A micro-method for the study of in vitro lymphocyte transformation to specific antigens

A quantitative method for studying blast-transformation of cultured lymphocytes in the presence of specific antigens is described. The technique is performed in 0.3 ml microtrays, using 2.5 X 10(5) lymphocytes per culture, with an optimal culture period of 6 days. Cultures are pulsed for 4 h with [3H]-thymidine, giving maximum incorporation and minimum cellular damage. The study also describes some disadvantages of using lymphocyte responses to phytohaemagglutinin (PHA), as a marker of T-cell function. Experiments to establish ideal conditions for each stage of the method are described. The technique can be adjusted to accommodate different antigens, cell types and for the study of animal as well as human lymphocytes.