An IgG3 monoclonal antibody established after immunization with GM3 lactone: immunochemical specificity and inhibition of melanoma cell growth in vitro and in vivo

Abstract

In previous studies, an IgM monoclonal antibody (M2590), established after immunization of C57BL/6 mice with syngeneic B16 melanoma cells, was found to react with melanoma cells, but not with various normal cells and tissues (Taniguchi, M., and Wakabayashi, S., Jpn. J. Cancer Res., 75:418-426, 1984). The structure defined by this antibody was identified as GM3 (Hirabayashi, Y., et al., J. Biol. Chem., 260:13328-13333, 1985) organized in membranes at high density, although the real immunogen was suggested to be GM3 lactone (Nores, G. A., et al., J. Immunol., 139:3171-3176, 1987). Since GM3 lactone was found to be highly immunogenic, we subsequently immunized C57BL/6 mice with GM3 lactone coated on Salmonella minnesotae and established hybridoma DH2, secreting an IgG3 antibody showing preferential reactivity with GM3 lactone over GM3 under certain conditions. The reactivity of the DH2 antibody was competitively inhibited by M2590, and it showed a preferential reactivity with melanoma cells and displayed various immunochemical and immunobiological properties similar to those of M2590. However, DH2 antibody inhibited melanoma cell growth in vivo, induced antibody-dependent cytotoxicity in vitro, and showed a preferential accumulation in melanoma growth in vivo. These properties are characteristic of the IgG3 subclass, in striking contrast to IgM antibody M2590, which does not inhibit cell growth in vivo or in vitro and does not induce antibody-dependent cytotoxicity. Thus, immunization with lactone forms of tumor-associated ganglioside antigens might be useful in the production of antibodies and prevention of tumor cell growth in vivo (antitumor vaccines).