Yu H W, Dong Y Y, Dang Y H
Department of Cardiology, the First Affiliated Hospital of Zhengzhou University, Zhengzhou 450000, China.
Zhonghua Yi Xue Za Zhi. 2020 May 5;100(17):1315-1319. doi: 10.3760/cma.j.cn112137-20190823-01876.
To investigate the expression of T cell immunoglobulin and mucin domain-containing protein 3 (TIM-3) in patients with chronic heart failure and its modulatory activity on T lymphocytes. Eighty-six patients with chronic heart failure (CHF group) who were hospitalized in Department of Cardiology, the First Affiliated Hospital of Zhengzhou University between January and October 2018 were enrolled in the study. Meanwhile, thirty-two healthy controls (HC group) who received healthy examination were also selected. Peripheral blood mononuclear cells were isolated, and TIM-3 expression of CD4(+)and CD8(+)T cells was investigated by flow cytometry. CD4(+)and CD8(+)T cells were purified, and were stimulated by anti-TIM-3 antibody. Interferon-γ (IFN-γ), interleukin (IL)-4, IL-10, IL-35, IL-17, and IL-22 expressions in the supernatants of cultured CD4(+)T cells and tumor necrosis factor-α (TNF-α) and IFN-γ expressions in the supernatants of cultured CD8(+) T cells were measured by enzyme-linked immunosorbent assay. mRNA expressions of T-bet, GATA-3, FoxP3, and RORγt in CD4(+)T cells and perforin and granzyme B in CD8(+)T cells were semi-quantified by real-time PCR. Student test or paired test was used for comparisons between the two groups. TIM-3(+) CD4(+) T cell percentage significantly increased in CHF group than that of HC group (3.47%±1.06% vs 0.92%±0.27%, 0.001). TIM-3(+)CD8(+)T cell percentage also notably elevated in CHF group compared to HC group (6.12%±1.91% vs 1.77%±0.63%, 0.001). CD4(+)T and CD8(+)T cells were dysfunctional in chronic heart failure. The levels of IFN-γ, IL-17, and IL-22 secreted by purified CD4(+) T cells significantly reduced in CHF group, while IL-10 and IL-35 expressions elevated in CHF group (all 0.05). The relative mRNA expression levels of T-bet and RORγt in CD4(+) T cells remarkably decreased in CHF group than those of HC group (all 0.01), while relative expression level of FoxP3 mRNA increased in CHF group (1.93±0.88 vs 0.97±0.28, 0.031). The levels of TNF-α and IFN-γ produced by purified CD8(+)T cells notably reduced in CHF group than those of HC group (all 0.05), and relative mRNA expression levels of perforin and granzyme B also decreased in CHF group (all 0.05). The levels of anti-TIM-3 antibody stimulation-produced IFN-γ, IL-17, and IL-22 increased (all 0.05) but IL-35 secretion reduced [(61±13) ng/ml vs (72±17) ng/ml, 0.029] by CD4(+)T cells in CHF group. The relative mRNA expression levels of T-bet and RORγt also elevated in response to anti-TIM-3 antibody stimulation (all 0.05). Anti-TIM-3 antibody stimulation promoted TNF-α and IFN-γ production by CD8(+)T cells in CHF group (all 0.01). The relative mRNA expression levels of perforin and granzyme B also increased (all 0.05). TIM-3 was increasingly expressed in T cells from patients with chronic heart failure, and might take part in the regulation of T cell dysfunction in chronic heart failure.
探讨慢性心力衰竭患者T细胞免疫球蛋白黏蛋白结构域3（TIM-3）的表达及其对T淋巴细胞的调节活性。选取2018年1月至10月在郑州大学第一附属医院心内科住院的86例慢性心力衰竭患者（CHF组）纳入研究。同时，选取32例接受健康体检的健康对照者（HC组）。分离外周血单个核细胞，采用流式细胞术检测CD4(+)和CD8(+)T细胞的TIM-3表达。纯化CD4(+)和CD8(+)T细胞，并用抗TIM-3抗体刺激。采用酶联免疫吸附测定法检测培养的CD4(+)T细胞上清液中干扰素-γ（IFN-γ）、白细胞介素（IL）-4、IL-10、IL-35、IL-17和IL-22的表达，以及培养的CD8(+)T细胞上清液中肿瘤坏死因子-α（TNF-α）和IFN-γ的表达。采用实时聚合酶链反应半定量检测CD4(+)T细胞中T-bet、GATA-3、FoxP3和RORγt的mRNA表达，以及CD8(+)T细胞中穿孔素和颗粒酶B的mRNA表达。两组间比较采用Student检验或配对检验。CHF组TIM-3(+)CD4(+)T细胞百分比显著高于HC组（3.47%±1.06% 对0.92%±0.27%，P<0.001）。CHF组TIM-3(+)CD8(+)T细胞百分比也显著高于HC组（6.12%±1.91% 对1.77%±0.63%，P<0.001）。慢性心力衰竭患者的CD4(+)T和CD8(+)T细胞功能失调。CHF组纯化的CD4(+)T细胞分泌的IFN-γ、IL-17和IL-22水平显著降低，而CHF组IL-10和IL-35表达升高（均P<0.05）。CHF组CD4(+)T细胞中T-bet和RORγt的相对mRNA表达水平显著低于HC组（均P<0.01），而CHF组FoxP3 mRNA相对表达水平升高（1.93±0.88对0.97±0.28，P=0.031）。CHF组纯化的CD8(+)T细胞产生的TNF-α和IFN-γ水平显著低于HC组（均P<0.05），CHF组穿孔素和颗粒酶B的相对mRNA表达水平也降低（均P<0.05）。CHF组CD4(+)T细胞经抗TIM-3抗体刺激后产生的IFN-γ、IL-17和IL-22水平升高（均P<0.05），但IL-35分泌减少[（61±13）ng/ml对（72±17）ng/ml，P=0.029]。抗TIM-3抗体刺激后，CHF组CD4(+)T细胞中T-bet和RORγt的相对mRNA表达水平也升高（均P<0.05）。抗TIM-3抗体刺激促进CHF组CD8(+)T细胞产生TNF-α和IFN-γ（均P<0.01）。穿孔素和颗粒酶B的相对mRNA表达水平也升高（均P<0.05）。TIM-3在慢性心力衰竭患者的T细胞中表达增加，可能参与慢性心力衰竭中T细胞功能失调的调节。
