Department of Immunology, University of Tübingen, Tübingen, Germany.
Department of Urology, University Hospital Tübingen, Tübingen, Germany.
Cancer Immunol Res. 2019 Nov;7(11):1891-1899. doi: 10.1158/2326-6066.CIR-19-0146. Epub 2019 Sep 4.
Drugs targeting the programmed cell death protein 1 (PD-1) pathway are approved as therapies for an increasing number of cancer entities, including renal cell carcinoma. Despite a significant increase in overall survival, most treated patients do not show durable clinical responses. A combination of checkpoint inhibitors could provide a promising improvement. The aim of the study was to determine the most promising checkpoint blockade combination for renal cell carcinoma patients. Tumor-infiltrating lymphocytes (TIL) and autologous peripheral blood mononuclear cells (PBMC) were isolated from patients undergoing surgery for primary tumors. Cells were stained for multicolor flow cytometry to determine the (co)expression of five inhibitory receptors (iR), PD-1, LAG-3, Tim-3, BTLA, and CTLA-4, on T-cell populations. The function of these TILs was assessed by intracellular cytokine staining after stimulation in the presence or absence of PD-1 ± LAG-3 or Tim-3-specific antibodies. Although the percentage of iR T cells was low in PBMCs, both CD4 and CD8 T cells showed increased frequencies of PD-1, LAG-3, and Tim-3 cells on TILs. The most frequent iR combination was PD-1 and LAG-3 on both CD4 and CD8 TILs. Blockade of PD-1 resulted in significant LAG-3, but not Tim-3, upregulation. The dual blockade of PD-1 and LAG-3, but not PD-1 and Tim-3, led to increased IFNγ release upon stimulation. Together, these data suggest that dual blockade of PD-1 and LAG-3 is a promising checkpoint blockade combination for renal cell carcinoma.
针对程序性死亡蛋白 1（PD-1）通路的药物已被批准用于治疗越来越多的癌症实体，包括肾细胞癌。尽管总体生存率显著提高，但大多数接受治疗的患者并未表现出持久的临床反应。联合使用检查点抑制剂可能会提供有前途的改善。本研究旨在确定最有前途的肾细胞癌患者的检查点阻断联合治疗方案。从接受原发性肿瘤手术的患者中分离肿瘤浸润淋巴细胞（TIL）和自体外周血单核细胞（PBMC）。通过多色流式细胞术对细胞进行染色，以确定 T 细胞群中五种抑制性受体（iR）、PD-1、LAG-3、Tim-3、BTLA 和 CTLA-4 的（共）表达。在存在或不存在 PD-1±LAG-3 或 Tim-3 特异性抗体的情况下，通过刺激后细胞内细胞因子染色评估这些 TIL 的功能。尽管 PBMC 中 iR T 细胞的百分比较低，但 CD4 和 CD8 T 细胞在 TIL 上均显示出 PD-1、LAG-3 和 Tim-3 细胞的频率增加。最常见的 iR 组合是 CD4 和 CD8 TIL 上的 PD-1 和 LAG-3。PD-1 阻断导致 LAG-3 的显著上调，但 Tim-3 没有上调。PD-1 和 LAG-3 的双重阻断，而不是 PD-1 和 Tim-3 的双重阻断，导致在刺激后 IFNγ 的释放增加。这些数据表明，PD-1 和 LAG-3 的双重阻断是肾细胞癌有前途的检查点阻断联合治疗方案。
