Imai Yuichi, Hasegawa Kosei, Matsushita Hirokazu, Fujieda Nao, Sato Sho, Miyagi Etsuko, Kakimi Kazuhiro, Fujiwara Keiichi
Department of Gynecologic Oncology, Saitama Medical University International Medical Center, Hidaka-shi, Saitama 350-1298, Japan.
Gynecologic Oncology Translational Research Unit, Project Research Division, Research Center for Genomic Medicine, Saitama Medical University, Hidaka-shi, Saitama 350-1298, Japan.
Oncol Lett. 2018 May;15(5):6457-6468. doi: 10.3892/ol.2018.8101. Epub 2018 Feb 21.
Expression of immune checkpoint molecules, including programmed cell death protein-1 (PD-1), has been reported on T cells in various types of cancer. However, the expression status of these molecules in the tumor microenvironment of epithelial ovarian cancer (EOC) has not yet been studied. A total of 54 cases of malignant ascites from patients with EOC were analyzed in the present study. The expression of PD-1, lymphocyte-activation gene-3 (LAG-3), T-cell immunoglobulin and mucin-domain containing-3 (TIM-3) and B and T lymphocyte attenuator (BTLA) on cluster of differentiation (CD)4 and CD8 T cells in malignant EOC ascites were investigated using multicolor flow cytometric analysis. The expression of PD-L1 in tumor cells, PD-L2 in HLA-DR-positive cells and galectin-9 in ascitic fluid was also analyzed. In addition, cytokine profiling of ascitic fluid was performed to understand the immune microenvironment of EOC. PD-1, LAG-3 TIM-3, and BTLA were expressed on 65.8, 10.6, 4.3 and 37.6% of CD4 T cells, and on 57.7, 5.0, 4.9 and 15.7% of CD8 T cells, respectively. Programmed cell death protein-1 (PD-1), LAG-3 and BTLA were more frequently expressed on CD4 compared with CD8 T cells. The co-expression of immune checkpoints was further investigated and results indicated that 39 (72.2%) and 37 patients (68.5%) expressed multiple immune checkpoints on CD4 T cells and CD8 T cells, respectively. In addition, lower levels of TNF-α and interleukin-6 in ascitic fluid were significantly associated with multiple immune checkpoint expression on CD8 T cells. The present findings indicated that multiple immune checkpoint molecules were expressed on T cells in the EOC tumor microenvironment and the results may suggest the significance of simultaneous blockade of immune checkpoints to control EOC.
据报道，在各类癌症中，包括程序性细胞死亡蛋白1（PD-1）在内的免疫检查点分子在T细胞上均有表达。然而，这些分子在上皮性卵巢癌（EOC）肿瘤微环境中的表达情况尚未得到研究。本研究共分析了54例EOC患者的恶性腹水。采用多色流式细胞术分析了恶性EOC腹水中分化簇（CD）4和CD8 T细胞上PD-1、淋巴细胞激活基因3（LAG-3）、含T细胞免疫球蛋白和粘蛋白结构域3（TIM-3）以及B和T淋巴细胞衰减器（BTLA）的表达情况。还分析了肿瘤细胞中PD-L1、HLA-DR阳性细胞中PD-L2以及腹水中半乳凝素-9的表达情况。此外，对腹水进行细胞因子谱分析以了解EOC的免疫微环境。PD-1、LAG-3、TIM-3和BTLA分别在65.8%、10.6%、4.3%和37.6%的CD4 T细胞上表达，在57.7%、5.0%、4.9%和15.7%的CD8 T细胞上表达。与CD8 T细胞相比，程序性细胞死亡蛋白1（PD-1）、LAG-3和BTLA在CD4 T细胞上的表达更为频繁。进一步研究了免疫检查点的共表达情况，结果表明，分别有39例（72.2%）和37例（68.5%）患者的CD4 T细胞和CD8 T细胞表达多种免疫检查点。此外，腹水中较低水平的肿瘤坏死因子-α和白细胞介素-6与CD8 T细胞上多种免疫检查点的表达显著相关。本研究结果表明，EOC肿瘤微环境中的T细胞上表达多种免疫检查点分子，这些结果可能提示同时阻断免疫检查点对控制EOC的重要性。
