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An IgG₃ Monoclonal Antibody Established after Immunization with G_M3 Lactone: Immunochemical Specificity and Inhibition of Melanoma Cell Growth in Vitro and in Vivo¹

The Biomembrane Institute; Department of Biochemical Oncology, Fred Hutchinson Cancer Research Center; and Departments of Pathobiology and Microbiology, University of Washington, Seattle, Washington 98119

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 G_M3 (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 G_M3 lactone (Nores, G. A., et al., J. Immunol., 139: 3171–3176, 1987). Since G_M3 lactone was found to be highly immunogenic, we subsequently immunized C57BL/6 mice with G_M3 lactone coated on Salmonella minnesotae and established hybridoma DH2, secreting an IgG₃ antibody showing preferential reactivity with G_M3 lactone over G_M3 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).

¹ This investigation has been supported by an Outstanding Investigator Grant, CA42505 from the National Cancer Institute, and funds from The Biomembrane Institute.

² Recipient of a fellowship from Consejo Nacional de Investigaciones Cientificas y Técnicas de la República Argentina (1985–1986).

³ To whom requests for reprints should be addressed, at The Biomembrane Institute, 201 Elliott Avenue West, Seattle, WA 98119.

Received 3/25/88. Revised 7/15/88. Accepted 7/19/88.

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