Isolation and characterisation of a CDw50 negative Jurkat T-cell line variant (PPL.1)

PPL.1, a Jurkat cell line variant deficient in CDw50 surface expression, has been selected by fluorescence-activated cell sorting and expanded in cell culture. We have studied the expression of several leukocyte surface markers (CD2, CD3, CD4, CD8, CD7, CD26, CD25, CD14, CD18, kCD20, CD43, CD45, CD45R, CD71 and HLA class I and II) and we find no differences in their expression between PPL.1 and its parental Jurkat cell line. Immunoprecipitation analysis of metabolically labelled PPL.1 cells ([35S]-cysteine plus [35S]-methionine) fails to detect the presence of a preformed cytoplasmic pool of CDw50 molecules. The deficient CDw50 expression on PPL.1 cells is stable after several weeks of continuous culture and even after exposure of cells to several lymphocyte activating agents (PGE2, PHA, Con A, calcium ionophore A23187 and human recombinant IFN-gamma). No karyotype changes responsible for such phenotype deficiency are found. PPL.1 cells are as efficient as wild-type Jurkat or K562 cells, when used as targets in cytotoxicity assays with fresh or PHA-stimulated peripheral blood lymphocytes. No blocking effects of CDw50-specific mAb are observed in such assay. These results are consistent with the fact that CDw50 is not involved in alloreactive T-cell-specific cytotoxicity. They also suggest that this antigen is implicated only on a very specialized type of cell-cell interactions.