中国寄生虫学与寄生虫病杂志

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晚期血吸虫病患者外周血NK细胞Tim-3分子的表达及其与肝纤维化指标的关系

武其文1,朱翔1,付夏1,杨进孙2,操治国3,浦春1 *   

  1. 1 皖南医学院第一附属医院检验科,芜湖 241001;2皖南医学院第一附属医院感染科,芜湖 241001;3 安徽省血吸虫病防治研究所,合肥 230061
  • 出版日期:2015-10-30 发布日期:2016-01-06

Expression of Tim-3 on Peripheral CD56+ NK Cells and Its Correlation with Liver Fibrosis in Patients with Advanced Schistosomiasis

WU Qi-wen1, ZHU Xiang1, FU Xia1, YANG Jin-sun2, CAO Zhi-guo3, PU Chun1 *   

  1. 1 Department of Laboratory; 2 Department of Infectious Diseases, the First Affiliated Hospital of Wannan Medical College, Wuhu 241001, China; 3 Anhui Provincial Institute of Schistosomiasis Control, Hefei 230061, China
  • Online:2015-10-30 Published:2016-01-06

摘要:

目的  探讨T细胞免疫球蛋白和黏蛋白结构域分子3(T cell immunoglobulin and mucin domain protein 3,Tim-3)在晚期血吸虫病患者外周血CD56+自然杀伤细胞(NK细胞)表面的表达及其与肝纤维化指标的关系。  方法  采用流式细胞术检测28例临床晚期血吸虫病患者和30例健康者CD56+NK细胞表面Tim-3的表达,应用全自动化学发光免疫分析仪检测肝纤维化血清学三型前胶原N端肽(PIIIP N-P)、层黏连蛋白(LN)、IV型胶原(CIV)和透明质酸(HA)等4项指标,ELISA检测血清中γ-干扰素(IFN-γ)和白细胞介素-4(IL-4)水平,Fibroscan仪检测肝硬度值(LSM)反映肝纤维化程度。统计分析Tim-3表达水平与IL-4和IFN-γ水平、肝脏纤维化程度以及肝纤维化血清指标的相关性。  结果  流式细胞分析结果显示,晚期血吸虫病患者外周血CD56+NK细胞表面Tim-3表达水平为(62.3±11.4)%,高于健康对照组的(52.1±6.5)%(P<0.01)。晚期血吸虫病患者和健康对照组的PIIIP N-P水平分别为(86.5±29.5) ng/ml和(22.0±7.8) ng/ml,LN分别为(49.3±21.5) ng/ml和(20.4±6.3) ng/ml,CIV分别为(67.5±22.3) ng/ml和(22.0±3.9) ng/ml,HA分别为(645.9±483.1) ng/ml和(54.7±27.7) ng/ml,两组各指标差异均具有统计学意义(P<0.05)。晚期血吸虫病患者和健康对照组IFN-γ水平分别为(93.9±20.1) ng/L和(107.7±24.6) ng/L,IL?鄄4分别为(46.6±11.8) ng/L和(28.9±8.9) ng/L,两组差异均具有统计学意义(P<0.05)。Spearman非参数相关分析结果显示患者CD56+NK细胞表达的Tim-3水平与患者肝纤维化指标LSM、PIIIP N-P、LN、CIV和IL-4水平呈正相关(r=0.528~0.834,P<0.01),与IFN-γ水平呈负相关(r=-0.501,P<0.01),而与HA水平无明显相关性(r=0.352,P>0.05)。  结论  晚期血吸虫病患者外周血CD56+NK细胞Tim-3表达呈上调,并与患者肝纤维化指标LSM、PIIIP N-P、LN、CIV和IL-4水平呈正相关。

关键词: Tim-3, 晚期血吸虫病, CD56+NK细胞, 肝纤维化

Abstract:

Objective To explore the expression of T cell immunoglobulin and mucin domain protein 3 (Tim-3) on CD56+ NK cells in peripheral blood and its correlation with liver fibrosis indicators in patients with advanced schistosomiasis. Methods Tim-3 expression on CD56+ NK cells from 28 patients with advanced schistosomiasis and 30 healthy controls was determined by flow cytometry. The serum levels of IFN-γ and IL-4 were detected by enzyme-linked immunosorbent assay (ELISA). Fibroscan analyzer was used for liver stiffness measurement (LSM)to determine the extent of liver fibrosis. Four serological indicators of liver fibrosis, collagen type III N-peptide (PIIIP N-P), Laminin (LN), collagen IV (CIV) and hyaluronic acid (HA) were measured by the Automated Chemiluminescence Immunoassay Analyzer. Results Flow cytometry analysis showed that Tim-3 expression on CD56+ NK cells in advanced schistosomiasis patients was (62.3±11.4)%, significantly higher than that (52.1±6.5)% (P<0.01). In patients with advanced schistosomiasis and the healthy controls, the levels of PIIIP N-P were (86.5±29.5) ng/mL and (22.0±7.8) ng/mL, LN (49.3±21.5) ng/mL and (20.4±6.3) ng/mL, CIV (67.5±22.3) ng/mL and (22.0±3.9) ng/mL, HA (645.9±483.1) ng/mL and (54.7± 27.7) ng/mL, respectively. There were significant differences in all these indicators between the two groups (P<0.05). The levels of IFN-γ were (93.9±20.1) ng/L and (107.7±24.6) ng/L, and those of IL-4 were (46.6±11.8) ng/L and (28.9±8.9) ng/L respectively in patients with advanced schisto-somiasis and the healthy controls, both with significant differences (P<0.05). Spearman nonparametric correlation analysis showed that Tim-3 expression on CD56+ NK cells in patients was positively correlated with the levels of LSM, PIIIP N-P, LN, CIV and IL-4 (r=0.528-0.834, P<0.01), but negatively with serum IFN-γ (r=-0.501, P<0.01). No correlation was found with the HA level (r=0.352, P>0.05). Conclusion The expression of Tim-3 on peripheral CD56+ NK cells increases in patients with advanced schistosomiasis compared with the healthy controls, and it positively correlates with the levels of LSM, PIIIP N-P, LN, CIV and IL-4, four indicators of liver fibrosis.

Key words: T cell immunoglobulin and mucin domain protein 3, Advanced schistosomiasis, CD56+NK cell, Liver fibrosis