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. 2020 Mar 15;10(10):4481–4489. doi: 10.7150/thno.41646

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TNF blockade attenuates AICD in human CD8 tumor-infiltrating lymphocytes. Fresh cell suspensions from surgical tumor specimens were activated with plate-bound anti-CD3 and soluble anti-CD28 mAbs for 4 days in the presence or absence of anti-TNF blocking agents. The type of tumor is color-coded and indicated in the figure legend (CRC: colorectal carcinoma, LmCRC: colorectal carcinoma liver metastasis, RCC: renal cell carcinoma, Endo: endometrial adenocarcinoma, Prost: prostate carcinoma). Apoptotic cell death was measured by surface Annexin V binding (A) and active caspase 3 intracellular staining (B) on tumor-infiltrating CD8 T cells. Data for A; anti-CD3/CD28 + etanercept (n = 9); anti-CD3/CD28 + infliximab (n = 7). Data for B; (n = 5). C, representative dot plots of phosphorylated gH2AX on FACs-gated CD8 T cells from an endometrial cancer in the cultures set up. CD8 T cells were pre-gated on Live+/CD45-/CD3+/CD4- . P values were calculated using a paired two-tailed t-test.