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. 2021 Mar 22;16(3):e0248973. doi: 10.1371/journal.pone.0248973

Fig 4. Viral suppression activity by dual and triple anti-SIV CAR T cells.

Fig 4

(A) Dual anti-SIV-CAR transduction and viral suppression by dual anti-SIV-CAR T cells. Two different CAR vectors labeled with different reporter genes were mixed, coated on the same plates and transduced simultaneously. NGFR was stained with anti-NGFR-specific antibody and ZsGreen+NGFR+ cells were sorted for effector cells. Blue lines show suppression by ITS01 (CD4bs)+ITS06 (V1), grey lines show suppression by ITS01 (CD4bs)+ITS52 (V3) and black lines show suppression by ITS06.02 (V1)+ITS52 (V3) dual SIV-CAR T cells. Viral suppression on all tiers of SIV was assessed in duplicate. (B) Comparison of viral suppression efficacy based on valency. ITS01, ITS06.01 and/or CD4-MBL CAR were transduced to T cells separately or simultaneously. Single, dual or triple CAR T cells were sorted based on reporter gene or CAR expression. Sorted CAR T cells were co-cultured with SIVsmE660-FL14-infected cells in four replicates at E:T ratios of 1:1, 1:5 and 1:25. Percent reduction of p27-positive cells from CAR-negative samples were shown in the graph. E:T ratio at 0.04 was not performed for SIVmac239 due to shortage of CAR T cells for this assay.