New Approaches for Monitoring CTL Activity in Clinical Trials

摘要: We have developed a modification of the ELISPOT assay that measures Granzyme B (GrB) release from cytotoxic T lymphocytes (CTLs). The GrB is superior alternative to 51Cr-release since it significantly more sensitive and provides an estimation effector cell frequency. Additionally, unlike IFN-γ assay, directly cytolytic protein. report can be utilized measure ex vivo antigen-specific cytotoxicity peripheral blood mononuclear cells (PBMCs) cancer patients vaccinated with peptide-based vaccine. compare reactivity patients’ PBMCs in ELISPOT, tetramer, chromium (51Cr)-release assays. Differences immune response over all assays tested were found between patients, four patterns observed. Reactivity was closely associated than tetramer or gLISPOT also optimized responses against autologous primary tumor patients. A perforin adapted evaluate peptide-stimulated PMBCs melanoma Modifications described this chapter allow comprehensive evaluation low-frequency tumor-specific CTLs their specific functions provide valuable insight into vaccine trials.