about 10 "8 ; (2) isotype is IgG 3 ; and (3) binds to all types of A antigens, including type 1 chain A, irrespective of carrier carbohydrates or internal substitution. Additional characteristics of monoclonal antibody AH16 are described by Abe, K. et. al., (1984) j_ Immunol. 132:1951. Polyclonal antibodies to the carbohydrate antigen A- can be prepared by known methods, using the purified antigens described in Abe, K., et. al., (1983) J. Immunol. 132:1951. Monoclonal antibodies to the carbohydrate antigen A. can be prepared by methods known in the art, such as, for example, by the method described in Abe, K., et. al., (1984) .T- TBur nr>t . 132:1951. Monoclonal antibody AH21 can be prepared by methods known in the art using purified type 1 chain A as the im unogen. Monoclonal antibody AH16 can be prepared by methods known in the art using human gastric cancer cell lines as the immunogen. As the anti-sialyl-Tn antibody, any antibody can be used as long as it is specific for the sialyl-Tn carbohydrate antigen having the following structure: NeuAcα2→6GalNAcαl→0-Ser/Thr wherein Ser represents serine and Thr represents threonine. The isotypes and affinities of suitable anti- sialyl-Tn antibodies for the antigen vary extensively from one antibody to the next, and thus the specificity to sialyl-Tn is the critical identifying characteristic of these antibodies. Examples of preferred monoclonal antibodies include TKH2 produced by hybridoma TKH2, having ATCC Deposit No. HB 9654, TKH1 (Kjeldsen, T. et. al., (1988) Cancer Res. 48:2214-2220), B72.3 (Colcher, D. et. al., (1981) Proc. Natl. Acad. Sci. USA. 78:3199- 3203) , and any other monoclonal antibodies having the identifying characteristics of these monoclonal antibodies. Monoclonal antibody TKH2 is especially preferred. In addition to binding specifically to the carbohydrate antigen sialyl-Tn, a further identifying characteristic of monoclonal antibodies TKH1, TKH2 and B72.3 is their isotypes which are IgM, IgG- and IgG-, respectively. Further, monoclonal antibody TKH2 is a murine monoclonal antibody. Additional characteristics of monoclonal antibodies TKH1 and B72.3 are described in Kjeldsen, T. et. al., (1988) Cancer Res. 48:2214-2220 and Colcher, D. et al., (1981) Proc. Natl. Acad. Sci. USA. 78:3199-3203, respectively. Polyclonal antibodies to the carbohydrate antigen sialyl-Tn can be prepared by known methods, Kjeldsen, T. et. al., (1988) Cancer Res. 48:2214. Monoclonal antibodies to the carbohydrate antigen sialyl-Tn can be prepared by methods known in the art, such as those described in Kjeldsen, T., et. al., (1988) Cancer Res. 48:2214 and Nuti, M., et. al., (1982) J. nst. Cancer 29:539. Monoclonal antibodies to sialyl-Tn can also be prepared by the following novel method disclosed in co-pending application U.S. Serial No. 317,492, the entire disclosure of which is incorporated herein by reference. The immunogen used to obtain the antibody to sialyl-Tn antigen is the sialyl-Tn antigen which is a core structure of many mucin-type glycoproteins. Mucin-type glycoprotein, as used herein, means a high molecular weight protein (Mr>10 6 ) with a high degree of O-linked glycosylation at serine or theronine residues. Mucin-type glycoproteins are further polymerized by S-S-dependent linkage and are the major components of epithelial secretions. Also, core structure of mucin-type glycoprotein, as used herein means a basic carbohydrate structure without peripheral substitution and which is directly linked to the protein moiety of a mucin-type glycoprotein. In the present invention, any sialyl-Tn core structure of a mucin-type glycoprotein can be used as an immunogen as long as the glycoprotein has a high molecular weight (relative molecular weight > 10 6 daltons) and is glycosylated to the same degree as mucin, i.e., more than 50% of total weight is glycosylated. Animal mucins containing the sialyl- Tn antigen can also be used as an immunogen. The sialyl-Tn antigen immunogen can be obtained by enzymatic or chemical modification of a mucin-type glycoprotein to expose the sialyl-Tn antigen core structure or by isolation of mucins having on them sialyl-Tn core structures. These mucins are present in some animal species. Examples of types of enzymatic modifications that can be used to expose the sialyl-Tn core structure of various mucin-type glycoproteins include the elimination of the terminally located α2→3 sialyl residue by influenza virus sialidase or the total elimination of all sialic acid residues by Clostridium perfrinαense sialidase. Enzymatic modification can also include treatment with β-galactosidase (preferably from Charonia lampas) , α-fucosidase, and N-acetylhexosaminidase. Enzymatic hydrolysis of mucin glycoprotein is described by Hirohashi et al. (Proc. Nat. Acad. Sci. USA (1985) 82_:7039-7043) . Examples of chemical reactions which can be used to expose the sialyl-Tn core structure of mucin-type glycoproteins include periodate oxidation followed by reduction with sodium borohydride and treatment with weak acid. The procedure is called Smith degradation (Sprio. G., Methods Enzvmol. (1972) 28:3-43). This chemical treatment eliminates non-reducing terminals of carbohydrate residues except sialic acid, which can be eliminated by sialidase treatment, as described above. Examples of mucins isolated from animals that can be used as immungens include ovine submaxillary mucin (OSM) in which 90% of the carbohydrate chains consist of the sialyl-Tn antigen and bovine submaxillary mucin (BSM) in which 50% of the carbohydrate chains consist of the sialyl-Tn antigen and 20% of the carbohydrate chains consist of T antigen and other unidentified residues. The sialyl-Tn antigen immunogen is isolated and purified according to conventional methods. For example, mucin-type glycoproteins which will be enzymatically or chemically modified to produce a sialyl-Tn core structure can be isolated by gel filtration through Sepharose 4B or Sephacryl 200S. The isolated glycoprotein is then enzymatically or chemically modified, by methods described above, to expose the sialyl-Tn core structure, and the sialyl- Tn core structure is purified for use as immunogen as follows. The modified mucin can be separated by gel filtration through Sepharose 4B or Sephacryl 200. High pressure chromatography on a synthetic molecular filter column (fast liquid chromatography; Pharmacia) is also useful to separate enzymatically or chemically modified mucins. However, as immunogen for making monoclonal antibodies, the modified mucin does not need to be purified. The presence of a small quantity of unmodified mucin will not be harmful to use as an immunogen according to the present invention. Further, modification is usually quantitative, if proper routine precautions are taken. Mucins which are derived from animal species and contain glycoproteins already in the form of a sialyl- Tn core structure are obtained by conventional methods. For example, by gel filtration through Sepharose 4B, Sephacryl 200, or FPLC, as described above. The thus derived animal mucins can be further purified for use as immunogens by the same methods, such as conventional gel filtration or FPLC as described above. However, the purity of the immunogen is not essential for use to produce a monoclonal antibody. Human erythrocyte glycophorin can be obtained from human erythrocyte membranes by the method originally described by Marchesi, V.T. and Andrews, E.D. (Science (1971) 174.:1247-1248) . Whether a glycoprotein has the sialyl-Tn core structure can be determined by affinity chromatography with a peanut lectin column (Carter, W.G., and Sharon, N., Arch. Biochem. Biophys.. (1977) 180:570- 582) or by immunoblotting of glycoproteins with anti- Tn antibody (available from ChemBiomed, Edmonston, Alberta, Canada) . The distribution of the sialyl-Tn antigen is rather limited. However, one source of the sialyl- Tn antigen which is useful in the present invention is culture supernatants of squamous lung carcinoma cell lines QG 56 and LU-65 (Hirohashi S, et. al., (1985) Proc. Natl. Acad. Sci. USA. 82:7039-7043). A second source is again ovine submaxillary mucin, which contains a high density of sialyl-Tn. In order to obtain sialyl-Tn antigen in a form useful as the immunogen for the present invention from culture supernatant, the various cell lines described above are cultured according to known methods. The culture supernatants are treated to obtain sialyl-Tn antigen as follows. The spent culture medium from cells cultured in suspension is lyophilized to reduce its volume to 1/50 of the original volume. The concentrated spent medium is then dialyzed extensively against phosphate- buffered saline containing 0.01% sodium azide at 4 β C. The dialyzed material is placed on Sepharose 4B and gel filtrated. The void volume is pooled, concentrated further, and re-chromatographed on Sephacryl 200. The glycoprotein fraction in the void volume is used as immunogen. All procedures must be completed within a limited time and at low temperature (4°C) , since the εialyl linkage is unstable. The immunogen, prepared as described above, is treated for immunization as follows. The sialyl-Tn antigen (e.g., 4.0 mg) is dissolved in distilled water (e.g., 4 ml) , thoroughly mixed with an appropriate amount of acid-treated Salmonella minnesota (e.g., 16 mg) , and lyophilized. The dried mixture is suspended in a suitable volume (e.g., 4.0 ml) of an appropriate carrier (e.g., 140 mM NaCl containing 20 mM phosphate buffer, pH 7.0), and aliquots of about 100 μg of mucin and 400 μg of bacteria are injected intravenously. Immunization can also be made with complete Freund's adjuvant instead of adsorption on bacteria, and the ratio between the amount of mucin and Salmonella minnesota can be varied. The best results have been observed when Salmonella minnesota is treated with acetic acid, as previously described (Young, W.W., et. al., (1979) J.EXP. Med. 150:1008- 1019) . The host used for immunization can be a mouse or rat of any strain or any other type of animal whose splenocytes are suitable for preparation of hybridomas, i.e., susceptible to cell fusion with HAT- sensitive myeloma cell lines to establish stable hybridomas. The immunization schedule depends upon the host animal susceptibility to mucin immunization, but the protocol described above is suitable for mice. Alternative conditions can also be applied. Suitable immunization ' schedules can be determined by the skilled artisan. For example, a suitable immunization schedule for Balb/c mice is to inject the immunogen preparation intravenously through the caudal vein once a week for 5 weeks and then after a one-month intermission, boost with the immunogen preparation. The amount of immunogen preparation administered to the host depends upon the molecular weight of the mucin, the exposure of the carbohydrate epitope, and the novelty and density of the epitope associated with mucin-type glycoproteins. The range of glycoprotein injected in mice is 3-5 μg coated on 30-50 μg of Salmonella minnesota suspended in 100 μl of saline, intravenously injected in each individual mouse, whose body weight range is 100-150 g. When complete Freund's adjuvant is used, about 20 μg of glycoprotein in 500 μl of saline is emulsified with 500 μl of complete Freund's adjuvant, and about 200 μl is injected subcutaneously at multiple sites (about 50 μl per site) . Similar quantities of antigen, either coated on Salmonella minnesota or mixed with complete Freund's adjuvant, are used for other hosts such as rats, hamsters, or guinea pigs, and several times greater quantities are used for other hosts such as rabbits. It is not necessary to proportionately increase the amount of antigen to the body weight of the animal. Immunization is repeated until sufficient antibody is detectable in whole serum. The spleen cells of the host are removed and splenocytes are fused with HAT-sensitive myeloma cells by the technique that has been well established (Kδhler, G. and Milstein, c, (1975) Nature 256:495-497 and Young, . ., et. al., supra) . HAT-sensitive myeloma cells can be NS-1, SP-1 or SP-2, for example, but any type of HAT-sensitive myeloma cells can be used. Occasionally, hybridomas can be established after fusion of host splencytes with myeloma cells even though no antibody was detectable in the host serum. Therefore, it is not essential to detect antibodies before cell fusion. Fusions are usually carried out in polyethylene glycol, as detailed by Young, . ., et. al. (supra). A preferred myeloma cell line for use in the present invention is SP-2. The fused cells are cultured in 96-well plates until miniclones are formed. It is important to use splenocytes 48-72 hours after the last booster injection and to fuse with well proliferating myeloma cells to obtain a number of surviving fused cells to grow. Moisture and C0 2 concentration in the incubator must be carefully controlled at the initial stage of culturing the fused cells. Feeder cells are not necessary to grow the fused cells according to the process of the present invention. One skilled in the art can readily determine suitable culture conditions. After an appropriate culture period, hybridomas secreting antibodies that react with the sialyl-Tn antigen that was used to immunize are cloned and subcloned by limiting dilution, i.e., by diluting to a point where less than one cell per new culture will be expected, and then plating into the wells. In general, each well of a 96-well plate is coated with mucin glycoprotein containing the sialyl- Tn core structure used as immunogen by incubation of each well with glycoprotein solution in PBS, i.e., 5- 10 μg/50 μl/well is added and incubated overnight. Glycoprotein solution is removed, washed, and further incubated with bovine serum albumin (10 μg/50 μl/well) to block the plate before using it to screen antibodies. This method is described by Hirohashi et al. (supra) . The hybridomas that secrete the antibodies that react with the particular antigens are screened as follows. Antibody bound to an antigen-coated well is usually detected by secondary antibody (anti-mouse IgM and IgG goat or rabbit antibodies) followed by 125I- labeled protein A as initially described by Young, W. ., et al. (supra). The method is still more sensitive than available ELISA assays, although an ELISA can also be used. ELISA's can be more conveniently processed by use of automated readers and ELISA kits available commercially. Monoclonal antibodies to sialyl-Tn antigen secreted by hybridomas thus isolated can be produced in quantity by growing large batches of hybridoma cell cultures and purifying the antibody from the supernatant or by injecting mice with the hybridoma line to stimulate the production of ascites fluid. Both methods are well known in the art. Methods of producing the monoclonal antibody in quantity according to the present invention are described in Young et al. (supra) . The hybridomas isolated according to the present invention can be grown in large batches in suspension culture, or more conveniently in a fiberglass container in which cells are packed and grown in high density, wherein antibodies can diffuse into the culture medium. The monoclonal antibodies can be purified by known methods, for example, by affinity separation using protein A or high pressure liquid chromatography on reverse phase alkylated silica gel or with a synthetic polystyrene gel filtration column. Suitable doses of the above-described medicament can readily be determined by the skilled artisan, depending upon the mode of administration. More specifically, a suitable dosage for intravenous administration of antibodies to suppress further AIDS infectivity at the initial stage of patients with AIDS should be determined by various conventional experiments, since TKH2 antibody can inhibit HIV infection at 2 μg/ml concentration in vitro. 15.6 mg of TKH2 is believed sufficient to prevent HIV infectivity when administered to a man of 60 kg weight who has approximately 7.8 1 of blood. Humanized antibodies are highly preferable to perform this in vivo inhibition. Suitable methods of administration of the above- described medicament can also be determined by those skilled in the art. Antibodies can be administered intravenously, however, other forms of administration are also possible. Suitable pharmaceutically acceptable carriers, diluents and excipients are readily determined by the skilled artisan. For example, the antibodies can be administered when solubilized in a physiological buffer solution. However, no carrier is considered especially preferable. The present invention also provides a vaccine against AIDS and ARC, comprising: (A) An immunologically effective amount of one or more antigens selected from the group consisting of Le y , A, and sialyl-Tn, and (B) A pharmaceutically acceptable carrier, diluent or excipient. Similarly, the present invention provides a method of actively immunizing against AIDS and ARC, comprising administering to a subject a medicament comprising: (A) An immunologically effective amount of one or more antigens selected from the group consisting of Le y , A j and sialyl-Tn, and (B) A pharmaceutically acceptable carrier, diluent or excipient. The antigen Le y , whose structure is defined above, can be prepared either synthetically by known methods (Hindsgaul, O. βt. al., (1982) Carbohydrate Res. 109:102-142 and Spohr, U. et. al., (1985) Canadian J. Cham. 63:2644-2652) or, alternatively, can be isolated from naturally occurring sources by known methods, such as described in Abe, K. et. al., (1983) J. Biol. Chem. 258:11793. The antigen A,, having the structure set forth above, can also be prepared synthetically by known methods (see Lemieux, R.U. Chemical Society Reviews (1978) Vol. 7 423-452; IUPAC Frontiers of Chemistry 1982 K.J. Laidler, βd. 3-24 for the synthesis of Type 1 chain A) or can be isolated from natural sources, also by known methods such as that described in Abe, K. et. al., 11980 J. Immunol. 132.1951. The antigen sialyl-Tn, whose structure is set forth above, can be isolated from natural sources by known methods, such as that described in Kjeldsen, T. et. al., (1988) Cancer Res. 48:2214, or by the method described above for preparing purified immunogen comprising the sialyl-Tn antigen. Suitable immunologically effective doses and pharmaceutically acceptable carriers, diluents and excipients of the above-described vaccine can readily be determined by the skilled artisan. For example, an immunologically active dose of sialyl-Tn antigen as a vaccine will be approximately 55 μg per injection. However, the carrier molecule is important. BCG coated with sialyl-Tn antigen is considered to be a useful way to prevent AIDS infectivity. One example is 50 μg of sialyl-Tn antigen coated on 500 μg of BCG injected intradermally or subcutaneously. Suitable methods of administration of the above- described vaccine can also be readily determined by the skilled artisan. EXAMPLES The present invention will now be described by reference to non-limiting examples. Unless otherwise specified, all percents, ratios, parts, etc. are by weight. Materials and Methods for Examples Anti-carbohydrate monoclonal antibodies Antibodies tested, their specificity, isotype, and reference for production, are listed in Table I. The twenty different monoclonal antibodies shown in Table I define carbohydrate structures or glycosphingolipids found at the periphery of both N- linked and O-linked, or exclusively O-linked, chains of glycoproteins and also glycosphingolipids (for a review see Clausen, H., and S. Hakomori (1989) Vox Sang. 56:1) . All cells were grown in RPMI 1640 containing 10% fetal calf serum (FCS) , 2 mM glutamine and 1 mM pyruvate, and stored at 4°C with 0.02% NaN 3 . The study was performed in three stages. Initially, all listed monoclonal antibodies were tested for HIV-inhibition as culture hybridoma supernatants containing about 10-50 μg immunoglobulin (lg)/ml. In the second stage, monoclonal antibodies showing some reduction of HIV infection (TKH2, BMl, AH21, AH16, NKHl, HH6, SH2, CA3F4) were retested also as hybridoma supernatants, but after extensive dialysis of the hybridoma supernatants against PBS to remove preservatives (Example 1; Fig. 1) . Finally, the monoclonal antibodies TKH2, B72.3, BMl and AH21 were used after purification from culture supernatant (Example 2; Figs. 2A to 2D). TKH2 (IgG,) and B72.3 (IgG,,) were purified by protein A-Sepharose chromatography, and AH21 and BMl by ion-exchange chromatography as described by the manufacturer (Pharmacoa, Upsala, Swedan) . TABLE I. Antigen specificity and reference for monoclonal antibodies used in this study. Antigen Antibody Reference Type 1 chain structures Le d - sialyl Le' II / Fucαl Type 2 chain structures N-lac Galβl→4GlcNAcβl→R IB2 (IgM) (4) H Galβl→4GlcNAcβl→R BE2 (IgM) (4) 2 t Fucαl Le x (ext. Le x ) Galβl→4GlcNAcβl→R SHI, -2 (IgG) 3 ♦ Fucαl L β y Galβl→4GlcNAcβl→R BMl (IgM) (5) 2 3 t t Fucαl Fucαl sialyl Le x NeuAcα2→3Galβl→4GlcNAcβl→4Galβl→4GlcNAcβl→R FH6 (IgM) (6) co 3 3 ±Fucαl Fucαl sialyl I NeuAcα2→3Galβl→4GlcNAcβl Galβl→4GlcNAcβl→R NUH2 (IgM) 6 NeuAcα2→3Galβl→4GlcNAcβl NeuAcα2→6Galβl→4GlcNAcβl→3Galβl→4GlcNAcβl→R NUH1 (IgM) 3 t Fucα1 ALe' GalNAcαl Galβl→4GlcNAcβl→R A*"; 6 ' [35 2 3 Fucαl Fucαl T ype 3 chain structures Tn sialyl-Tn O Tα Mil Type 4 chain structures Tβ Galβl→3GalNAcβl→4Galβl→R TKH7 (IgM) Fuc-GM j Galβl→3GalNAcβl→4Galβl→4Glcβl→Cer TKH5 (IgM) 2 3 t t Fucαl NeuAc 2 1. Young, W.W.Jr., et. al. (1983) J. Biol. Cheml 258:4890 2. Nudelman, E., et. al. (1986) J. Biol. Chem. 261:5487 3. Abe, K. , et. al. (1984) J. Immunol. 132:1951 4. Young, W.W. Jr., et. al. (1981) J. Biol. Chem. 256:10967 5. Abe, K., et. al. (1983) J. Biol. Chem. 258:11793 £ 6. Fukushi, Y., et. al. (1984) J. Biol. Chem. 259:10511 7. Hirohashi, S., et. al. (1985) Proc. Natl. Acad. Sci. USA 82:7039 8. Kjeldsen, T. , et. al. (1988) Cancer Res. 48:2214 9. Nuti, M. , et. al. (1982) J. Inst. Cancer 29:539 10. Clausen, H. , et. al. (1988) Molec. Im un. 25:199 11. Clausen, H. , et. al. (1985) Proc. Natl. Acad. Sci. USA 82:1199 R = Ceramide or a carrier molecule Cells For HIV infection experiments, the T4-lymphocyte cell lines H9 (Popovic, M., et. al. (1984) Science 224:497) , CEM (Foley, G.E., et. al. (1965) cancer 18:522) and MT4 (Harada, S., et. al. (1985) Science 229:563) were cultured at 37°C, 5% C0 2 using RPMI 1640 With 10% FCS (5% for CEM cells) , 100 IU/ml penicillin, 20 μg/ml gentamicin and 100 IU/ml streptomycin (growth medium) . Cells were maintained at a concentration of 2-19 x 10 5 cells/ml and medium was exchanged twice weekly. Peripheral blood mononuclear cells (PMC) were obtained from healthy donors by Ficoll-Hypaque gradient centrifugation (Boyum, A., (1968) Scand. J. Clin. Lab. Invest. 21:suppl. 97) and T4-lymphocytes were selected by panning (Wysocki, L.J., et. al. (1978) Proc. Natl. Acad. Sci. USA 75:2844) . Donor PMC and T4-lymphocytes were cultured as above in growth medium supplemented with 20 IU/ml Interleukin-2. Virus Source Supernatant from H9 cells chronically infected with reference HIV-1 strain HTLV j , j B, (Popovic, M., et. al., (1984) Science 224:497), was sterile-filtered, aliquotted and stored at -80°C until use. HIV-1 strain SSI-002 was isolated from an HIV-infeσted patient (CDC II) as previously described (Nara, P.L. et. al., (1989) Proc. Natl. Acad. Sci. USA 86:7139- 43) . The virus was then passed three times in MT4 cells, and stored as above. Before use, the TCID 50 of the virus preparations was determined in MT4 cells and PMC. Toxicitv Toxicity of the monoclonal antibody preparations was examined by incubating 0.5xl0 6 MT4 cells in 24-well cell culture plates in growth medium containing from 20 to 0 μg/ml purified antibody, and supernatants were exchanged with fresh medium containing appropriate concentrations of antibody after 4 days of culture. Live cell counts were obtained on days 0, 4 and 7 using trypan blue exclusion. ELISA Cell-free culture supernatants were examined for HIV antigen using a double antibody sandwich ELISA as described previously (Nielsen, CM., et. al. (1987) Lancet 8532:566). Each plate included a dilution series of a standard HIV antigen preparation, and optical densities (490 nm) were expressed relative to this standard preparation (arbitrary units) . All in vitro infection experiments included a control using untreated HTLV m B. After 4 days of culture, supernatants were diluted to give an OD490 from this control of approximately 2. All antibodies tested for inhibition of infection were also tested for interference with the ELISA detection, and no such interference was found. Polymerase Chain Reaction (PCR) and Dot Blot Hybridization Cell pellets were suspended in 100 μl Triton-X-100 in phosphate-buffered saline (PBS) and boiled for 10 minutes to release DNA. 10 μl was mixed with 90 μl of amplification mixture (200 pmol of each primer, 200 μmol each of dATP, dCTP, dGTP and dTTP, 50 mM KCl, 10 mM Tris (pH 8.3), 6.5 mM MgCl 2 , and 0.02% • gelatin) . The primers used were SK29/SK30 (Ou, C.-Y., et. al. (1988) Science 239:295) . The mixture was heated to 94 β C for 5 minutes and then 2 units Taq- polymerase (Perkin-Elmer Cetus, Norwalk, USA) were added. Forty rounds of amplification were carried out in a DNA Thermal Cycler (Perkin-Elmer Cetus) . Each cycle lasted 1 min at 94°C, 1 min at 65°C and 1 min at 72 β C. One μl of the amplified product was mixed with 60 μl 0.67 M NaOH/0.67 M NaCl and boiled for 5 minutes followed by cooling on ice. Samples were transferred to a Zeta-probe membrane in a vacuum manifold and sample wells were rinsed with 500 μl of 0.4 M NaOH each. The membrane was rinsed briefly in 2 X standard saline citrate solution (SSC; 1 X SSC: 0.15 M NaCl, 0.015 M Na citrate pH 7.0), air dried and heated at 80°C for 2 hours. Membranes were prehybridized for 2 hours at 42°C in hybridization solution (50% deionized formamide, 1% sodium dodecyl sulphate (SDS) , 1 M NaCl, 50 mg/ml dextran sulphate and 0.1 mg/ml sheared salmon sperm DNA) , whereafter 20 pmol of 32 P-labeled probe (SK31) was added and hybridization was performed overnight at 42°C. Membranes were washed 10 minutes in 2 X SSC, 0.1% SDS at room temperature, followed by 30 minutes at 2 X SSC, 0.1% SDS at 62 β C. Finally, the membrane was rinsed 2x15 minutes in 0.2 X SSC, 0.1% SDS at room temperature and exposed to Kodak XAR films overnight. Indirect Immunofluorescence Analysis Binding of the monoclonal antibodies to cells was determined by indirect immunofluorescence microscopy (Inanue, E.R., et. al. (1986) In:D. M. Weir tea) . TmirπiTtacaemistry ι, 23:1). Briefly, 10 6 cells resuspended in 50 μl suspension buffer (PBS containing 1% BSA, 2% human serum and 0.1% azide) were incubated with 50 μl hybridoma supernatant for 45 min at 4°C. The cells were then washed once in 1 ml cold PBS and incubated for another 30 min at 4°C with FITC-conjugated rabbit anti-mouse immunoglobulin (Dakopatts, Copenhagen) diluted 1:30 in suspension buffer. After incubation the cells were washed once, resuspended in 12 μl mounting buffer (PBS containing 30% glycerol, 0.1% azide and 1% paraformaldehyde) and transferred to a slide for microscopy. At least 100 cells were counted and the percentage of fluorescent cells was calculated. Monoclonal antibodies against CD4 (0KT4 from The Hybridoma Laboratory, State Serum Institute, Copenhagen, and MT310 from Dakopatts, Copenhagen) and against gpl20 (DuPont) were used as controls. EXAMPLE 1 Screening for Inhibitory Effect of HIV Infection Eight hybridoma supernatants, containing monoclonal antibodies (NKHl, HH6, TKH2, SH2, BMl, AH21, AH16, and CA3F4) to eight different oligosaccharides, were extensively dialyzed against PBS and sterile filtered (0.22 μm) . Virus inhibition assay. Ten TCID 50 HTLV B was mixed with 0.5 ml hybridoma supernatant and incubated for 1 hour at 37°C. MT4 cells (lxlO 6 ) were suspended in this mixture and incubated for 2 hours at 37°C. After extensive washing the cells were resuspended in growth medium (4ml) containing 10% v/v of the corresponding hybridoma supernatant; and duplicates (1.5 ml) were transferred to a 24 well cell-culture plate. The cells were cultured for 7 days at 37°C, 5% C0 2 , and cell-free supernatant (750 μl) was exchanged with fresh medium without hybridoma supernatant after 2, 4 and 7 days. HIV antigen output was measured by ELISA as described above on the supernatants. Dialyzed hybridoma supernatants (200μl) were preincubated with 10 TCID 50 HTLV ιπ B or SSI-002 before inoculation and cultured as described above. Purified monoclonal antibody was preincubated with 10 TCID 50 HTLV πι B which was then used to inoculate 1 x 10 6 MT4 cells, 4 x 10 6 3-day PHA stimulated PMC or selected T4-lymphocytes. After incubation for 2 hours at 37°C, cells were washed and resuspended in 5 ml growth medium. Quadruplicates of 1 ml cell suspension were transferred to a 24-well cell culture plate. Supernatant (500 μl) was exchanged with fresh growth medium after 4 and 7 days of culture. The results are shown in Fig. 1. In Fig. 1, the ordinate represents antigen production, in arbitrary units, and the abscissa represents control with untreated virus (÷AB) , control without virus (÷HIV) , or hybridoma supernatant containing the designated monoclonal antibodies. Of the monoclonal antibodies listed in Table 1, the following monoclonal antibodies reduced HIV antigen concentration in the culture supernatants to background levels: TKH2, BMl, AH21 and AH16 (having similar but broader specificity than AH21) . Cells from these cultures were analyzed for HIV-DNA by polymerase chain reaction after 4 days of culture. All cultures with HIV antigen negative supernatants (TKH2, BMl, AH21, AH16 and the uninfected control) were also negative for HIV-DNA, and cultures with antigen positive supernatants were also positive for HIV-DNA. EXAMPLE 2 Inhibition of Infection in Different Target Cells In order to compare the inhibitory effect of the monoclonal antibodies on HIV infection in different cell systems, cells of the MT4 cell line, PHA-stimulated PMC and selected T4-cells from a normal donor were used as target cells for infection with HTLV JJJ B preincubated with the monoclonal antibodies AH21, TKH2, B72.3 and BMl. 150 TCID 50 HTLV JJJ B was incubated in a total volume of 300 μl RPMI 1640 containing 6 μg affinity-purified monoclonal antibody for l hr at 37 β C. 20 μl (= 10 TCID 50 for MT4 cells) was added to 2xl0 6 MT4 cells or 100 μl (= 10 TCID 50 for normal lymphoblasts) was added to 4xl0 6 3-day PHA-simulated donor lymphoblasts or panned T4 lymphocytes, and incubated for 2 hrs at 37°C. After washing, the cells were resuspended in 5 ml growth medium containing 1 μg/ml antibody and for lymphoblasts also 20 IE/ml interleukin-2, and 4x1 ml was plated in a 24-well cell culture plate. 500 μl supernatant was exchanged with fresh growth medium after 4 and 7 days of culture. The supernatants were stored at -20°C until use in ELISA as described above. Determination of inhibitory concentration 20 TCID 50 HTLVJJ J B was incubated in a total volume of 60 μl RPMI 1640 containing affinity-purified monoclonal antibodies in varying concentrations for 1 hr at 37°C. 30 μl (= 10 TCID 50 ) was added to lxlO 6 MT4 cells in 500 μl RPMI, and incubated for 2 hrs at 37°C. After extensive washing, the cells were resuspended in 5 ml growth medium containing 1/10 of the antibody concentration used for HIV preincubation, and 4x1 ml cell suspension was plated in a 24-well cell culture plate. 500 μl supernatant was exchanged with fresh growth medium after 4 and 7 days of culture using Trypan blue. The supernatants were stored at -20°C until use in ELISA as described above. The results are shown in Figs. 2A to 2D. In Figs. 2A to 2D, the ordinate represents HIV antigen production (% of control) , and the abscissa represents the concentration of monoclonal antibody relative to virus inoculum (μg/ICID 50 ) . Figs. 2A to 2D show that purified antibodies of hybridoma culture supernatant of AH21, BMl, TKH2, and as a control B72.3 (having the same specificity as TKH2) showed a concentration dependent inhibition of infection. The inhibitory antibody concentration was defined as the antibody concentration resulting in an 80% reduction of HIV antigen expression after 4 days of culture, as described for neutralizing monoclonal antibody (MAb) titers (Kinney Thomas, E., et. al. (1988) AIDS 2:25) , and expressed relative to infectious dose. The inhibitory concentrations were interpolated to be: B72.3:0.18 μg/TCID 50 ; AH21: 0.32 μg/TCID 50 ; BMl: 1.16 μg/TCID 50 ; TKH2: 1.6 μg/TCID 50 . The monoclonal antibodies inhibited infection with both HIV-1 reference strain HTLV JJJ B and isolate SSI-002 when the virus preparation was preincubated with MAb before inoculation onto MT4 cells. No reduction in HIV infection was found when the cells were preincubated with MAb and then inoculated with untreated virus. Cell immunofluorescence A cell immunofluorescence assay was conducted as described above. Table 2 summarizes staining results with the selected monoclonal antibodies. AH21 and AH16 did not label cells. TKH2 weakly labeled a small population of cells, whereas B72.3 did not stain cells. Monoclonal antibodies BMl labeled uninfected CEM and H9 cells, as well as infected H9 cells. BMl did not stain MT4 cells, uninfected or infected, and only a small percentage of T4 lymphocytes purified from a normal donor by panning were positive. TABLE II. Indirect immunofluorescence staining* of various types of T cell lineage, uninfected or infected with HIV-1 strains HTLV-III B or SSI-002, using carbohydrate-specific Monoclonal Antibodies, anti-gpl20 and anti-CD4. ♦Results expressed as percentage of cells labeled. ND, not determined. Host cell glycolipids may be incorporated in virions (Klenk. H.-D., et. al. (1970) Proc. Natl. Acad. Sci. USA 66:57), which should thus show a glycosylation pattern similar to the host cell. The glycosylation of viral proteins is also believed to be performed by the host cell gene-encoded glycosyltransferases. Therefore, it is generally expected that the host cell and proteins of viruses propagated therein show similar glycosylation patterns. However, the final glycan structure of a glycoprotein is a result of both exoglycosidase and glycosyltransferase activity as well as primary structure of the core protein, which in the case of viral glycoproteins could result in novel glycosylation patterns. Alternatively, virus infection could induce expression of abnormal glycosyltransferase genes in the host cell. The present results showed no correlation between inhibition of infection and immunofluorescence staining of cells used as targets or as virus source. The virus inhibition of the MAbs was thus independent of cell type used for virus propagation. EXAMPLE 3 Inhibition of svncvtium formation Two-fold dilutions of HTLV JJJ B infected H9 cells (10 5 -10 2 cells) in growth medium (25 μl) were mixed with dialyzed hybridoma supernatant (25 μl) in a 96-well microtiter plate and incubated for 1 hour. Then 2xl0 5 CEM cells in growth medium (50 μl) were added. After incubation for 24 hours, cultures were evaluated for syncytia by microscopy. Syncytial activity was defined as the minimal number of infected H9 cells where syncytium formation (giant-cells were more than 5 nuclei) was detectable in the cultures. The results are shown in Table III. Table III Reduction of syncytial activity of carbohydrate specific monoclonal antibodies after preincubation of infected H9 cells. Syncytial activity expressed as minimal number of HTLV jjj B infected cells where syncytium formation was detectable after 24 hours coincubation with CEM cells. Antibody Number of infected cells None (control) 3,125 B72.3 12,500 BMl 12,500 AH21 6,250 TKH2 >100,000 (no syncytia) The results indicate that the identified carbohydrate antigens are expected to function as targets for passive immunotherapy using the appropriate monoclonal antibodies. Also, the identified antigen may be used to provoke an immune response in patients by active immunization. In the present study, a panel of murine monoclonal antibodies was employed defining peripheral carbohydrate structures (generally 2-5 residues) which may be found on poly-N-acetyllacetosamine structures of both N- and O-linked glycoprotein and glycospingolipid types, as well as structures restricted to O-linked glycoprotein types (Table I) . Monoclonal antibody BMl, directed to Le y and previously found to be expressed on HIV-infected lymphocytes (Adachi, M., et. al. (1988) J. Earn. Med. 167:323, PincUS, S.H., et. al. (1989) .T_ Tnιιmιτ.r.1. 142:3070), inhibited formation of syncytia between infected and uninfected cells and also the infectivity of cell-free virus. This inhibition was observed only on preincubation of monoclonal antibody with virus, but not on preincubation of monoclonal antibody with cells, before inoculation. One monoclonal antibody, AH21 (IgM) , with specificity for blood group A type 1 chain antigen and one monoclonal antibody, AH16 (IgG 3 ) , that binds to all types of A antigens, including type 1, irrespective of carrier carbohydrate or internal structure, (Abe, K., et. al. (1984) J. τιmmιτ.<ι_ . 132:1951) also showed inhibition, whereas MAbs directed to A antigen types other than the type 1 chain did not. Human lymphocytes are known not to produce endogenous blood group A and B antigens; however, these antigens may be acquired in the form of glycosphingolipids from plasma (Oriol, R., et. al. (1980) Hum. Tmmunoi. 1:195) or be expressed as a result of HIV induced neoglycosylation. The finding that monoclonal antibodies TKH2 and B72.3, directed to the sialyl-Tn antigen, inhibited infection and syncytium formation, suggests that O-linked glycans may also exist on the HIV envelope, as has been found in other enveloped viruses (Olofsson, S., et. al. (1981) Arch. Virol. 70:321). As no reduction in infection was found by preincubating target cells (MT4) prior to exposure to virus, the inhibitory effect of the MAbs seemed to be a result of binding to HIV rather than to the target cells. However, since TKH2 also labeled the target cells, the possibility that this antibody interfered with HIV infection by binding to the T4 receptor cannot be completely excluded. On the other hand, this is unlikely because monoclonal antibody B72.3, with similar specificity, failed to stain cells, but inhibited infectivity. The molecular nature of the glycoconjugates of the viral envelope interacting with the anti-carbohydrate monoclonal antibodies is at present obscure, and this interaction may involve other structures than gpl20. Most of the monoclonal antibodies which inhibited HIV did not label infected cells. This indicates that their epitopes were hidden until initial binding of gpl20 to the CD4 receptor has taken place, similar to what has been suggested for neutralizing anti-gp41 monoclonal antibodies (Kinney Thomas, E., et. al. (1988) AIDS 2:25). HIV-neutralizing antibodies are found in HIV-infected patients and have also been produced in vitro using selected synthetic peptides from gpl20 and gp41 (Weber J.N., et. al. (1989) Lancet i:ll9; Chanh, T.C, et. al. (1986) EMBO J. 5:3065; Lasky, L.A., et. al. (1986) Science 233:209). These do not seem to protect against disease progression and are type-specific. "Escape" mutants of HIV, containing mutations in the env gene, have been shown to arise in vitro, whereby the virus is able to escape the effect of formerly neutralizing antibodies (Weiss, R.A. (1988) J. Acq. Imm. Def. Syn. 1:536) . The anti-HIV reactivity of antisera against Equine infectious anemia virus is described against a carbohydrate part of the HIV envelope (Montelaro, R.C, et. al. (1988) J. Gen. Virol. 69:1711), but HIV-neutralization by carbohydrate-specific antibodies has not been described earlier. Carbohydrate epitopes may not be expected to be influenced as readily by mutations in the genome coding for the peptide part of the HIV envelope. Thus, the present results indicate that viral glycans can be considered targets for anti-viral immunotherapy and/or vaccine development. EXAMPLE 4 Production and Characterization of Monoclonal Antibodies to Sialyl-Tn Antigen Isolation of the immunogen Ovine submaxillary mucin (OSM) was used as the source of the sialyl-Tn antigen. Approximately 90% of the carbohydrate chain on OSM consists of the sialyl- Tn antigen. OSM was isolated from ovine submaxillary glands by conventional methods. (Tetta anti, G. and Pigman, W. (1968) Arch. Biochem. Biophys.. 124:45- 50). Briefly, aqueous extract of submaxillary glands was precipitated at acidic pH (e.g., 3.5). This is called a mucin clot. The mucin clot was centrifuged, dissolved in water, the pH adjusted to neutral, and fractional ethanol precipitation in sodium acetate was performed. The thus isolated OSM was used as was for the immunogen composition. Immunization and establishment of monoclonal antibodies The high molecular weight sialyl-Tn antigen isolated as described above was dissolved in distilled water in an amount of 4.0 mg protein/4 ml water and 16 mg acid-treated salmonella minnesota. obtained by conventional methods, was added. The mixture was thoroughly mixed for 1 hour at 57 β C, and then lyophilized. The lyophilized sample was then suspended in 4.0 ml of PBS and aliquots of 100 μl (i.e., 100 μg of the glycoprotein and 400 μg of bacteria) were injected intravenously through the caudal vein into each of five Balb/c mice. Injection was made once a week for five weeks and after one month of intermission, the mice were boosted with 200 μl of the antigen suspension (i.e., 200 μg of glycoprotein and 800 μg of the bacteria) . Three days after the booster injection, the animals were sacrificed, the spleen cells were removed and splenocytes were fused with mouse myeloma SP-2 cells by conventional methods (Hirohashi, S. et. al., (1984) Proc. Natl. Acad. Sci. USA. 82:7039-7043 and Fukushi, Y. et. al. (1984) J. Biol. Chem.. 259:4681- 4685) . Hybridomas which grew on selective media were screened by conventional methods for monoclonal antibody reactivity with the above-described OSM, -desialyated OSM, bovine submaxillary mucin (BSM) (approximately 50% of the carbohydrate chains on BSM consist of sialyl-Tn antigen) , and glycophorin A. Bovine submaxillary mucin (BSM) and glycophorin A were purchased from Sigma Chemical Company, St. Louis, Missouri. OSM was desialyated by treatment with 0.1 unit/ml of neurominodase from Clostridium Perfrinoens Type X (Sigma) by conventional methods. (Magnani, J.L. et. al. (1982) J. Biol. Chem.. 257:14365 and Fukushi et. al., (1984) J. Biol. Chem.. 259:10511) . Approximately 22 hybridomas were found which secreted monoclonal antibody that gave a positive reaction with OSM. These hybridomas were scaled up and the reactivity of the monoclonal antibodies was reexamined with OSM, desialylated OSM, BSM, and glycophorin A. Hybridomas secreting monoclonal antibodies showing strong reactivity with OSM, weak reactivity with BSM, and no reactivity with glycophorin A or sialidase-treated OSM were found. It should be understood that although the present invention has been described with a number of different embodiments, further modifications to the invention may be made. 47 Statement of Deposit The hybridoma cell lines BMl, AH21 and TKH2 (BM4) , which produce monoclonal antibodies BMl, AH21 and TKH2, respectively, have been deposited with the American Type Culture Collection, 12301 Parklawn Drive, Rockville, Maryland 20852 USA on December 19, 1989, September 14, 1989 and March 2, 1988, respectively and have ATCC deposit nos. HB 10312, HB 10226 and HB 9654, respectively. The deposits were made under the terms of the Budapest Treaty on the International Recognition of the Deposit of Microorganisms for the Purposes of Patent Procedure. All restrictions to access will be irrevocably removed upon grant of a U.S. patent on the instant application. IflttfMKwwl Λμ≠tcm*. H : PCT. 1-tttfMtoM. k ≠tMm Mβ: PCT/ InttrMtoMl Apf eMtM M#: PCT/

Claims: What is claimed is; l. A medicament for inhibiting or slowing the progression of acquired immune deficiency syndrome (AIDS) and AIDS related complex (ARC) comprising: (A) A pharmaceutically effective amount of one or more antibodies selected from the group consisting of anti-Le y , anti-A j and anti-sialyl-Tn, and (B) A pharmaceutically acceptable carrier, diluent or excipient. 2. The medicament of Claim 1, wherein said one or more antibodies comprises anti-Le y . 3. The medicament of Claim 2 , wherein said anti-Le y has an affinity for Le y corresponding to a KD of about 5x10 "7 . 4. The medicament of Claim 2, wherein said anti- Le y comprises one or more antibodies having the identifying characteristics of monoclonal antibody BMl produced by hybridoma BMl having ATCC deposit no. HB 10312. 5. The medicament of Claim 4/ wherein said anti-Le y comprises monoclonal antibody BMl produced by hybridoma BMl having ATCC deposit no. HB 10312. 6. The medicament of Claim 1 , wherein said one or more antibodies comprises anti-A-. 7. The medicament of Claim 6, wherein said anti-A- has the following identifying characteristics: (1) has an affinity for A j corresponding to a KD of about 10 "8 to 5xl0 "7 ; and (2) binds to at least type 1 chain A. 8. The medicament of Claim 7, wherein said anti-A j comprises one or more monoclonal antibodies having the identifying characteristics of monoclonal antibody AH21 produced by hybridoma AH21 having ATCC deposit no. HB 10226. 9. The medicament of Claim 8, wherein said anti-A j comprises AH21 produced by hybridoma AH21 having ATCC deposit no. HB 10226. 10. The medicament of Claim 7, wherein said anti-A j comprises one or more monoclonal antibodies having the identifying characteristics of monoclonal antibody AH16. 11. The medicament of Claim 10, wherein said anti-A- comprises monoclonal antibody AH16. 12. The medicament of Claim 1, wherein said one or more antibodies comprises anti-sialyl-Tn. 13. The medicament of Claim 12, wherein said anti-sialyl-Tn comprises one or more monoclonal antibodies having the identifying characteristics of monoclonal antibody TKH2 produced by hybridoma TKH2 having ATCC deposit no. HB 9654. 14. The medicament of Claim 13, wherein said anti-sialyl-Tn comprises monoclonal antibody TKH2 produced by hybridoma TKH2 having ATCC deposit no. HB 9654. 15. The medicament of Claim 12, wherein said anti-sialyl-Tn comprises one or more monoclonal antibodies having the identifying characteristics of monoclonal antibody TKH1. 16. The medicament of Claim 15, wherein said anti-sialyl-Tn comprises monoclonal antibody TKH1. 17. The medicament of Claim 12, wherein said anti-sialyl-Tn comprises one or more monoclonal antibodies having the identifying characteristics of monoclonal antibody B72.3. 18. The medicament of Claim 17, wherein said anti-sialyl-Tn comprises monoclonal antibody B72.3. 19. A method for inhibiting or slowing the progression of AIDS and ARC comprising administering to a subject having AIDS or ARC a medicament comprising: (A) A pharmaceutically effective amount of one or more antibodies selected from the group consisting of anti-Le y , anti-A j and anti-sialyl-Tn, and (B) A pharmaceutically acceptable carrier, diluent or excipient. 20. The method of Claim 19, wherein said one or more antibodies comprises anti-Le y . 21. The method of Claim 20, wherein said anti-Le y has an affinity for Le y corresponding to a KD of about 5xl0 "7 . 22. The method of Claim 20, wherein said anti- Le y comprises one or more antibodies having the identifying characteristics of monoclonal antibody BMl produced by hybridoma BMl having ATCC deposit no. HB 10312. 23. The method of Claim 22, wherein said anti- Le y comprises monoclonal antibody BMl produced by hybridoma BMl having ATCC deposit no. HB 10312. 24. The method of Claim 18, wherein said one or more antibodies comprises anti-A-. 25. The method of Claim 24, wherein said anti-A j has the following identifying characteristics: (1) has an affinity for A j corresponding to a KD of about 10 "8 to 5xl0 "7 ; and (2) binds to at least type 1 chain A. 26. The method of Claim 24, wherein said anti- A j comprises one or more monoclonal antibodies having the identifying characteristics of monoclonal antibody AH21 produced by hybridoma AH21 having ATCC deposit no. HB 10226. 27. The method of Claim 26, wherein said anti- A j comprises monoclonal antibody AH21 produced by hybridoma AH21 having ATCC deposit no. HB 10226. 28. The method of Claim 23, wherein said anti- A j comprises one or more monoclonal antibodies having the identifying characteristics of monoclonal antibody AH16. 29. The method of Claim 26, wherein said anti- A j comprises monoclonal antibody AH16. 30. The method of Claim 19, wherein said one or more antibodies comprises anti-sialyl-Tn. 31. The method of Claim 30, wherein said anti- sialyl-Tn comprises one or more monoclonal antibodies having the identifying characteristics of monoclonal antibody TKH2 produced by hybridoma TKH2 having ATCC deposit no. HB 9654. 32. The method of Claim 31, wherein said anti- sialyl-Tn comprises monoclonal antibody TKH2 produced by hybridoma TKH2 having ATCC deposit no. HB 9654. 33. The method of Claim 30, wherein said anti- sialyl-Tn comprises one or more monoclonal antibodies having the identifying characteristics of monoclonal antibody TKH1. 34. The method of Claim 33, wherein said anti- sialyl-Tn comprises monoclonal antibody TKH1. 35. The method of Claim 30, wherein said anti- sialyl-Tn comprises one or more monoclonal antibodies having the identifying characteristics of monoclonal antibody B72.3. 36. The method of Claim 35, wherein said anti- sialyl-Tn comprises monoclonal antibody B72.3. 37. A vaccine against AIDS and ARC, comprising: (A) An immunologically effective amount of one or more antigens selected from the group consisting of Le y , A j and sialyl-Tn, and (B) A pharmaceutically acceptable carrier, diluent or excipient. 38. The vaccine of Claim 37, wherein said one or more antigens comprises Le y . 39. The vaccine of Claim 37, wherein said one or more antigens comprises A.. 40. The vaccine of Claim 37, wherein said one or more antigens comprises sialyl-Tn. 41. A method of actively immunizing against AIDS and ARC, comprising administering to a subject a medicament comprising: (A) An immunologically effective amount of one or more antigens selected from the group consisting of Le y , A j and sialyl-Tn, and (B) A pharmaceutically acceptable carrier, diluent or excipient. 42. The method of Claim 41, wherein said one or more antigens comprises Le y . 43. A method of Claim 41, wherein said one or more antigens comprises A j . 44. The method of Claim 41, wherein said one or more antigens comprises sialyl-Tn.

Inventors: CLAUSEN HENRIK, HAKOMORI SEN-ITIROH, HANSEN JOHN-ERIK STIG, NIELSEN JENS OLE, VESTERGAARD BENT FABER, CLAUSEN, HENRIK, HAKOMORI, SEN-ITIROH, HANSEN, JOHN-ERIK, STIG, NIELSEN, JENS, OLE, VESTERGAARD, BENT, FABER
Applicants: BIOMEMBRANE INST, HANSEN JOHN ERIK STIG, NIELSEN JENS OLE, VESTERGAARD BENT FABER, HANSEN, JOHN-ERIK, STIG, NIELSEN, JENS, OLE, THE BIOMEMBRANE INSTITUTE, VESTERGAARD, BENT, FABER
Application year: 1991
Main classification: A61K39
IPC codes: A61K38, A61K39, A61P31, C07K14, C07K16, C12N5, C12N15, C12P21, C12R1