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. 2018 Jun 6;13(6):e0198347. doi: 10.1371/journal.pone.0198347

Fig 1. Schematic representation of the FOLR1-CAR construct and the specific activity against target cells expressing FOLR1.

Fig 1

(A) A FOLR1-CAR gene consisting of a FOLR1-specific scFv, CD28 and CD3ζ was constructed. (B) The FOLR1-CAR vector was transfected into Jurkat cells, and the Myc tagged FOLR1 gene was transfected into K562 cells. FOLR1-CAR and FOLR1-Myc tag vectors were measured by western blot using CD3ζ and myc antibody, respectively. (C) FOLR1-CAR Jurkat cells were co-cultured with transfected K562 cells at an effector-target (E/T) ratio of 10:1 for 24 h. The levels of IL-2 were measured by enzyme-linked immunosorbent assay (ELISA). (D−E) The specific activity of Jurkat cells transfected with the FOLR-CAR vector was analyzed by co-culture with FOLR1-positive KB cells. (D) Expression of FOLR1 in KB cells was measured using FACS. (E) The levels of IL-2 secreted by FOLR1-CAR Jurkat cells were measured by ELISA after co-cultured with KB cells at an E/T ratio of 10:1 for 24 h. Experiments were repeated at least three times with similar results. The data are represented as the mean cytokine concentrations ± SD (pg/ml) from triplicate cultures. Statistical analysis was performed using the two-sided unpaired Student’s t test (***P<0.001).