Additional file 1: Table S1. of RNA-transfection of γ/δ T cells with a chimeric antigen receptor or an α/β T-cell receptor: a safer alternative to genetically engineered α/β T cells for the immunotherapy of melanoma
Creators: name: Harrer, Dennis
name: Simon, Bianca
name: Shin-Ichiro Fujii
name: Shimizu, Kanako
name: Ugur Uslu
name: Schuler, Gerold
name: Gerer, Kerstin
name: Hoyer, Stefanie
name: Dörrie, Jan
name: Schaft, Niels
Publisher: name: Figshare
Year: 2017
Subjects: subject: Biochemistry
subject: Space Science
subject: Medicine
subject: Cell Biology
subject: Molecular Biology
subject: Immunology
subject: FOS: Clinical medicine
subject: FOS: Clinical medicine
subject: Developmental Biology
subject: Cancer
subject: Hematology
subject: 110309 Infectious Diseases
subject: FOS: Health sciences
subject: FOS: Health sciences
subject: 60506 Virology
subject: FOS: Biological sciences
subject: FOS: Biological sciences
subject: Computational Biology

description: Cellular composition of different cell populations pre and post expansion based on double stainings for γ/δ and CD3, and for γ/δ and CD8. Figure S1. Expression of a gp100/HLA-A2-specific TCR by different T-cell populations after electroporation. Zoledronic acid (ZA)-activated PBMC, OKT3-activated PBMC, OKT3-stimulated MACS-isolated CD8+ T cells, and OKT3-stimulated MACS-isolated γ/δ T cells were expanded as described in Fig. 1. After 10–11 days, these cells were electroporated with RNA coding for the gp100/HLA-A2-specific TCR or with RNA encoding the MCSP-specific CAR. After receptor transfer, T cells were rested for one day and subsequently cryopreserved. After thawing, TCR expression was detected using a PE-conjugated MHC-Dextramer HLA-A*0201/YLEPGPVTV (black lines). CAR-transfected T cells served as negative controls (neg.; filled grey histograms). Presented histograms are representatives out of three independent experiments. Table S2. Statistical analysis corresponding to Fig. 2g-j. Table S3. Statistical analysis corresponding to Fig. 3. Figure S2. Zoledronic acid-expanded γ/δ T cells retain their cytokine secretory capacity after depletion of γ/δ− cells. a Donor-derived PBMC were expanded with ZA (PBMC + ZA) as explained above (Fig. 1). Following 10 days of expansion, untouched γ/δ T cells were isolated from an aliquot of stimulated cells via negative selection using the TCR γ/δ T Cell Isolation Kit (after depletion). Subsequently, a γ/δ and CD3 double staining was employed to flow-cytometrically verify the successful depletion procedure. b + c On day 11, negatively isolated γ/δ T cells (after depletion, grey bars) and the remaining ZA-expanded T cells (black bars) were electroporated with RNA coding for the gp100/A2-specific TCR (b) or with RNA encoding the MCSP-specific CAR (c). T cells electroporated without RNA (mock) served as controls (b + c). Antigen-specific cytokine secretion was determined as described above (Fig. 3). Data represent means ± SEM from 4 independent experiments. P values calculated by unpaired Student’s t test are presented in Table S4. Table S4. Statistical analysis corresponding to Figure S2. b, c. Table S5. Statistical analysis corresponding to Fig. 5a, b. Table S6. Statistical analysis corresponding to Fig. 5c, d. Table S7. Statistical analysis corresponding to Fig. 6. (PDF 291 kb)