Chinese Journal of Schistosomiasis Control ›› 2021, Vol. 33 ›› Issue (6): 575-.

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Role of inducible costimulatory molecules (ICOS) and related cytokines in immune regulation of Echinococcus granulosus infections in mice

YANG Chen⁃chen1, ZHANG Ji⁃xiu2, WEI Qin3, JIANG Tao3*   

  1. 1 Department of Basic Medicine, School of Healthy and Nursing, Wuxi Taihu University, Wuxi, Jiangsu 214064, China; 2 Changji Vocational and Technical College, China; 3 Center for Laboratory Animals, Xinjiang Medical University, Urumqi, Xinjiang 830011, China
  • Online:2022-01-12 Published:2022-01-27

可诱导共刺激分子及相关细胞因子在细粒棘球蚴感染小鼠免疫调控中的作用

杨晨晨1,张继秀2,魏琴3,姜涛3*   

  1. 1无锡太湖学院健康与护理学院基础医学教研室(江苏 无锡 214064);2 昌吉职业技术学院;3新疆医科大学动物实验中心(新疆 乌鲁木齐 830011)
  • 作者简介:杨晨晨,女,博士,副教授。研究方向:棘球蚴病发病机制
  • 基金资助:
    新疆维吾尔自治区自然科学基金(2018D01C190)

Abstract: Objective To investigate the roles of inducible costimulatory molecules (ICOS) and related cytokines in the immune regulation of Echinococcus granulosus infections in mice. Methods Eighty BALB/c mice (weight 18-22 g) were divided into the control and infection groups, of 40 animals in each group. E. granulosus infection was modeled in mice by intraperitoneal injection of 10 000 protoscoleces per mouse. Serum levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT) and alkaline phosphatase (ALP) and peripheral interleukin⁃4 (IL⁃4) and IL⁃10 levels were measured 2, 8, 30, 60, 180 days post⁃infection. Mouse liver specimens were excised for hematoxylin⁃eosin (HE) staining and immunostaining, and ICOS expression was quantified in mouse liver specimens using quantitative real⁃time PCR (qPCR) assay. Results There were no significant differences in serum ALT (F = 12.082, P < 0.05), AST (F = 6.347, P < 0.05) or ALP levels (F = 52.186, P < 0.05) in mice 2, 8, 30, 60 and 180 days post⁃infection with E. granulosus. The serum ALT levels were significantly higher in the infection group than in the control group 2 [(61.72 ± 9.89) vs. (50.65 ± 4.67) U/L, P < 0.05] and 30 days post⁃infection [(80.61 ± 23.71) vs. (67.75 ± 9.79) U/L, P < 0.05], and the serum ALT levels were significantly higher in the infection group than in the control group 2 [(181.06 ± 60.61) vs. (115.58 ± 17.66) U/L, P < 0.05] and 180 days post⁃infection [(137.84 ± 29.01) vs. (108.05 ± 10.33) U/L, P < 0.05], while greater serum ALP levels were measured in the infection group than in the control group 2 [(162.90 ± 21.04) vs. (64.54 ± 5.99) U/L, P < 0.05], 8 [(176.36 ± 24.56) vs. (62.70 ± 9.21) U/L, P < 0.05] and 30 days post⁃infection [(138.86 ± 13.59) vs. (58.60 ± 5.28) U/L, P < 0.05]. A few inflammatory cells were seen in mouse liver in the infection group 30 days post⁃infection, and no apparent changes were found in the mouse hepatic structure 60 days post⁃infection. On day 180 post⁃infection, a large number of epithelium⁃like cells presented fibrotic growth in mouse liver in the cyst⁃infiltrating regions, with cuticula formation seen, and plenty of red cells were present in lesions and hepatocyte space. Positive ICOS expression was detected in mouse liver in the infection group, with ICOS⁃positive cells predominantly seen in the cytoplasm of the hepatocyte, and the ICOS expression increased over time. The relative ICOS mRNA expression was 2.732 ± 0.094 on day 180 post⁃infection, which was significantly greater than that on day 2 post⁃infection (0.746 ± 0.049). There were no significant differences in serum IL⁃4 or IL⁃10 levels at different time points after E. granulosus infections, while the serum IL⁃4 and IL⁃10 levels peaked in the infection group180 days and 60 days post⁃infection, respectively. Higher serum IL⁃4 levels were measured in the infection group than in the control group 8 [(22.50 ± 3.24) vs. (5.82 ± 0.49) pg/mL, P < 0.05], 30 [(15.49 ± 4.73) vs. (5.10 ± 1.38) pg/mL, P < 0.05], 60 [(36.93 ± 6.14) vs. (4.13 ± 1.19) pg/mL, P < 0.05] and 180 days post⁃infection [(198.35 ± 0.70) vs. (4.19 ± 0.98) pg/mL, P < 0.05], and higher IL⁃10 levels were measured in the infection group than in the control group 2 [(4.84 ± 1.91) vs. (2.11 ± 1.03) pg/mL, P < 0.05], 8 [(44.72 ± 14.63) vs. (3.16 ± 0.60) pg/mL, P < 0.05], 30 [(25.47 ± 8.00) vs. (3.83 ± 1.87) pg/mL, P < 0.05], 60 [(187.16 ± 60.44) vs. (3.69 ± 1.05) pg/mL, P < 0.05] and 180 days post⁃infection [(85.40 ± 7.15) vs. (3.25 ± 0.93) pg/mL, P < 0.05]. Conclusions High ICOS expression is present in the liver of mice with E. granulosus infections. The positive ICOS expression and immune activation levels increase with the time of E. granulosus infections, leading to aggravation of hepatocyte injury caused by inflammation.

Key words: Echinococcus granulosus, Inducible costimulatory molecule, Inflammatory factor, Interleukin 10, Interleukin 4, Immune regulation

摘要: 目的 探讨可诱导共刺激分子(inducible costimulatory molecule, ICOS)及相关细胞因子在细粒棘球蚴感染小鼠免疫调控中的作用。方法 80只BALB/c小鼠(体质量18 ~ 22 g)随机分为对照组和感染组,每组40只。按10 000个原头节/只腹腔注射制备细粒棘球蚴感染小鼠模型,腹腔接种2、8、30、60、180 d后,测定小鼠血清天冬氨酸氨基转移酶(AST)、丙氨酸氨基转移酶(ALT)、碱性磷酸酶(ALP)含量,检测小鼠外周血细胞因子白细胞介素10(IL⁃10)和IL⁃4含量。取小鼠肝脏进行HE染色和免疫组织化学染色,应用实时定量PCR(qPCR)技术检测小鼠肝组织中ICOS表达水平。结果 细粒棘球蚴感染后2、8、30、60、180 d,小鼠血清ALT、AST和ALP水平差异均有统计学意义(F = 12.082、6.347、52.186,P均< 0.05)。感染后2 [(61.72 ± 9.89) vs. (50.65 ± 4.67) U/L]、30 d [(80.61 ± 23.71) vs. (67.75 ± 9.79) U/L],感染组小鼠血清ALT水平显著高于对照组(P均< 0.05);感染后2 [(181.06 ± 60.61) vs. (115.58 ± 17.66) U/L]、180 d [(137.84 ± 29.01) vs. (108.05 ± 10.33) U/L],感染组小鼠血清AST水平显著高于对照组(P均< 0.05);感染后2 [(162.90 ± 21.04) vs. (64.54 ± 5.99) U/L]、8 [(176.36 ± 24.56) vs. (62.70 ± 9.21) U/L]、30 d[(138.86 ± 13.59) vs. (58.60 ± 5.28) U/L],感染组小鼠血清ALP水平显著高于对照组(P均< 0.05)。感染后30 d,感染组小鼠肝脏可见少量炎性细胞;感染后60 d,小鼠肝脏结构未见明显改变;感染后180 d,小鼠包囊浸润区肝脏可见大量上皮样细胞呈纤维化生长,生成角质层,在病灶区及肝细胞间隙有大量红细胞存在。感染组小鼠肝脏中ICOS呈阳性表达,阳性细胞主要位于肝细胞胞质中,ICOS表达水平随着感染时间延长而逐渐增强;ICOS mRNA在感染180 d后相对表达量为2.732 ± 0.094,明显高于感染后2 d(0.746 ± 0.049)。对照组小鼠血清IL⁃4、IL⁃10水平在细粒棘球蚴感染后不同时间点无显著变化;感染组小鼠血清IL⁃4、IL⁃10水平分别于感染后180 d和60 d达高峰。感染后8 [(22.50 ± 3.24) vs. (5.82 ± 0.49) pg/mL]、30 [(15.49 ± 4.73) vs. (5.10 ± 1.38) pg/mL]、60 [(36.93 ± 6.14) vs. (4.13 ± 1.19) pg/mL]、180 d [(198.35 ± 0.70) vs. (4.19 ± 0.98) pg/mL],感染组小鼠血清IL⁃4水平均显著高于对照组(P均< 0.05);感染后2 [(4.84 ± 1.91)vs.(2.11 ± 1.03) pg/mL]、8 [(44.72 ± 14.63)vs.(3.16 ± 0.60) pg/mL]、30 [(25.47 ± 8.00) vs. (3.83 ± 1.87) pg/mL]、60 [(187.16 ± 60.44) vs. (3.69 ± 1.05) pg/mL]、180 d [(85.40 ± 7.15) vs. (3.25 ± 0.93) pg/mL],感染组小鼠血清IL⁃10水平均显著高于对照组(P均< 0.05)。结论 细粒棘球蚴感染小鼠肝脏出现ICOS高表达;随感染时间的延长,ICOS阳性表达逐渐增强,免疫活化水平逐渐升高,导致免疫炎症引起的肝细胞损伤逐渐加重。

关键词: 细粒棘球蚴, 可诱导共刺激分子, 炎症因子, 白细胞介素10, 白细胞介素4, 免疫调节

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