CAJ | 학술논문

目的：观察足细胞树突状细胞（DC）表型分子DC?SIGN的表达，以及探讨其对狼疮肾炎（LN）局部免疫炎性反应的作用。方法利用免疫组化及免疫荧光观察LN患者肾组织DC?SIGN及IgG1表达。选取4周龄LN小鼠随机分为实验组、干预组，另取C57BL/6J小鼠作为正常对照组。干预组于小鼠第6周龄注入抗DC?SIGN抗体，分别于第16、20、24、28周龄处死各组小鼠，观察小鼠肾功能及肾组织病理变化，并采用免疫组化及免疫荧光观察肾组织DC?SIGN及IgG1表达。此外取小鼠足细胞给予LN鼠血清处理，流式细胞术观察足细胞主要组织相容性复合体II（MHC II）、CD80、DC?SIGN表达；混合淋巴细胞反应（MLR）检测共培养后其刺激T细胞增殖的能力；ELISA法测定MLR上清中γ干扰素（IFN?γ）、IL?4含量。结果（1）LN患者肾小球可见DC?SIGN及IgG1表达。（2）LN小鼠自20周龄起伴随尿蛋白增加（P＜0.01），肾小球亦见DC?SIGN及IgG1表达，至24周龄Scr显著增加（P＜0.01）；经针对DC?SIGN抗体干预后，尿蛋白减少（P＜0.01），肾功能明显改善（P＜0.01）。（3）经LN鼠血清刺激后，小鼠足细胞DC?SIGN、MHC II、CD80表达上调，体外刺激T细胞增殖能力增强（P＜0.01），且T细胞分泌的IFN?γ/IL?4比值增高（P＜0.01）；经抗DC?SIGN抗体干预，足细胞DC?SIGN、MHC II、CD80表达均下调，且刺激T细胞增殖能力减弱（P＜0.01），IFN?γ/IL?4比值下降（P＜0.01）。结论 LN足细胞可通过表达DC?SIGN发挥DC样细胞功能，可能参与LN局部组织免疫炎性反应；而抑制DC?SIGN可调抑足细胞免疫功能，并产生防治效应。
목적：관찰족세포수돌상세포（DC）표형분자DC?SIGN적표체，이급탐토기대랑창신염（LN）국부면역염성반응적작용。방법이용면역조화급면역형광관찰LN환자신조직DC?SIGN급IgG1표체。선취4주령LN소서수궤분위실험조、간예조，령취C57BL/6J소서작위정상대조조。간예조우소서제6주령주입항DC?SIGN항체，분별우제16、20、24、28주령처사각조소서，관찰소서신공능급신조직병리변화，병채용면역조화급면역형광관찰신조직DC?SIGN급IgG1표체。차외취소서족세포급여LN서혈청처리，류식세포술관찰족세포주요조직상용성복합체II（MHC II）、CD80、DC?SIGN표체；혼합림파세포반응（MLR）검측공배양후기자격T세포증식적능력；ELISA법측정MLR상청중γ간우소（IFN?γ）、IL?4함량。결과（1）LN환자신소구가견DC?SIGN급IgG1표체。（2）LN소서자20주령기반수뇨단백증가（P＜0.01），신소구역견DC?SIGN급IgG1표체，지24주령Scr현저증가（P＜0.01）；경침대DC?SIGN항체간예후，뇨단백감소（P＜0.01），신공능명현개선（P＜0.01）。（3）경LN서혈청자격후，소서족세포DC?SIGN、MHC II、CD80표체상조，체외자격T세포증식능력증강（P＜0.01），차T세포분비적IFN?γ/IL?4비치증고（P＜0.01）；경항DC?SIGN항체간예，족세포DC?SIGN、MHC II、CD80표체균하조，차자격T세포증식능력감약（P＜0.01），IFN?γ/IL?4비치하강（P＜0.01）。결론 LN족세포가통과표체DC?SIGN발휘DC양세포공능，가능삼여LN국부조직면역염성반응；이억제DC?SIGN가조억족세포면역공능，병산생방치효응。
Objective To explore the expression of DC?SIGN, the phenotype of dendritic cells (DCs), on podocytes, and its role in immune and inflammatory responses of lupus nephritis (LN). Methods DC?SIGN and IgG1 expression in renal tissues of lupus nephritis patients were observed by immunohistochemistry and immunofluorescence. The 4?week old LN mice were randomly divided into the experimental group and the intervention group. C57BL/6J mice were used as normal control group. Mice of the intervention group were injected anti?DC?SIGN antibody at 6?week old. Mice were sacrificed at 16, 20, 24, 28?week old respectively, to observe the mice renal function and pathological changes. And DC?SIGN and IgG1 expression in renal tissue were observed by immunohistochemistry and immunofluorescence. In addition, mice podocytes were treated with serum of LN mice. Flow cytometry was used to investigate the expression of MHC II, CD80 and DC?SIGN expression on podocytes. Mixed lymphocyte reaction was used to detect the ability of stimulating T cells proliferation. IFN?gamma and IL?4 in supernatant were determined by ELISA. Results (1) Expression of DC?SIGN and IgG1 was found in glomeruli of lupus nephritis patients. (2) Accompanied by increased proteinuria of LN mice from 20?week old (P<0.01), DC?SIGN and IgG1 expression was found in glomeruli, and the renal function deteriorated up to 24 week?old (P<0.01). Mice with anti?DC?SIGN antibody intervention appeared reduced proteinuria and remission of renal function (P<0.01). (3) After stimulated by serum of LN mice, the expression of DC?SIGN, MHC II and CD80 was up?regulated, stimulation of T cell proliferation was enhanced (P<0.01), and IFN?gamma/IL?4 ratio increased (P<0.01). Anti?DC?SIGN antibody treatment down?regulated the expressions of DC?SIGN, MHC II and CD80 on podocytes, decreased the ability of stimulating T cell proliferation and lowered the ratio of IFN?gamma/IL?4 (P<0.01). Conclusions Podocytes in lupus nephritis can play DC?like function through the expression of DC?SIGN, which may be involved in immune and inflammatory responses of renal tissue. However, inhibiton of DC?SIGN can depress immune function of podocytes and have prevention and treatment effect.