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Studies on the Terminal Stages of Immune Hemolysis

V. Evidence That Not All Complement-Produced Transmembrane Channels Are Equal

From the Laboratory of Immunobiology, National Cancer Institute, Bethesda, Maryland 20014

Abstract

The inhibitory effects of 0.1 M EDTA on the lysis of E* prepared by incubating EA with whole GPC was studied. At high end point lysis (>70%) 0.1 M EDTA failed to prevent hemoglobin release whereas at lower end point (<60%) 0.1 M EDTA was effective. In all cases hemoglobin release was inhibited by 25% BSA. When E* were prepared by incubating EAC1-8 with C9, similar results were obtained. In this system the difference in the ability of 0.1 M EDTA to inhibit hemoglobin release at high or low end point lysis could not be correlated with the number of lesions/cell but appeared to be related to the C9 to SAC1-8 ratio. With limiting SAC1-8 and excess C9, E* were produced from which hemoglobin release could not be prevented by 0.1 M EDTA whereas at lower C9 to SAC1-8 ratios hemoglobin release was prevented by 0.1 M EDTA. These differences most probably reflect functionally different sized transmembrane channels that were produced at different C9 to SAC1-8 ratios.