Abstract
We previously reported that HSV can induce T-dependent in vitro immunoglobulin secretion in individuals sero “+” to HSV (Immunobiology 162:94, 1982). This study shows that 12-day culture supernatants from peripheral blood mononuclear cells or T and B cells cultured in RPMI 1640 with antibiotics and fetal calf serum stimulated with HSV produce IgG anti-HSV which can be detected using an ELISA (detects high titer anti-HSV serum at a dilution of 10−12). Since an individual's immune status, precursor frequency of antigen specific T and B cells, antigen dose, and culture conditions can limit antigen-specific ab responses, culture supernatants were screened for specific ab from 15 sero “+” (titer ≥ 10−5 by ELISA) and 7 “-” individuals at varying cell concentrations, at varying doses of antigen and from different types of culture vessels. Our results show that net ab synthesis occurred: in lymphocytes from 10 of 15 sero “+” and none of 7 saro “-” individuals; at cell concentrations between 0.5-0.8 x 106 cells/well; in round bottom microtiter wells; at T:B ratios from 0.5:1 to 3:1; and optimally after washout of HSV after 4 days of culture. Replicate cultures were needed for detecting ab. These findings illustrate the immunobiology of in vitro specific ab synthesis in that high numbers of in vivo sensitized antigen-specific precursors T and B cells cultured in high density configurations after antigen washout to prevent suppression of ab synthesis were necessary for ab synthesis.
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