In vitro methods for generating CD8+ T-cell clones for immunotherapy from the naïve repertoire.

摘要: Innovations in gene discovery and the analysis of expression are facilitating identification a growing number antigens that could potentially be targeted for immunotherapy tumors. Methods to reliably generate antigen-specific T-cell responses vitro would useful not only screen candidate immunogenicity prior embarking on vivo vaccination trials, but also lines or clones used directly adoptive approaches. Although many techniques have proven successful expanding ex effector cells from memory CD8(+) been primed vivo, methods high-avidity CTL naive repertoire well described. Various induction expansion healthy A2(+) donors were compared, using WT1 as model tumor-associated antigen which there is low frequency precursor T individuals. In contrast well-studied Melan-A/MART-1 (Melan-A) A2-restricted response, unusually high, WT1-specific appeared more dependent upon cell culture conditions. particular, primary stimulation with autologous peptide-loaded monocyte-derived DC generated 48 h (DC2d) was effective WT1-reactive populations than standard week-long protocol (DC7d). Adding supplemental IL-7 2 3 days after initiation cycle expanded within efficiently including cytokine beginning cycle. Following mature DC, subsequent restimulation PBMC stimulators at repeated DC. Using these techniques, specific an A()0201-restricted epitope nearly all normal tested. Such demonstrated capable recognizing lysing leukemic cells, will soon tested therapeutic activity clinical trials patients relapsed leukemia transplantation.