High frequency detection of different T-cell subsets in mice by a modified virus plaque assay

Different T-cell subsets participating in immune responses were detected at a high frequency by a modified virus plaque assay (VPA). By using the modified VPA, different activated T-cell subsets generated in primary immune responses, helper and suppressor T cells participating in antibody formation, and effector T cells involved in the delayed-type hypersensitivity (DTH) reaction were enumerated directly without in vitro antigen stimulation. The frequency of detection in immune systems used was 7.5-17.7 V-PFC/10(3) spleen cells. Although neither helper T cells for antibody formation nor effector T cells for DTH reaction were detected as V-PFC at a high frequency by the original VPA, it was also found in secondary immune response that Lyt 1 positive, antigen-specific helper and effector T-cell subsets, and cyclophosphamide (CY)-resistant precursors were enumerated at a high frequency by the modified VPA when the received in vitro antigen stimulation, and that the proliferative stage of these cells was critical for the development of V-PFC.