Complement alternative pathway activation and control on membranes of human lymphoid B cell lines.
Membrane regulatory molecules normally prevent complement activation by autologous cells, therefore we compared the membrane control system of human lymphoid cell lines which activate or not human complement through the alternative pathway (AP). Membrane expression of decay-accelerating factor (DAF), membrane cofactor protein (MCP), complement receptors (CR)1, CR2 and H was measured either by radioimmunoassay or enzyme-linked immunosorbent assay on cell lysates. Soluble extracts of isolated membranes were tested functionally for their ability to accelerate the decay of C3bBb C3-convertase and allow the cleavage of C3b by factor I. Both regulatory functions were detected in solubilized membranes of Ramos cells, which do not activate the AP, as well as on the potent AP activator, Raji. Raji cells were found to express CR2, DAF and MCP molecules, while MCP was the only known regulatory protein detected on Ramos cells which expressed neither CR1, nor CR2, H or DAF. The I-cofactor activity of both Raji and Ramos cells was immunoprecipitated by anti-MCP, but the decay-accelerating activity was not adsorbed by anti-DAF nor by any of the available antibodies. Two EBV genome-negative cell lines (BJAB, BL41) were tested before and after in vitro conversion by EBV. As previously shown, EBV-converted cell lines activate the AP more efficiently than EBV- cell lines. At the same time, EBV superinfection induces an increase of both AP regulatory functions of cell membranes and enhances the expression of DAF, MCP and CR2. The results of this study show that complement activation by lymphoid cell lines is not related to an impaired autologous control of these cells, but that the expression of regulatory molecules increases together with the appearance of activating structures on the cell surface. Our results also suggest the occurrence of a new factor involved in the decay-accelerating activity on BL lines.
Caudwell
V
V
INSERM U25, Hôpital Necker, Paris, France.
Porteu
F
F
Calender
A
A
Pangburn
M K
MK
Halbwachs-Mecarelli
L
L
D015703:Antigens, CD; D000944:Antigens, Differentiation, B-Lymphocyte / Q000502:physiology; D001402:B-Lymphocytes / Q000502:physiology*; D018958:CD55 Antigens; D002460:Cell Line; D002462:Cell Membrane / Q000502:physiology; D050577:Complement C3-C5 Convertases / Q000378:metabolism; D003179:Complement C3b / Q000378:metabolism; D003180:Complement C3b Inactivator Proteins / Q000502:physiology; D017242:Complement Factor H; D017244:Complement Factor I; D003170:Complement Pathway, Alternative; D004854:Herpesvirus 4, Human / Q000276:immunology; D006801:Humans; D066298:In Vitro Techniques; D051916:Membrane Cofactor Protein; D008562:Membrane Glycoproteins / Q000378:metabolism; D008565:Membrane Proteins / Q000502:physiology; D011951:Receptors, Complement / Q000502:physiology; D017463:Receptors, Complement 3b; D017464:Receptors, Complement 3d; D012697:Serine Endopeptidases / Q000378:metabolism; D014412:Tumor Virus Infections / Q000276:immunology
D015703:Antigens, CD; D000944:Antigens, Differentiation, B-Lymphocyte; C497857:CD46 protein, human; D018958:CD55 Antigens; C493495:CFH protein, human; D003180:Complement C3b Inactivator Proteins; D051916:Membrane Cofactor Protein; D008562:Membrane Glycoproteins; D008565:Membrane Proteins; D011951:Receptors, Complement; D017463:Receptors, Complement 3b; D017464:Receptors, Complement 3d; D003179:Complement C3b; D017242:Complement Factor H; D050577:Complement C3-C5 Convertases; D012697:Serine Endopeptidases; D017244:Complement Factor I