Colorimetric B cell proliferation assay based on alkaline phosphatase activity. Selective measurement of B cell proliferation in the presence of other cell types.
In this study we show that a rapid colorimetric determination of alkaline phosphatase (APase) activity can be used in conjunction with a multiwell scanning photometer to quantitatively measure lymphokine-dependent B cell proliferation. Despite the fact that only activated but not resting B cells exhibit APase activity, i.e., that APase expression in B cells is variable, the results obtained by monitoring the enzyme activity were very similar to those obtained by measuring thymidine incorporation in two murine B cell systems. With regard to a potential application to the screening of the effects of hybridoma supernatants on B cells, it is relevant that HAT medium has practically no influence on the colorimetric B cell proliferation assay. Another advantage of the method is that since T cells and macrophages lack APase activity, one can detect the proliferation of B cells even in the presence of a high number of such cells, for example, in studying a T cell-dependent response.
D000469:Alkaline Phosphatase / Q000032:analysis*; D000818:Animals; D001402:B-Lymphocytes / Q000166:cytology* / Q000187:drug effects / Q000201:enzymology; D002478:Cells, Cultured; D003124:Colorimetry / Q000379:methods*; D004261:DNA Replication; D007041:Hypoxanthine Phosphoribosyltransferase / Q000378:metabolism; D008213:Lymphocyte Activation / Q000187:drug effects; D008222:Lymphokines / Q000494:pharmacology; D008565:Membrane Proteins / Q000032:analysis; D051379:Mice; D008810:Mice, Inbred C57BL; D013601:T-Lymphocytes; D013936:Thymidine / Q000378:metabolism
D008222:Lymphokines; D008565:Membrane Proteins; D007041:Hypoxanthine Phosphoribosyltransferase; D000469:Alkaline Phosphatase; D013936:Thymidine