Skip to main content
U.S. Department of Energy
Office of Scientific and Technical Information

Killing of human melanoma cells by the membrane attack complex (MAC) as a function of its molecular composition

Conference · · Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States)
OSTI ID:5495592

The efficiency of the MAC in causing membrane damage and lysis of M21 cells was determined varying the C9:C8 molar ratio. C8,C9-depleted serum was reconstituted with radio-labeled C8 and C9 in kinetic and dose-response analyses. C5b-8 was sufficient to cause membrane leakiness, determined by /sup 86/Rb release and propidium iodide (PI) uptake, and this membrane damage was largely reversible. Cell killing was effected at high C5b-8 density as evidenced by /sup 51/Cr release and PI uptake with enhancement by C9. C5b-8 dependent cytolysis showed a lag between C8 binding and functional channel formation and increased with C8 uptake, suggesting cooperativity of C5b-8 in channel formation. With limiting numbers of bound C8 molecules, C9 binding and channel formation proceeded without lag. Saturation of C9 binding sites was observed at an average C9:C8 ratio of 6:1. By electron microscopy (EM), circular membrane lesions were visualized at a C9:C8 ratio of 3:1 or higher. Maximal cytolytic efficiency of MAC was observed at lower ratios at which ultrastructural lesions could not be detected. These results suggest that M21 cells can be killed by C5b-8 and that killing is maximized by amounts of C9 insufficient for poly C9 formation as detected by EM.

Research Organization:
Research Institute of Scripps Clinic and UCSD Medical Center, La Jolla, CA
OSTI ID:
5495592
Report Number(s):
CONF-8604222-
Journal Information:
Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States), Journal Name: Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States) Vol. 45:3; ISSN FEPRA
Country of Publication:
United States
Language:
English