Figure 5.

The cytokines secreted by myeloid cells enhanced sCAR-T-induced endothelial activation. (A) Schematic presentation of CAR-T/Nalm6/PBMC or CAR-T/Nalm6/CD14+ cells co-culture in Transwell. Details refer to the information in the Materials and Methods. (B) The cytokine profiles of co-cultured supernatants were determined by multi-analyte flow assay. (C) The mRNA levels of endothelial activation-associated markers were determined by RT-PCR. GAPDH was taken as the housekeeping gene and data was expressed as fold changes relative to control. n = 3. (D) The protein expression of E-selectin, VCAM1, and ICAM1 was determined by flow cytometry. n = 3. (E) The protein expression of TF in HUVEC was determined by western blot and GAPDH was taken as the loading control. (F) The concentration of Ang2 secreted by HUVEC was assessed by ELISA. n = 3. (G) Confluent HUVEC cultured in Transwell were incubated with sCAR-T supplemented with or without adalimumab for 12h. The permeability of endothelial monolayer was determined by EBA. * represents p < 0.05, ** represents p < 0.01, *** represents p < 0.001, and **** represents p < 0.0001. ns represents not significant. All data were representative of at least three independent experiments.