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. 2015 May 26;4(11):e1047582. doi: 10.1080/2162402X.2015.1047582

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Figure 2 . — Detection of antitumor effects in vitro. (A) Inhibition of BC cell growth was observed when these cells were treated with K-CARs from patients (157 and 108), using the Alamar Blue assay. (B) CTL assays were employed to determine the cytotoxicity of K-CAR toward BC cells at various ratios of effector to target. Significantly greater lysis was demonstrated in both cell lines using K-CAR from patient 108 (top panel) and 243 (bottom-left panel). Enhanced specific lysis was observed in MDA-MB-231 cells using K-CAR from patient 233 with CD4⁺ T depletion (CD4-K-CAR), compared to no depletion of CD4⁺ T cells (K-CAR; bottom-right panel). (C) The expression of HERV-K was determined in BC cell lines transduced with a shRNAenv or control shRNAc by immunoblot assays using 6H5 mAb. Expression of HERV-K protein was downregulated by approximately 70–80%, compared to the expression level of shRNAc transduced cells. (D) The specific lysis of K-CAR T cells obtained from patients 108 and 257 or normal female donors was reduced in BC target cells stably transduced with shRNAenv compared with shRNAc. This result indicates that specific lysis is antigen-dependent.

Figure 2. — Detection of antitumor effects in vitro. (A) Inhibition of BC cell growth was observed when these cells were treated with K-CARs from patients (157 and 108), using the Alamar Blue assay. (B) CTL assays were employed to determine the cytotoxicity of K-CAR toward BC cells at various ratios of effector to target. Significantly greater lysis was demonstrated in both cell lines using K-CAR from patient 108 (top panel) and 243 (bottom-left panel). Enhanced specific lysis was observed in MDA-MB-231 cells using K-CAR from patient 233 with CD4⁺ T depletion (CD4-K-CAR), compared to no depletion of CD4⁺ T cells (K-CAR; bottom-right panel). (C) The expression of HERV-K was determined in BC cell lines transduced with a shRNAenv or control shRNAc by immunoblot assays using 6H5 mAb. Expression of HERV-K protein was downregulated by approximately 70–80%, compared to the expression level of shRNAc transduced cells. (D) The specific lysis of K-CAR T cells obtained from patients 108 and 257 or normal female donors was reduced in BC target cells stably transduced with shRNAenv compared with shRNAc. This result indicates that specific lysis is antigen-dependent.