中国寄生虫学与寄生虫病杂志

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吡喹酮治疗后日本血吸虫感染小鼠滤泡辅助性T细胞及其表面分子的研究

张玉梅1,2,王燕娟1 *,刘华1,姜岩岩1,徐馀信1,郑力1,3,胡媛1,沈玉娟1,曹建平1   

  1. 1中国疾病预防控制中心寄生虫病预防控制所,卫生部寄生虫病原与媒介生物学重点实验室,科技部国家级热带病国际联合研究中心,世界卫生组织热带病合作中心,上海 200025;2 滨州医学院病原生物学教研室,烟台 264003;3湖北省疾病预防控制中心,武汉 430000
  • 出版日期:2016-04-30 发布日期:2016-06-30

T Follicular Helper Cells and Related Molecules in Schistosoma japonicum Infected Mice after Praziquantel Treatment

ZHANG Yu-mei1,2, WANG Yan-juan1 *, LIU Hua1, JIANG Yan-yan1, XU Yu-xin1,ZHENG Li1,3, HU Yuan1, SHEN Yu-juan1, CAO Jian-ping1   

  1. 1 National Institute of Parasitic Diseases, Chinese Center for Disease Control and Prevention;Key Laboratory of Parasite and Vector Biology, Ministry of Health;National Center for International Research on Tropical Diseases, Ministry of Science and Technology;WHO Collaborating Centre for Tropical Diseases, Shanghai 200025, China;2 Department of Pathogen Biology, Binzhou Medical College, Yantai 264003, China;3 Center for Disease Control and Prevention of Hubei Province, Wuhan 430000, China)
  • Online:2016-04-30 Published:2016-06-30

摘要: 目的 观察日本血吸虫(Schistosoma japonicum)感染小鼠肝脏、脾脏的病变情况及经吡喹酮治疗后小鼠外周血及脾脏中滤泡辅助性T细胞(Tfh)及其表面分子的变化。 方法 将15只6~8周龄C57BL/6雌性小鼠随机分为吡喹酮治疗感染组(治疗组)、未治疗感染组(未治疗组)和未感染组,每组5只。治疗组和未治疗组每只小鼠经腹部皮肤感染日本血吸虫尾蚴20条;治疗组小鼠于感染后6周给予200 mg/(kg·d)吡喹酮灌胃治疗,连续3 d;未感染组和未治疗组不治疗。于治疗后4周解剖各组小鼠,观察小鼠肝脏和脾脏病变情况,计算减虫率和肝脏减卵率。采用流式细胞术检测各组小鼠外周血和脾脏Tfh占CD4+ T细胞的比例,检测其表面分子可诱导T细胞共刺激分子(ICOS)和程序性死亡受体1(PD-1)的表达情况。采用ELISA检测小鼠血清中可溶性虫卵抗原(SEA)特异性IgG抗体水平。 结果 与未治疗组比较,治疗组小鼠的肝脏和脾脏病变明显较轻,减虫率和肝脏减卵率分别为84.1%和69.1%(P<0.01)。治疗组、未治疗组和未感染组外周血和脾脏Tfh细胞的比例分别是14.7%~18.0%和15.6%~25.0%、13.7%~16.7%和12.4%~18.2%、2.5%~6.8%和4.9%~8.0%,治疗组和未治疗组明显高于未感染组(P<0.01),治疗组和未治疗组差异无统计学意义(P>0.05)。治疗组、未治疗组和未感染组外周血和脾脏中ICOS的表达水平分别为0.7%~1.1%和1.8%~6.8%、1.3%~3.2%和4.1%~7.0%、0.2%~0.3%和0.5%~0.8%,未治疗组高于治疗组和未感染组(P<0.01);治疗组脾脏ICOS的表达水平明显高于未感染组(P<0.01),而外周血的表达水平与未感染组差异无统计学意义(P>0.05)。治疗组、未治疗组和未感染组外周血和脾脏中Tfh细胞PD-1水平分别为0.5%~1.5%和4.5%~8.9%、0.8%~1.9%和4.1%~10.7%、0.4%~0.8%和1.2%~1.8%,未治疗组明显高于未感染组(P<0.01),治疗组与未治疗组差异无统计学意义(P>0.05)。吡喹酮治疗后4周,治疗组、未治疗组和未感染组的SEA特异性IgG水平(A450值)分别为2.015±0.061、1.969±0.038和0.139±0.128,治疗组与未治疗组差异无统计学意义(P>0.05)。 结论 吡喹酮治疗后,日本血吸虫感染小鼠肝脏、脾脏组织病变明显减轻,外周血和脾脏Tfh细胞ICOS的表达比未治疗组显著降低。

关键词: 日本血吸虫, 吡喹酮, 滤泡辅助性T细胞, 小鼠

Abstract: Objective To investigate the pathological changes of liver and spleen of mice infected with Schistosoma japonicum and the changes of T follicular helper(Tfh) cells and surface molecules after praziquantel treatment. Methods Fifteen female C57BL/6 mice (6-8 weeks) were randomly assigned into the praziquantel treated infection group (treated group), infection control group(untreated group) and uninfected group (n=5 in each group). The mice in the treated group and untreated group were each infected with 20 S. japonicum cercariae through the abdominal skin, and mice in the treated group were further administered with intragastric praziquantel [200 mg/(kg·d)] at week 6 post-infection for 3 consecutive days. Mice were sacrificed at week 4 after treatment to observe the morphological changes of liver and spleen and calculate the worm reduction rate and the liver egg reduction rate. The Tfh cell to CD4+ T cell ratio, as well as the expression of inducible T-cell costimulator (ICOS) and programmed cell death protein 1(PD-1) on peripheral blood and spleen, were determined by flow cytometry. Schistosome soluble egg antigen(SEA) specific IgG antibodies in serum were detected by ELISA. Results The pathological changes of liver and spleen in the treated group were less severe compared with those of the untreated group, with a worm reduction rate of 84.1% and liver egg reduction rate of 69.1%. Flow cytometry showed that the percent of Tfh cells in peripheral blood and spleen was significantly higher in the treated group(14.7%-18.0%, 15.6%-25.0%) and the untreated group(13.7%-16.7%, 12.4%-18.2%) than that in the uninfected group(2.5%-6.8%, 4.9%-8.0%), but there was no significant difference between the treated and untreated groups. The expression of ICOS in the peripheral blood and the spleen was significantly higher in untreated group(1.3%-3.2%, 4.1%-7.0%) than in the treated group(0.7%-1.1%, 1.8%-6.8%) and the uninfected group(0.2%-0.3%, 0.5%-0.8%)(P<0.01), The expression of ICOS in the spleen was significantly higher in the treated group than in the uninfected group(P<0.01), while this difference was not found for ICOS expression in the peripheral blood. The PD-1 expression in the peripheral blood and spleen was significantly higher in the untreated group(0.8%-1.9%, 4.1%-10.7%) than in the uninfected group(0.4%-0.8%, 1.2%-1.8%)(P<0.01), while there was no significant difference between the treated group(0.5%-1.5%, 4.5%-8.9%) and the untreated group(P>0.05). In addition, there was no significant difference in the level of SEA specific IgG between the treated group(2.015±0.061) and the untreated group(1.969±0.038) at 4 weeks after praziquantel treatment. Conclusion Praziquantel treatment can significantly alleviate the lesions of the liver and the spleen and decrease the ICOS expression by Tfh cells in the peripheral blood and spleen.

Key words: Schistosoma japonicum, Praziquantel, T follicular helper cells, Mouse