Fig. 2

Exposure to febrile temperature potentiates the production of IFN-γ. (A) Representative images illustrating a FACS workflow for the detection of IFN-γ positive T cells. Cells derived from donors’ buffy coats (BC) were activated at 37 ^oC or 39.5 ^oC and then co-cultured with STEAP1 antigen-expressing target cells. See C for the corresponding quantification. (B) Representative images illustrating a FACS workflow for the detection of IFN-γ positive CAR-T cells. Expression of the CAR was determined by staining for the marker protein RQR8, which comprises a CD34 epitope. Cells derived from donors’ buffy coats were activated at 37 ^oC or 39.5 ^oC and then co-cultured with STEAP1 antigen-expressing target cells. See D for the corresponding quantification. (C) The quantification of A, showing the percentage of IFN-γ positive non-transduced (NT-T) control T cells and CAR-T cells at the indicated temperatures. Colors indicate individual donors. Note that both NT-T cells and CAR-T cells exposed to 39.5 ºC had the tendency to overproduce IFN-γ. (D) The quantification of B, showing the percentage of IFN-γ positive CAR-T cells only. Colors indicate individual donors