Glycosylphosphatidylinositol (GPI)-deficient Jurkat T cells as a model to study functions of GPI-anchored proteins.
Many cell surface proteins attached to the membrane by GPI are involved in cell signalling. However, the role of the GPI membrane anchor itself remains poorly understood. GPI-defective cells from patients with paroxysmal nocturnal haemoglobinuria (PNH) are relatively resistant to apoptosis induction. We developed a Jurkat T cell model for GPI deficiency by isolating a GPI-negative mutant, which is defective in the GPI biosynthetic gene PIG-A. Using retroviral PIG-A gene transfer along with the transfer of a vector control, we obtained two genetically identical cell lines, distinguished only by expression of the PIG-A gene and, thus, their ability to produce GPI. Cell proliferation and survival were not affected by this difference. Apoptotic stimuli such as serum starvation and camptothecin exposure elicited similar responses. In contrast, GPI-defective Jurkat cells were more susceptible to Fas-mediated apoptosis than GPI-positive cells. These results indicate that a deficiency in GPI-anchored proteins, as is found in PNH, does not confer resistance to apoptosis.
Bastisch
I
I
Department of Clinical Immunology, Hannover Medical School, Hannover, Germany.
Tiede
A
A
Deckert
M
M
Ziolek
A
A
Schmidt
R E
RE
Schubert
J
J
D017209:Apoptosis / Q000187:drug effects; D002166:Camptothecin / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005822:Genetic Vectors; D017261:Glycosylphosphatidylinositols / Q000096:biosynthesis / Q000378:metabolism*; D006801:Humans; D019169:Jurkat Cells; D008565:Membrane Proteins / Q000235:genetics / Q000502:physiology*; D008954:Models, Biological; D012190:Retroviridae; D059004:Topoisomerase I Inhibitors; D019014:fas Receptor / Q000276:immunology / Q000378:metabolism
D004791:Enzyme Inhibitors; D017261:Glycosylphosphatidylinositols; D008565:Membrane Proteins; D059004:Topoisomerase I Inhibitors; D019014:fas Receptor; C080086:phosphatidylinositol glycan-class A protein; D002166:Camptothecin