Supplementary Materials and Methods, Figures 1 - 13, Table 1 from Activating and Propagating Polyclonal Gamma Delta T Cells with Broad Specificity for Malignancies

PDF file - 1405KB, Supplementary Materials and Methods. Supplementary Figures - (1) Tumor cell line culture conditions and media formulations (2) Co-culture conditions for gamma/delta T cells and designer aAPC (3) Intracellular cytokine production, Luminex, and neutralizing antibody cytolysis assays (4) Lentivirus packaging and transduction of CAOV3 cells Contents of the Supplemental Data include figures: (1) An example of the gating strategy for gamma/delta T-cell analyses (2) Schematic of DNA plasmid pLVU3G-effLuc-T2A-mKateS158A used to co-express enhanced firefly luciferase (effLuc) and mKate (3) Expression of activation markers CD38 and CD95 on propagated gamma/delta T cells (4) aAPC developed for co-culture with gamma/delta T cells to determine the impact of introduced co-stimulatory molecules (5) Expansion of UCB-derived gamma/delta T cells on aAPC with IL-2 and IL-21 (6) Surface expression of TCRdelta1 and TCRdelta2 chains on gamma/delta T cells derived from PBMC prior to propagation (7) Abundance of Vdelta and Vgamma mRNA species in gamma/delta T cells prior to ex vivo numeric expansion (8) Surface expression of TCRdelta1 and TCRdelta2 chains on gamma/delta T cells derived from UCB and propagated on aAPC with IL-2 and IL-21 (9) mRNA expression of shared Valpha/Vdelta alleles in gamma/delta T cells separated and then propagated on aAPC, IL-2, and IL-21 (10) Abundance of mRNA species coding for Vgamma chains in gamma/delta T-cell subsets (11) In vitro lysis of tumor cell line panel by polyclonal gamma/delta T cells (12) Specific lysis of hematological and solid tumor cells by Vdelta T-cell subsets (13) Immunophenotypes associated with PBMC-derived Vdelta T-cell subsets expanded on aAPC/IL-2/IL-21. Supplemental Table 1 - details the antibodies used in this study, where they were purchased, and their hybridoma clone.