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Fixation of C' and C'la by Rabbit G- and M-Antibodies with Particulate and Soluble Antigens¹

From the Children's Asthma Research Institute and Hospital, Denver, Colorado

Abstract

C' and C'1a fixation by G- and M-antibodies was studied with A substance and type A human erythrocytes as antigens and specifically purified rabbit antibodies. In soluble antigen systems, the maximal C' fixation by G-antibody was obtained at the equivalence antigen to antibody ratio, whereas the maximal fixation by M-antibody was obtained in excess antibody. At the optimal antigen to antibody ratio for C' fixation, the C'-fixing activity of the G- and M-antibodies was comparable on a molar basis. Essentially no C' fixation was observed by soluble antigen-M-antibody complexes formed in excess antigen. Comparisons of C'1a fixation by G- and M-antibodies in both soluble and cell antigen systems indicated that M-antibody is more effective than G-antibody on the cell surface, while the effect was reversed when the soluble antigen was used. It was confirmed that a single M-antibody molecule on the cell surface established one C'1a-fixing site. The M-antibody complexes with insoluble A substance had much higher C' and C'1a-fixing activities than those combined with soluble A substance. The different efficiency of C'1a fixation by M-antibody, depending on the nature of antigen, was ascribed to the number and distribution of antigenic determinants on the surface of antigen involved. The results strongly suggested that a combination of M-antibody molecules with antigen through multiple combining sites is essential for the induction of C'-fixing activity by M-antibody.

Footnotes

This work was supported by Grant AI-04985 from the United States Public Health Service and presented before the 51st Annual Meeting of the American Assocation of Immunologists in Chicago, Ill., April 1967.

² Present address: Chiba University, The Medical School, Department of Pathology, Chiba-city, Japan.

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