Long-term maintenance of HLA-D restricted T cells specific for soluble antigens

Lymphocytes from donors sensitized to soluble protein antigens tuberculin (PPD) and tetanus toxoid were stimulated in vitro with these antigens. The blasts were isolated on density gradients and maintained in long-term proliferating culture by the addition of supernatants from phytohaemagglutinin-stimulated (PHA) cultures. Blasts can be shown to retain specificity for the original stimulating antigen as measured by stimulation of DNA synthesis, but only when the antigen is presented in the company of a cooperating cell population. Autologous irradiated peripheral blood lymphocytes provide the best cooperation, but donors who share HLA-D antigens will also allow for continued proliferation in the presence of the appropriate soluble antigen. Donors sharing at HLA-A, -B, or -C show minimal ability to cooperate. The soluble antigen-specific blast cells do not manifest alloreactivity. The data are discussed with regard to possible application to clinical histocompatibility typing and to the implications of selfrecognition in the immune response.