Quantitative analyses of specific B cells for sheep red blood cell, phosphorylcholine, and hepatitis B surface antigen in human B cell populations by polyclonal transformation with Epstein-Barr virus.

Abstract

An experimental system was developed to analyze quantitatively specific B cells for various antigens in human B cell populations by polyclonal transformation and Ig production induced by in vitro infection with Epstein-Barr virus (EBV). EBV-infected cells were cultured in microtest plates for 3 wk and specific antibody activities for sheep red blood cell (SRBC) were detected in the culture supernatants by hemolysis and those for phosphorylcholine (PC) and for hepatitis B surface antigen (HBsAg) by passive hemagglutination. Frequencies of positive cultures with specific antibodies followed one hit-type of Poisson''s distribution. Large numbers of cultures showed specific antibody activities for SRBC and for PC with tonsillar lymphocytes obtained from 4 donors. Positive cultures with anti-HBsAg antibodies were practically undetectable with 3 donors. With the 4th donor, a large fraction of cultures showed specific antibody activities for HBsAg. Frequencies of positive cultures with anti-HBsAg antibodies were further analyzed with peripheral blood lymphocytes obtained from 5 donors with serum anti-HBsAg. Comparable frequencies of cultures were positive with anti-SRBC antibodies with these donors. The frequencies of cultures with anti-HBsAg antibodies were quite variable among these donors. These results directly reflected the sizes of clones with these specificities in human lymphocyte populations obtained from donors with various immunological histories.