Research Article
A Full GMP Process to Select and Amplify Epitope-Specific T Lymphocytes for Adoptive Immunotherapy of Metastatic Melanoma
Table 2
Validation runs for the selection and amplification of Melan-A- and MELOE-1- specific T lymphocytes from HLA-A2 melanoma patient-derived PBMC.
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The theoretical number of specific cells is estimated from the % of tetramer-positive cells in the sorted population. Sorting yields are calculated by dividing the number of rosetted T cells after sorting by the theoretical number of specific T cells in the sorted populations. Amplification factors are estimated from the final number of amplified T cells divided by the number of rosetted T cells seeded for amplification on feeder cells. Purity of sorted and amplified T cells is assessed by tetramer/CD8 double staining. % of TNF producing T cells among tetramer positive cells : reactivity of amplified T cells is assessed by tetramer/TNF double staining, after peptide activation. |