Isolation of HLA-haplotype-specific human T-lymphocyte clones

Human T-lymphocyte clones showing proliferative and cytotoxic responses specific for HLA antigens were isolated. To obtain enriched numbers of functionally active colonies, the cells were first sensitized in vitro in a tertiary mixed lymphocyte culture (MLC), using 10 stimulating cells for each responding cell. To assess quickly the genetic specificity of the clones, the cells used in the priming MLC and later as screening cells were prepared from an HLA-typed family. Using this approach, 75% of the colonies seeded at 1 cell/well showed functional activity specific for HLA.