Isolation and Expansion of Tumor-Reactive Cytotoxic T-Cell Clones for Adoptive Immunotherapy

Attempts to treat patients with tumor-reactive cytotoxic T-lymphocytes (CTL) have been limited. This is due to the difficulty of isolating and expanding functionally active T-cells, which are present at extremely low frequencies in the peripheral blood. Recently developed multimers of the HLA-peptide complex mimic the natural ligand of the T-cell receptor and, therefore, fluorochrome-labeled multimers allow visualization and isolation of rare T-cells with defined specificity. Multimer-guided T-cell sorting permits the in vitro culture of antigen-specific T-cells as lines or clones. Cytolytic T-cells capable of recognizing HLA-peptide complexes endogenously processed by tumor cells are selected for further expansion, since lysis of tumor cells in vitro is a prerequisite for effective tumor elimination in vivo. The expansion of tumor-reactive CD8+ T-cells yields cell numbers sufficient for adoptive transfer. Tumor-reactive T-cells retain the functional activity in terms of cytolysis after expansion, encouraging their use in the immunotherapy of cancer patients.