细粒棘球蚴早期感染对小鼠T细胞亚群的影响
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摘要
目的观察细粒棘球蚴感染早期,小鼠体内脾细胞中T细胞亚群及相关细胞因子表达的改变。方法以细粒棘球蚴感染Balb/c小鼠后收集细胞用FCM检测Th1、Th2、Treg、Th9细胞;同时用qRT-PCR检测IL-9、IL-10、IFN-γ、IL-4、PU.1和TGF-β的mRNA水平表达量;用ELISA检测血清中IL-9、IL-10、IFN-γ、IL-4、TGF-β和IL-25的含量。结果 FCM检测原头蚴组不同时间点Th1、Th2、Treg、Th9细胞的比例,差异均有统计学意义(P<0.05);qRT-PCR检测IL-9、IL-10、IFN-γ、IL-4、PU.1和TGF-β的mRNA水平表达量在感染后升高,与对照组相比差异有统计学意义(P<0.05);ELISA检测血清中IL-9、IL-10、IFN-γ、IL-4、TGF-β和IL-25的含量在感染后升高,与对照组相比差异有统计学意义(P<0.05)。结论原头蚴能刺激小鼠体内脾细胞向Th1、Th2、Treg、Th9细胞分化且上调相应细胞因子的分泌,且Th9与Th2、Treg之间呈正相关,提示这些亚群细胞可能在包虫病感染的免疫逃逸中发挥一定作用。
This study was designed to observe the effect of Echinococcus granulosus on T subsets and relative cytokines in mouse spleen cells in vivo. Balb/c mice were infected with Echinococcus granulosus, and the percentage of Th subsets was detected by flow cytometry. The mRNA levels of IL-9, IL-10, FN-γ, IL-4, PU.1 and TGF-βexpression were detected using qRT-PCR. Then the levels of IL-9, IL-10, IFN-γ, IL-4, TGF-β and IL-25 in serum were detected by ELISA. Data showed in the early stages of Echinococcus granulosus infection in Balb/c mice, the levels of Th1, Th2, Treg, and Th9 cells were significant different at different time points(P<0.05);the mRNA levels of IL-9, IL-10, IFN-γ, IL-4, PU.1 and TGF-β were significantly diffeent increased after infection(P<0.05). And the levels of IL-9, IL-10, IFN-γ, IL-4,TGF-β and IL-25 in serum were significantly increased, as compared to the control group(P<0.05).Taken together, the protoscolex can stimulate spleen cells to differentiate to Th1, Th2, Treg, and Th9 cells,and up-regulate relative cytokines, among which, Th9 is positively related with Th2 and Treg, suggesting that these subsets may have some impaction on the immune invasion of hydatid.
This study was designed to observe the effect of Echinococcus granulosus on T subsets and relative cytokines in mouse spleen cells in vivo. Balb/c mice were infected with Echinococcus granulosus, and the percentage of Th subsets was detected by flow cytometry. The mRNA levels of IL-9, IL-10, FN-γ, IL-4, PU.1 and TGF-βexpression were detected using qRT-PCR. Then the levels of IL-9, IL-10, IFN-γ, IL-4, TGF-β and IL-25 in serum were detected by ELISA. Data showed in the early stages of Echinococcus granulosus infection in Balb/c mice, the levels of Th1, Th2, Treg, and Th9 cells were significant different at different time points(P<0.05);the mRNA levels of IL-9, IL-10, IFN-γ, IL-4, PU.1 and TGF-β were significantly diffeent increased after infection(P<0.05). And the levels of IL-9, IL-10, IFN-γ, IL-4,TGF-β and IL-25 in serum were significantly increased, as compared to the control group(P<0.05).Taken together, the protoscolex can stimulate spleen cells to differentiate to Th1, Th2, Treg, and Th9 cells,and up-regulate relative cytokines, among which, Th9 is positively related with Th2 and Treg, suggesting that these subsets may have some impaction on the immune invasion of hydatid.
引文
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[24]陈杰,管俊昌.Th17/Treg细胞的免疫失衡与病原体的感染[J].蚌埠医学院学报,2018,43(10):1305-1308.
[2]Siracusano A,Delunardo F,Teggi A,et al.Host-parasite relationship in cystic echinococcosis:an evolving story[J].Clin Dev Immunol,2012,2012:639362.
[3]Carmena D,Sanchez-Serrano LP,Barbero-Martinez I.Echinococcus granulosus infection in Spain[J].Zoonoses Public Health,2008,55(3):156-165.
[4]Craig PS,Hegglin D,Lightowlers MW,et al.Echinococcosis:Control and Prevention[J].Adv Parasitol,2017,96:55-158.
[5]Agudelo Higuita NI,Brunetti E,Mccloskey C.Cystic Echinococcosis[J].J Clin Microbiol,2016,54(3):518-523.
[6]Bhutani N,Kajal P.Hepatic echinococcosis:A review[J].Ann Med Surgery,2018,36:99-105.
[7]Jankovic D,Liu Z,Gause WC.Th1-and Th2-cell commitment during infectious disease:asymmetry in divergent pathways[J].Trends Immunol,2001,22(8):450-457.
[8]Licona Limon P,Arias Rojas A,Olguin Martinez E.IL-9and Th9 in parasite immunity[J].Seminars Immunopathol,2017,39(1):29-38.
[9]Tang J,Li Y,Sang Y,et al.LncRNA PVT1 regulates triplenegative breast cancer through KLF5/beta-catenin signaling[J].Oncogene,2018,37(34):4723-4734.
[10]Diaz A.Immunology of cystic echinococcosis(hydatid disease)[J].Br Med Bull,2017,124(1):121-133.
[11]Chen X,Zhang J,Feng X,et al.Humoural immune response and pathological analysis in patients with false immune diagnosis of cystic echinococcosis[J].Parasite Immunol,2014,36(4):170-176.
[12]潘伟,李向阳,孙芬芬,等.细粒棘球蚴慢性感染阶段小鼠血清中炎症因子的变化意义[J].中华血吸虫病防治杂志,2016,28(5):527-529.
[13]Mourglia Ettlin G,Cucher M,Arbildi P,et al.Natural and induced antibodies contribute to differential susceptibility to secondary cystic echinococcosis of Balb/c and C57Bl/6mice[J].Immunobiology,2016,221(1):103-115.
[14]Mourglia Ettlin G,Marques JM,Chabalgoity JA,et al.Early peritoneal immune response during Echinococcus granulosus establishment displays a biphasic behavior[J].PLoS Negl Trop Dis,2011,5(8):e1293.
[15]Dempster RP,Berridge MV,Harrison GB,et al.Echinococcus granulosus:development of an intermediate host mouse model for use in vaccination studies[J].Int JParasitol,1991,21(5):549-554.
[16]Zhang Y,Zhang Y,Gu W,et al.Th1/Th2 cell′s function in immune system[J].Adv Exp Med Biol,2014,841:45-65.
[17]Zhang Y,Zhang Y,Gu W,et al.TH1/TH2 cell differentiation and molecular signals[J].Adv Exp Med Biol,2014,841:15-44.
[18]杨志刚,文瑞婷,郑桂仙,等.调节性T细胞及Th1/Th2细胞因子在急性白血病中的表达及意义[J].现代肿瘤医学,2018,26(22):3641-3644.
[19]Sheng J,Chen W,Zhu HJ.The immune suppressive function of transforming growth factor-beta(TGF-beta)in human diseases[J].Growth factor,2015,33(2):92-101.
[20]Barbi J,Pardoll D,Pan F.Treg functional stability and its responsiveness to the microenvironment[J].Immunol Rev,2014,259(1):115-139.
[21]Yoshimura A,Muto G.TGF-beta function in immune suppression[J].Curr Top Microbiol Immunol,2011,350:127-147.
[22]刘乃国,董洪亮,倪娜,等.PU.1和IL-9在肺纤维化大鼠周围免疫器官内的表达及其作用研究[J].免疫学杂志,2017,33(2):93-100.
[23]Li J,Chen S,Xiao X,et al.IL-9 and Th9 cells in health and diseases--From tolerance to immunopathology[J].Cytokine Growth Factor Rev,2017,37:47-55.
[24]陈杰,管俊昌.Th17/Treg细胞的免疫失衡与病原体的感染[J].蚌埠医学院学报,2018,43(10):1305-1308.