Polyclonal activation of bone-marrow-derived lymphocytes from human peripheral blood measured by a direct plaque-forming cell assay.

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Abstract
A culture and assay system for the stimulation of human peripheral blood lymphocytes with polyclonal activators of bone-marrow-derived (B) lymphocytes (e.g., pokeweed mitogen and Escherichia coli lipopolysaccharide) and subsequent measurement of single cell antibody production by a hemolysis-in-gel direct plaque-forming cell assay against sheep erythrocytes was established. The critical culture requirement were delineated and a new highly sensitive ultrathin gel assay method was described. Under these conditions, a substantial and highly reproducible plaque-forming cell response was detected in normal human peripheral blood. This system can be readily used to explore the complex events associated with activation of human B cells.