Biochemical and functional characterization of mature progeny purified from a myelomonocytic leukemia cell line.
Comparative studies were performed on the cloned myelomonocytic leukemia cell line, WEHI-3B, and a subcloned line, WEHI-3BM6. WEHI-3BM6 cells were less responsive than WEHI-3B cells to differentiation factor (DF) present in the sera of mice injected with endotoxin (endotoxin serum, ES). WEHI-3BM6 cells produced only 8% monocytes after 6 days of incubation with ES compared with 68% monocytes produced by WEHI-3B cells. In the presence of ES the rate of differentiation of both cell lines was enhanced by the addition of actinomycin D (5 ng/ml) such that after 2 days of stimulation 62% of the cells were mature monocytes. Lysozyme content as well as the expression of alpha-napthyl acetate esterase were also increased by actinomycin D. As indicated by the shifts in modal fluoresence levels (209 vs 63), differentiating WEHI-3B cells showed an increase in the binding of an anti-neutrophil serum compared with untreated WEHI-3B cells. The binding of anti-neutrophil antibodies allowed the sorting of the mature monocytes from the blast cells in DF-treated WEHI-3B cells achieving a purity of 78%. Electrophoretic analysis of radiolabelled proteins from the cell extracts of WEHI-3B-derived monocytes showed close similarities to normal murine peritoneal macrophages and distinct differences from the protein profiles of purified murine peritoneal polymorphs. A protein of 75,000 mol, wt present in the polymorphs was absent from the WEHI-3B monocytes.
D000818:Animals; D002452:Cell Count; D002454:Cell Differentiation / Q000187:drug effects; D002455:Cell Division; D002460:Cell Line; D002469:Cell Separation; D002999:Clone Cells; D003609:Dactinomycin / Q000494:pharmacology; D004950:Esterases / Q000032:analysis; D007106:Immune Sera; D007951:Leukemia, Myeloid / Q000473:pathology*; D051379:Mice; D008807:Mice, Inbred BALB C; D009113:Muramidase / Q000032:analysis
D007106:Immune Sera; D003609:Dactinomycin; D004950:Esterases; D009113:Muramidase