全葡聚糖颗粒对DC诱导CD4^+T细胞向Th17细胞分化的影响  

The effect of whole glucan particle on CD4^+T cells differentiation toward Th17 cells induced by dendritic cells

作　　者：谢叶文 宁永玲[1] 潘洁 陈洁[1] 戚春建[1] XIE Yewen;NING Yongling;PAN Jie;CHEN Jie;QI Chunjian(Medical Research Center,Changzhou Second People’s Hospital Affiliated to Nanjing Medical University,Changzhou 213164,China)

机构地区：[1]南京医科大学附属常州市第二人民医院中心实验室,常州213164

出　　处：《免疫学杂志》2020年第6期504-509,共6页Immunological Journal

基　　金：国家科学基金(81672799);江苏省“333工程”科研项目(BRA2016121);江苏省科技计划重点项目(BE2019651)。

摘　　要：目的研究全葡聚糖颗粒(whole glucan particle,WGP)对DC诱导CD4^+T细胞向Th17细胞分化的影响。方法采用重组小鼠FMS样酪氨酸激酶3配体(Flt3L)法诱导C57BL/6小鼠骨髓来源树突状细胞(BMDC)。用MACS磁珠分选法对OT-Ⅱ小鼠的脾脏和淋巴结中CD4^+T细胞进行纯化,加入特异性抗原卵清蛋白(OVA),与DC共培养。实验分组为:CON组、WGP组、TGP-β组、WGP+TGF-β组,培养5 d后,利用流式细胞术(FCM)检测各组中CD4^+T细胞增殖和Th17细胞变化情况,ELISA检测各组细胞上清液中IL-17A的含量,Q-PCR检测各组细胞中Th17细胞核转录因子维甲酸孤儿受体-γt(retinoid-related orphan receptor-γt,ROR-γt)mRNA表达水平。结果TGF-β可以促进DC诱导CD4^+T细胞向Th17分化,而经WGP激发后,T细胞显著下调ROR-γt的转录水平,并减少对IL-17A的分泌。结论WGP显著抑制TGF-β诱导CD4+T细胞向Th17细胞分化。This study was performed to investigate the effect of whole glucan particle(WGP)on dendritic cells(DCs)-induced CD4^+T cells differentiation toward Th17 cells.Mouse bone marrow-derived dendritic cells were generated by Flt3 ligand.CD4^+T cells from the lymph nodes and spleens of OT-Ⅱmice were purified by magneticactivated cell sorting,and then co-cultured with DCs in the presence of ovalbumin(OVA).CD4^+T cells were divided into CON group,WGP group,TGF-βgroup and WGP+TGF-βgroup,and then cultured respectively for 5 days.The expression of cell surface marker IL-17 A and the CD4^+T cell proliferation were detected by flow cytometry(FCM);the cytokine of IL-17 in the supernatant were measured by ELISA.Real-time quantitative PCR was used to detect the mRNA level of ROR-γt.Data showed that TGF-βpromoted DCs to induce CD4^+T cells differentiation towards to IL-17^+T cell,meanwhile WGP significantly decreased ROR-γt transcription level and IL-17 A secretion of CD4^+T cells,resulting in suppressing Th17 cells differentiation.The results above suggest that WGP can inhibit TGF-βinduced differentiation of CD4^+T cells into Th17 cells.

关 键 词：全葡聚糖颗粒 DC细胞 转化生长因子-Β TH17细胞 

分 类 号：R392.12[医药卫生—免疫学]