体外小鼠nuocyte细胞中IL-13的干预促进CD4^+T细胞向CD4^+IL10^+细胞的分化  

Intervention of IL-13 in nuocytes promotes differentiation of CD4^+IL10^+ cells from CD4^+T cells in vitro

作　　者：蔺林 陈峥 戴飞 魏瑾瑾 汤欣玥 LIN Lin;CHEN Zheng;DAI Fei;WEI Jin-jin;TANG Xin-yue(Department of Otorhinolaryngology-Head and Neck Surgery,Huashan Hospital North,Fudan University,Shanghai 201907,China)

机构地区：[1]复旦大学附属华山医院北院耳鼻咽喉头颈外科

出　　处：《复旦学报（医学版）》2019年第3期324-330,337,共8页Fudan University Journal of Medical Sciences

基　　金：国家自然科学基金面上项目(81371076);上海申康医院发展中心郊区医院临床能力建设项目(SHDC12015905);复旦大学附属华山医院北院启动基金(HSBY2016005)~~

摘　　要：目的探讨体外小鼠nuocyte细胞中IL-13的干预对CD4^+ IL10 +细胞的分化的影响。方法用卵清蛋白(ovalbumin,OVA)建立小鼠变应性鼻炎(allergic rhinitis,AR)模型( n =6),正常组注射生理盐水( n =6)。计数AR小鼠的打喷嚏、挠鼻次数,分离、提纯并体外培养小鼠鼻相关淋巴组织(nasal-associated lymphoid tissue,NALT)中的nuocyte细胞;构建针对小鼠IL-13的shRNA,并用慢病毒将IL-13 shRNA转染体外培养的AR小鼠nuocyte细胞,分别检测转染前后培养基中nuocyte细胞在鼠重组(recombinant,rm)IL-33作用下分泌IL-13和IL-10的浓度;获取AR小鼠外周血单个核细胞(peripheral blood mononuclear cells,PBMCs)并分离出CD4^+ T细胞进行体外培养,然后将体外培养的nuocyte细胞加入上述小鼠CD4^+ T细胞培养基共培养,检测CD4^+ IL10 +细胞所占CD4^+ T细胞的百分比。结果小鼠AR模型打喷嚏和挠鼻次数较正常小鼠显著增多,OVA诱导NALT来源的小鼠nuocyte细胞被鉴定为CD3CD4CD8CD19CD11bCD11cFcεR1(lineage)- ICOS +,AR小鼠的nuocyte细胞数较正常小鼠显著增多,携带有IL-13 shRNA的慢病毒成功转染到体外培养的nuocyte细胞。转染前经rmIL-33的刺激,培养基中IL-13和IL-10的浓度均升高,差异具有统计学意义;转染后培养基中IL-13的含量无明显增多,而IL-10含量明显增多,转染前后IL-13和IL-10的浓度差异具有统计学意义,IL-13浓度降低,IL-10浓度升高,而且转染后CD4^+ IL10 +细胞占CD4^+ T细胞的百分比也增加,差异具有统计学意义。结论体外小鼠nuocyte细胞中IL-13的干预可能促进CD4^+ IL10 +细胞的分化。Objective To investigate the influence of intervention of IL-13 in nuocytes on differentiation of CD4^+ IL10 + cells in vitro . Methods Mice allergic rhinitis (AR) models ( n =6) were established using ovalbumin (OVA),mice of control were injected with normal saline ( n =6).Numbers of sneezing and nasal rubbing were counted. Nuocytes were sorted and purified from the mouse nasal-associated lymphoid tissue (NALT),and the cells from AR mice NALT were cultured in vitro .Lentivirus vectors that encoded small hairpin RNA (shRNA) against IL-13 gene were established and administered into nuocytes cultures.Then,IL-13 and IL-10 were examined before and after transfection of nuocytes in response to recombinant (rm) IL-33. After that, mice peripheral blood mononuclear cells (PBMCs) were obtained,and CD4^+ T cells were sorted and purified from PBMCs,and cultured in vitro .Finally,NALT-derived nuocytes were co-cultured with CD4^+ T cells,and the percentage of CD4^+ IL10 + cells in CD4^+ T cells was evaluated. Results Numbers of sneezing and nasal rubbing were all increased in mice AR models compared with normal mice.OVA induced NALT-derived nuocytes were identified as CD3CD4CD8CD19CD11bCD11cFcεR1 (lineage)- ICOS +.Nuocyte number was increased in AR mice compared with normal ones.Lentivirus vectors that carried shRNA against IL-13 gene were established successfully,and transfected into cultured nuocytes in vitro .In response to rmIL-33,concentrations of IL-13 and IL-10 were all increased statistically before transfection.After transfection,IL-13 was not increased statistically,while IL-10 was elevated significantly after transfection.The concentrations of IL-13 and IL-10 before and after transfection were statistically changed.IL-13 concentration was decreased while IL-10 concentrations was increased.Furthermore,the percentage of CD4^+ IL10 + cells was also increased statistically after nuocytes transfection. Conclusions Intervention of IL-13 in nuocytes might promote differentiation of CD4^+ IL10 + cells from CD4^+ T cells i

关 键 词：nuocyte细胞 CD4+IL10+细胞 变应性鼻炎(AR) 鼻相关淋巴组织(NALT) 白介素(IL) 小鼠 

分 类 号：R765[医药卫生—耳鼻咽喉科]