Fig. 1

Lack of specificity of the ERV-K-Env-specific TCR against OC cell lines. a. Diagram of the ELISpot assay design used in b-c. b. Representative images of developed wells for the level of background IFN-γ secretion (media), IFN-γ secretion in the presence of ES-2 cells (1:1 ES-2), and positive control of IFN-γ (PHA) for the ELISpot assay quantified in c. Number of spots in each well are indicated to the bottom right of each image. Images shown are for OM9.2 T cells. c. Number of IFN-γ spots per well normalized to the positive control (expressed as % of PHA) of untransduced T cells (black) and OM9.2 T cells (red) for the indicated conditions: media = background level of IFN-γ secretion from T cells, actin = negative control, various effector:target ratios of T cells (effectors) with HLA-A*03:01 OC cell lines (targets), PHA = positive control. Each donor is indicated by a different symbol. ELISpot conditions were plated in duplicate or triplicate as cell numbers allowed for n = 3 donors. Error bars represent SEM of all data points for all donors. The Wilcoxon rank sum test was used to compare the number of IFN-γ spots between untransduced and transduced T cells (indicated by *). The Kruskal Wallis test was used to compare the number of IFN-γ spots between each antigen condition for a given T cell type. Any Kruskal Wallis test that had p < 0.05 was followed by post-hoc pairwise Wilcoxon rank sum tests and adjusted for multiple comparisons (indicated by #). All statistical comparisons for TCR experiments are in Tables S1 and S2. *p < 0.05, **p < 0.005, ***p < 0.0005. ELISpot = Enzyme-linked immunosorbent spot; PHA = phytohemagglutinin; OC = ovarian cancer; SEM = standard error of the mean