Fractionation of antigen reactive cells on a cellular immunoadsorbent: factors determining recognition of antigens by T-lymphocytes.

Monolayers of mouse fibroblasts were used as cellular immunoadsorbents to separate rat lymphocytes that recognize specific mouse histocompatibility antigens. Normal lymphocytes were incubated on fibroblasts of strain C3H/eb, and nonadherent cells were separated from adherent cells, then transferred for sensitization onto fresh monolayers of C3H. When tested on (51)Cr-labeled target monolayers the nonadherent cells manifested significantly lower cytotoxicity than the adherent cells. However, the nonadherent cells could be sensitized against mouse fibroblasts of an unrelated H-2 phenotype (strain Balb/c). The immune specificity of the adherence was further demonstrated by a stepwise adsorption, which resulted in complete loss of the capacity to acquire specific lytic activity towards C3H antigens. Lymphocytes recognizing strain-specific antigens of the mouse were separated at 37 degrees (not at 0 degrees or 4 degrees ); separation was inhibited after treatment with dinitrophenol, sodium azide, or trypsin. Prior treatment with neuraminidase rendered the lymphocytes suceptible to separation at 0 degrees .