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Characterization of a glycosyl-phosphatidylinositol anchor-deficient subline of Raji cells. An analysis of the functional importance of complement inhibitors on the Raji cell line.

Authors
  • C L Harris
  • B P Morgan
Publication Date
Oct 01, 1995
Source
PMC
Keywords
Disciplines
  • Biology
License
Unknown

Abstract

Analysis of complement inhibitory proteins present on the surface of Raji cells (obtained from the European Collection of Animal Cell Cultures; originally established from human Burkitt's lymphoma) revealed two populations of cells. These populations differed in their expression of the glycosyl-phosphatidylinositol (GPI)-anchored inhibitors CD59 and decay-accelerating factor (DAF). Two stable clones were established by limiting dilution of the original cell culture. Raji+3 expressed CD59 and DAF whereas Raji-26 expressed neither inhibitor. Both clones expressed membrane cofactor protein (MCP). Analyses of other cell surface proteins (CD19, CD35, CD48 and CD58 (transmembrane form)) revealed similar levels of expression of transmembrane proteins by both clones. However, CD48 was expressed only by Raji+3. As CD48, DAF and CD59 are all GPI-anchored molecules it is likely that a defect in the GPI-anchoring mechanism is responsible for the generation of the second population of cells. The two clones demonstrated markedly different sensitivities to complement. When equally sensitized cells from both clones were treated with normal human serum (12.5%) for 1 hr at 37 degrees, the Raji+3 clone was resistant to complement-mediated lysis, whereas approximately 70% of the Raji-26 cells were lysed. However, by using specific antibody to block the function of membrane-bound complement inhibitors, lysis of Raji+3 was demonstrated. Whilst blocking of one inhibitor only on the cell had little effect on cell killing, blocking of two or more inhibitors significantly increased cell lysis. Our results demonstrated that all three inhibitors expressed by these cells contributed to protection against classical pathway-mediated complement activation. However, whilst a limited protective role was seen for MCP, CD59 and DAF appeared to be of far more importance for protection from complement-mediated lysis via the classical pathway.

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