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Analysis of the Diversity of Murine Antibodies to Dextran B1355

I. Generation of a Large, Pauci-Clonal Response by a Bacterial Vaccine¹

Department of Microbiology and Immunology, and Department of Pathology, Washington University School of Medicine, St. Louis, Missouri 63110

Abstract

Mice immunized with a combination of dextran B1355 in adjuvant followed by three injections of 2 x 10⁹ Escherichia coli B organisms produced an average of 14.5 mg/ml of anti-dextran antibodies. It was demonstrated that the stimulating effect of E. coli B was due to antigenic determinants cross-reactive with B1355 and not solely because of adjuvant properties of the organism. The anti-dextran antibodies were distributed among both 7S and 19S components. Isoelectric focusing of the 7S antibodies showed several spectrotypes of antibody, most of which were shared by the majority of the individual sera. The limited spectrotypic heterogeneity of the 7S antibodies was supported by idiotypic studies. Thus, a heterologous, anti-idiotypic serum, rabbit anti-M104, was prepared which distinguished between two closely related myeloma proteins, M104 and J558, with specificity for -(1 3) dextran. This antiserum demonstrated that some, but not all, of the 7S and 19S anti-dextran antibodies possessed variable region determinants cross-reactive with M104.

Footnotes

¹ This work was supported by United States Public Health Service Grants AI-11635 and CA-16032 and by the following companies: Brown and Williamson Tobacco Corporation; Larus and Brother Company, Inc.; Liggett and Myers, Inc.; Lorillard, a Division of Loews Theatres, Inc.; Philip Morris, Inc.; R. J. Reynolds Tobacco Co.; United States Tobacco Co.; and Tobacco Associates, Inc.

² Recipient of United States Public Health Service Training Grant GM-00897.

³ Recipient of United States Public Health Service Postdoctoral Fellowship AI-00685.