CAJ | 학술논문

目的在日本血吸虫早期感染模型和SEA致敏模型中,研究IGTP和IRG-47缺失小鼠在抵抗日本血吸虫感染过程中IL-10的免疫调节作用.方法以IGTP基因敲除(IGTP~(-/-))小鼠、IRG-47基因敲除(IRG-47~(-/-))小鼠和野生型(WT)小鼠为研究对象,建立日本血吸虫早期感染模型和可溶性虫卵抗原(SEA)致敏模型.在早期感染模型中,以HE染色观察耳廓的病理变化,间接ELISA法检测耳廓皮片培养上清中Th1/Th2型细胞因子的表达水平;在SEA致敏模型中,以间接ELISA法检测血清中日本血吸虫特异性IgG抗体、脾单个核细胞培养上清中Th1/Th2型细胞因子的表达水平;以及流式细胞术检测脾细胞亚群比例的变化.结果日本血吸虫感染7 d后,IGTP~(-/-)、IRG-47~(-/-)和WT小鼠的耳廓病理变化无显著差异;但IRG-47~(-/-)小鼠耳廓培养上清中IL-10水平显著低于IGTP~(-/-)小鼠.SEA致敏小鼠3周后,IGTP~(-/-)小鼠血清中SEA特异性IgG抗体水平显著低于WT小鼠;IRG-47~(-/-)小鼠脾细胞在体外SEA刺激下,产生的IL-10水平显著高于IGTP~(-/-)小鼠和WT小鼠;流式结果显示IRG-47~(-/-)小鼠脾中Th2细胞比例显著低于WT小鼠.结论 SEA是IRG-47缺失小鼠抵抗日本血吸虫、促使宿主产生保护性应答的重要调节因素,IL-10在此过程中是重要的免疫调节分子.
목적 재일본혈흡충조기감염모형화SEA치민모형중,연구IGTP화IRG-47결실소서재저항일본혈흡충감염과정중IL-10적면역조절작용.방법 이IGTP기인고제(IGTP~(-/-))소서、IRG-47기인고제(IRG-47~(-/-))소서화야생형(WT)소서위연구대상,건립일본혈흡충조기감염모형화가용성충란항원(SEA)치민모형.재조기감염모형중,이HE염색관찰이곽적병리변화,간접ELISA법검측이곽피편배양상청중Th1/Th2형세포인자적표체수평;재SEA치민모형중,이간접ELISA법검측혈청중일본혈흡충특이성IgG항체、비단개핵세포배양상청중Th1/Th2형세포인자적표체수평;이급류식세포술검측비세포아군비례적변화.결과 일본혈흡충감염7 d후,IGTP~(-/-)、IRG-47~(-/-)화WT소서적이곽병리변화무현저차이;단IRG-47~(-/-)소서이곽배양상청중IL-10수평현저저우IGTP~(-/-)소서.SEA치민소서3주후,IGTP~(-/-)소서혈청중SEA특이성IgG항체수평현저저우WT소서;IRG-47~(-/-)소서비세포재체외SEA자격하,산생적IL-10수평현저고우IGTP~(-/-)소서화WT소서;류식결과현시IRG-47~(-/-)소서비중Th2세포비례현저저우WT소서.결론 SEA시IRG-47결실소서저항일본혈흡충、촉사숙주산생보호성응답적중요조절인소,IL-10재차과정중시중요적면역조절분자.
Objective To investigate the immune response of IL-10 to Schistosoma japonicum infection in the early infectioin model and SEA immunization model of the IGTP~(-/-) and IRG-47~(-/-) mice.Methods In the early infection model,the IGTP knock out (IGTP~(-/-)) mice,IRG-47 knock out (IRG-47~(-/-)) mice and wild-type (WT) C57BL/6J mice were exposed to 300 S.japonicum cercariae via the pinna and sacrificed on day 7 post-infection.Each mouse pinna section was stained with hematoxylin-eosin (HE) to detect the pathological lesions,and the culture supernatant of pinna was used to test the levels of Th1/Th2 cytokines by indirect ELISA.In the SEA immunization model,IGTP~(-/-) IRG-47~(-/-) and WT mice were immunized with SEA twice and sacrificed in 3 weeks after the initial immunization.SEA-specific IgG antibody in sera was detected by indirect ELISA;the levels of Th1/Th2 cytokines were tested in culture supernatant of splenocytes by indirect ELISA;the proportions of CD4~+ T cells,CD8~+ T cells,B cells,Th1 and Th2 cells in the spleen were assayed by FACS.Results Although no obvious differences on the pathology of pinna were observed among the three mouse groups,the level of IL-10 in the culture supernatant of pinna of IRG-47~(-/-) mice was lower than that of IGTP~(-/-) mice in 7 days after the exposure.Following SEA immunization,the level of SEA-specific IgG antibody in sera of IGTP~(-/-) mice was lower than that in WT mice,the level of IL-10 in the culture supernatant of splenocytes of IRG-47~(-/-) mice was higher than that of IGTP~(-/-) and WT mice with the stimulation of SEA.However,the proportion of Th2 cells in the spleen of IRG47~(-/-) mice was the lowest among the three mouse groups.Conclusions SEA is the stimulus of IRG-47 deficiency mice to defend Schistosoma japanicum infection and promote the host to produce a protective response,and IL-10 may play an important role in immune regulation in this process.