Cultured human T lymphocyte lines and clones as immunogenetic tools

Human T lymphocytes were sensitized in vitro to allogeneic determinants encoded by the HLA region and subsequently cloned by limiting dilution in the presence of human T cell growth factor. An in vitro priming protocol entailing multiple rounds of allostimulation in mixed lymphocyte culture, using a disparate stimulating to responding cell ratio (10:1), enabled a high frequency of functionally active clones to be isolated. The specificity of the proliferative and cytotoxic reactivities was ascertained directly by screening in an HLA characterized family.