Differentiation of chronic lymphocytic leukemia cells after in vitro treatment with Epstein-Barr virus or phorbol...

Leukemic cells from ten patients with "nonsecretory," B-type chronic lymphocytic leukemia (CLL) were cultured alone or in the presence of Epstein-Barr virus (EBV) or a phorbol ester (12-0-tetradecanoylphorbol-13-acetate; TPA) for 7 days. At periodic intervals the cell morphology, cytoplasmic immunoglobulin content (direct immunofluorescence) and capacity for immunoglobulin secretion (hemolytic plaque assay) were assessed. A variable but significant number of the TPA-treated CLL cells from all patients expressed cytoplasmic immunoglobulin of a single light-chain type at some stage, usually within the first 3 days. EBV induced similar changes in seven of eight cases tested. Untreated cell cultures were negative or contained a few cytoplasmic immunoglobulin-positive cells. Cells from five of nine cases secreted immunoglobulin of a single light-chain type. In every instance this was identical to the surface and cytoplasmic immunoglobulin. Cytologic changes were observed in the leukemic cells after treatment with one or both agents in nine of ten cases. The major feature was an increase in cell size associated with immunoblastic or plasma-cytoid features. Mitotic figures and binucleate cells were also present. These studies indicate that EBV and TPA are effective at inducing immunoglobulin synthesis and secretion in "nonsecretory" B cell neoplasms and are useful tools for studying the maturation potential inherent in these tumors. The study also suggests that the secreted immunoglobulin is a monoclonal product.