丰宫并殖吸虫感染诱导大鼠Th1/Th2免疫应答的偏移模式研究  

Study on patterns of the Th1/Th2 immune response induced by Paragonimus proliferus infection in rats

作　　者：李生浩 杜映荣 武昆利 李俊义 杨永锐 常国楫 华丽娟 徐晶晶 马志强 张燕玲 李海雯 王文林[2] LI Sheng-hao;DU Ying-rong;WU Kun-li;LIJun-yi;YANG Yong-rui;CHANG Guo-ji;HUA Li-juan;XU Jing-jing;MA Zhi-qiang;ZHANG Yan-ling;LI Hai-wen;WANG Wen-lin(Department Of Hepatology,The Third People’s Hospital of Kunming,Kunming 650041,China;School of Basic Medicine,Kunming Medical University)

机构地区：[1]昆明市第三人民医院肝病科,云南昆明650041 [2]昆明医科大学

出　　处：《中国病原生物学杂志》2021年第2期171-176,共6页Journal of Pathogen Biology

基　　金：云南省地方本科高校基础研究联合专项基金项目(No.2018FH001-087);昆明市科技保障民生发展计划基金项目(昆科计字2019-1-S-25318000000808);昆明市卫健委卫生科研课题(No.2020-03-08-113)。

摘　　要：目的明确丰宫并殖吸虫感染后宿主(SD大鼠)Th1/Th2免疫偏移模式。方法自丰宫并殖吸虫流行区采集溪蟹,分离丰宫并殖吸虫后尾蚴,采用腹壁皮下注射的方法建立丰宫并殖吸虫感染大鼠动物模型,分别在0、3、7、14、28、70 d处死大鼠,取血,分离血清,采用双抗体夹心ELISA法检测I型免疫反应(IL-1、TNF-α、INF-γ)和II型免疫反应(IL-4、IL-5、IL13)相关细胞因子,结合肺组织病变情况,分析丰宫并殖吸虫大鼠肺吸虫病的Th1/Th2免疫偏移模式。结果大鼠感染丰宫并殖吸虫后,血清Th1型细胞因子(IL-1、TNF-α、INF-γ)水平迅速上升,感染7 d左右达到高峰,随后呈逐渐下降趋势,其中感染3 d、7 d与0 d、14 d、28 d、70 d时比较[IL-1:(3、7 d)vs.(0、14、28、70 d)=(25.25±2.26、23.00±8.38)pg/ml vs.(17.34±2.76、15.89±4.44、10.34±2.90、10.82±6.05)pg/ml;IFN-γ:(3、7 d)vs.(0、14、28、70 d)=(279.58±51.43、276.29±54.64)pg/ml vs.(173.64±43.36、190.74±34.10、142.57±35.10、170.56±26.53)pg/ml;TNF-α:(3、7 d)vs.(0、14、28、70 d)=(49.39±8.03、54.78±13.64)pg/ml vs.(33.10±4.75、37.64±19.53、25.01±11.91、23.12±9.05)pg/ml],差异均有统计学意义(均P<0.05)。Th2型细胞因子(IL-4、IL-5、IL-13)呈逐渐上升趋势,14 d或28 d时达到高峰,之后逐渐下降,70 d时下降至感染早期水平,其中14 d、28 d与0 d、3 d、7 d时比较[IL-4:(14、28 d)vs.(0、3、7、70 d)=(20.27±3.31、19.71±2.41)pg/ml vs.(10.77±2.58、9.90±3.62、12.92±3.65、11.72±2.42)pg/ml;IL-5:(14、28 d)vs.(0、3、7、70 d)=(18.68±2.46、19.53±2.65)pg/mL vs.(9.60±1.58、9.38±2.24、10.76±2.85、8.10±1.69)pg/ml;IL-13:(14、28 d)vs.(0、3、7、70 d)=(13.57±0.94、13.38±1.10)pg/ml vs.(6.00±1.33、7.09±1.18、8.68±2.03、7.12±0.90)pg/ml],差异均有统计学意义(均P<0.05)。肺部特征与细胞因子水平变化规律相一致。结论在丰宫并殖吸虫感染大鼠,Th1型及Th2型免疫反应共同存在,综合作用于宿主。早期(3 d和7 d)以I�Objective To identify patterns of the Th1/Th2 immune response in a host(SD rats)after infection with Paragonimus proliferus.Methods Freshwater crabs were collected from natural foci of P.proliferus to isolate metacercariae of this parasite species.Six-week-old male SD rats with a weight of 180-220 g were divided into 6 groups that were sacrificed on Day 0,3,7,14,28,and 70(8 rats per group).Rats sacrificed on Day 0 were not infected.The latter 5 groups(Day 3,7,14,28,and 70)were used to establish an animal model of P.proliferus infection via subcutaneous injection of metacercariae into the abdominal wall(8 metacercariae per rats).Serum was collected from rats on Day 3,7,14,28,and 70 post-infection(PI).Serum from the group sacrificed on Day 0 was collected at the beginning of the experiment.Levels of Th1 cytokines(IL-1,TNF-α,and IFN-γ)and Th2 cytokines(IL-4,IL-5,and IL-13)were determined using ELISA.In conjunction with lung lesions,Th1/Th2 immune response patterns were evaluated.Results The levels of Th1 cytokines(IL-1,TNF-α,and IFN-γ)increased rapidly in serum from rats infected with P.proliferus,the levels differed significantly on Day 3 and 7 compared to the levels on Day 0(P<0.05),and levels peaked on Day 7 and then gradually decreased[IL-1:Day 3 and 7 vs.Day 0,14,28,and 70=25.25±2.26 and 23.00±8.38 pg/ml vs.17.34±2.76,15.89±4.44,10.34±2.90,and 10.82±6.05 pg/ml;IFN-γ:Day 3 and 7 vs.Day 0,14,28,and 70=279.58±51.43 and 276.29±54.64 pg/ml vs.173.64±43.36,190.74±34.10,142.57±35.10,and 170.56±26.53 pg/ml;TNF-α:Day 3 and 7 vs.Day 0,14,28,and 70=49.39±8.03 and 54.78±13.64 pg/ml vs.33.10±4.75,37.64±19.53,25.01±11.91,and 23.12±9.05 pg/ml].Th2 cytokines(IL-4,IL-5,and IL-13)tended to increase gradually.Levels during the early stages of infection(Day 3 or 7)did not differ significantly compared to levels on Day 0,but they peaked on Day 14 or 28 and were significantly higher than those on Day 0,3,or 7(P<0.05).Levels on Day 70 decreased to levels during the early stages[IL-4:Day 14 and 28 vs.Day 0,3,7,

关 键 词：并殖吸虫病 丰宫并殖吸虫 Th1/Th2免疫反应 细胞因子 

分 类 号：R383.2[医药卫生—医学寄生虫学]