Fractionation of Antigen Reactive Cells on a Cellular Immunoadsorbent: Factors Determining Recognition of Antigens by T-Lymphocytes

作者：Hartmut Wekerle, Peter Lonai, M. Feldman · 发表于：Proceedings of the National Academy of Sciences · 年份：1972 · DOI：10.1073/pnas.69.6.1620 · 被引用次数：56 · 研究领域：Monoclonal and Polyclonal Antibodies Research、RNA Interference and Gene Delivery、Immunotherapy and Immune Responses

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 .