Objective: To investigate the effect of Toll-like receptor 7 (TLR7) in CD8(+) T cells activity from patients with breast cancer. Methods: Thirty-three patients with breast cancer, twenty-three patients with benign breast tumor, and twenty healthy individuals were collected from The First Affiliated Hospital of Xinxiang Medical University between December 2017 and March 2018. Peripheral blood mononuclear cells (PBMCs) were isolated, and CD8(+) T cells were purified. TLR7 protein and mRNA relative expression in CD8(+) T cells was measured using flow cytometry and real-time PCR, respectively. mRNA relative expressions corresponding to perforin, granzyme B, and FasL in CD8(+) T cells were measured in response to TLR7 agonist stimulation. Direct/indirect contact co-culture system of CD8(+) T cells and breast cancer cell line MCF-7 was also used to assess cytolytic and noncytolytic function in response to TLR7 agonist CL097 stimulation. Results: The mean fluorescence intensity corresponding to TLR7 protein in CD8(+) T cells from breast cancer patients was 124.0±15.32, which was significantly down-regulated in comparison with benign breast tumor patients (255.5±54.91) and healthy individuals (261.9±68.65) (P<0.000 1). TLR7 mRNA relative level was also remarkably reduced in CD8(+) T cells from breast cancer patients (1.97±1.18) in comparison with benign breast tumor patients (4.84±1.01) and healthy individuals (4.75±1.40) (P<0.000 1). TLR7 agonist CL097 stimulation notably increased mRNA relative levels of perforin and granzyme B mRNA in CD8(+) T cells (P<0.01), but not elevated FasL mRNA (P>0.05).Furthermore, TLR7 agonist CL097 stimulation enhanced the cytolytic and noncytolytic function of CD8(+) T cells to MCF-7 cells, which presented as the elevation of target cell death and increase of interferon-γ production in direct and indirect contact co-culture system. Conclusion: TLR7 agonist promoted CD8(+) T cells function from breast cancer patients.
目的： 探讨Toll样受体7（Toll-like receptor 7, TLR7）对乳腺癌患者CD8(+)T细胞功能的影响。 方法： 入组2017年12月—2018年3月在新乡医学院第一附属医院就诊的33例乳腺癌患者、23例乳腺良性肿块患者和20例健康对照。分离外周血单个核细胞（PBMC），分选CD8(+)T细胞，应用流式细胞术和实时定量PCR法检测CD8(+)T细胞中TLR7蛋白表达和mRNA相对表达量。使用TLR7激动剂CL097刺激CD8(+)T细胞，检测穿孔素、粒酶B和FasL mRNA相对表达量的变化。建立CD8(+)T细胞和乳腺癌细胞系MCF-7的直接/间接接触共培养细胞，观察TLR7激动剂对CD8(+)T细胞杀伤和非杀伤功能的影响。 结果： TLR7蛋白在乳腺癌患者CD8(+)T细胞的平均荧光强度为124.0±15.3，显著低于乳腺良性肿块患者（255.5±54.9）和健康对照（261.9±68.7）（P<0.000 1）。TLR7 mRNA在乳腺癌患者CD8(+)T细胞中的相对表达量（1.97±1.18）亦显著低于乳腺良性肿块患者（4.84±1.01）和健康对照（4.75±1.40）（P<0.000 1）。TLR7激动剂CL097刺激可显著增加CD8(+)T细胞中穿孔素和粒酶B mRNA的相对表达量（P<0.01），但FasL mRNA的相对表达量在经CL097刺激和无CL097刺激的CD8(+)T细胞中的差异无统计学意义（P>0.05）。TLR7激动剂CL097刺激还可显著提升乳腺癌患者CD8(+)T细胞对MCF-7细胞的杀伤和非杀伤功能，表现为直接和间接接触共培养系统中靶细胞死亡比例的升高和干扰素-γ分泌的增加。 结论： TLR7激动剂可增强乳腺癌患者CD8(+)T细胞的功能。.
Keywords: Antineoplastic; Breast neoplasms; CD8-positive T-lymphocytes; Toll-like receptor 7.