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The Journal of Immunology, Vol 151, Issue 8 4295-4305, Copyright © 1993 by American Association of Immunologists

ARTICLES

Mouse Crry/p65. Characterization of monoclonal antibodies and the tissue distribution of a functional homologue of human MCP and DAF

B Li, C Sallee, M Dehoff, S Foley, H Molina and VM Holers
Howard Hughes Medical Institute, Washington University School of Medicine, St. Louis, MO 63110.

In previous studies we have demonstrated that mouse Crry/p65 regulates complement component C3 deposition on self membranes, a functional property that both human decay-accelerating factor (DAF) and membrane cofactor protein (MCP) exhibit. We have proposed that Crry/p65 has a similar biologic role in mouse as MCP and perhaps DAF and is the mouse analogue of one or both of these proteins. In order to address this hypothesis and further study Crry/p65, we have prepared rat mAb and a rabbit polyclonal Ab to this protein. Using these reagents we demonstrate that, like human MCP and DAF, the tissue distribution of Crry/p65 is very broad. Most if not all cells of nonneuronal origin express this protein. In addition, by immunohistochemical analysis, Crry/p65 is shown to be more highly expressed in some tissues at potential sites of immune complex deposition and damage, such as the mesangium of the renal glomerulus and the arterial vessel endothelium. By Western blot analysis, protein isoforms can be demonstrated. Unlike human DAF, however, no phosphatidylinositol-specific phospholipase C- sensitive Crry/p65 protein form can be demonstrated on lymphocytes or erythrocytes. Five of six anti-Crry/p65 mAb can partially or completely reverse the capacity of Crry/p65 to block C3 deposition on cell membranes. Analysis of four IgG rat anti-Crry/p65 mAb demonstrates that two major independent epitopes can be detected. Overall, Crry/p65 retains many of the major features of human MCP and DAF, and the use of these reagents should further the understanding of the biologic roles of this class of complement regulatory proteins.


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