INDUCTION OF PEPTIDE-SPECIFIC PRIMARY CYTOTOXIC T-LYMPHOCYTE RESPONSES FROM HUMAN PERIPHERAL-BLOOD

Various protocols were developed and compared for eliciting specific c ytotoxic T lymphocyte (CTL) cell lines from the unselected human perip heral blood mononuclear cells of naive donors. Interleukin-7 and CD4() T cells primed in vitro by keyhole limpet hemocyanin were shown to a ct together in the generation of these responses. Primary responses we re consistently induced with a variety of different HLA class I-bindin g malarial peptides. Primary CTL responses could be induced from unsel ected CD8(+) and from CD45RA(+)CD8(+) T cells. The CTL lines derived f rom these naive donors were CD8(+) and demonstrated a high level of HL A class I-restricted killing for > 3 months after priming in vitro. Th ey were also able to recognize and kill targets infected with a recomb inant vaccinia virus containing the full-length antigen. In addition, this same protocol enhanced up to fourfold the levels of secondary CTL responses induced. The optimal method presented for naive cytotoxic T cell stimulation is simple, rapid and generally applicable and should provide a useful tool for both basic research and human therapy.