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Complement: Chemical Coupling of a Functionally Active Component to Erythrocytes in the Absence of Antibody¹

From the Department of Microbiology and the Pediatric Immunology Unit of the Section of Hematology and Immunology, Department of Medicine, University of California, San Francisco Medical Center, San Francisco, California 94122

Abstract

Unsensitized sheep erythrocytes exposed to the first component of complement (C'1) and tannic acid, bis-diazotized benzidine (BDB), or CrCl₃ were lysed by whole guinea pig C', but similarly treated cells not exposed to C'1 were not lysed. Lysis of the tanned, C'1-treated cells was blocked by: a) heat-inactivating the C'1; b) diluting the C' in EDTA; c) washing the cells with EDTA after exposure to C'1 but before exposure to whole C'; and d) exposing the cells to a high concentration of protein before adding the C'1. C'1 could be demonstrated on tanned C'1-treated cells by the C'1 transfer technique. The C'1 coupled to cells by BDB or CrCl₃ appeared to be more tightly bound than with tannic acid, as indicated by failure to transfer C'1 from these cells and by retention of cell-bound reactivity after washing with EDTA. BDB and CrCl₃, and to a lesser extent tannic acid, were quite destructive of fluid-phase C'1. Coupling of functionally active C'1 to red cell membranes may account for earlier observations on the lysis of tanned erythrocytes by C' in the absence of antibody.

Footnotes

This study was supported by grants from the National Institutes of Health (AI-06464-03S1, 5 T01 AI-00299 and AM-08527).

² Helen Hay Whitney Fellow.

³ Second-year medical student.