全葡聚糖颗粒促进肿瘤微环境驯化的小鼠树突状细胞成熟并抑制调节性T细胞分化  

Whole glucan particle promotes the maturation of tumor-educated mouse dendritic cells to suppress regulatory T cell differentiation

作　　者：白煜 宁永玲[2] 周洪[1] 戚春建[2] 

机构地区：[1]南京医科大学基础医学院,江苏南京210029 [2]南京医科大学附属常州市第二人民医院中心实验室,江苏常州213164

出　　处：《细胞与分子免疫学杂志》2017年第9期1171-1176,共6页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金(81672799,31500731);常州市高层次卫生人才培养工程(2016CZLJ016,2016CZBJ013)

摘　　要：目的体外模拟肿瘤微环境驯化免疫抑制性树突状细胞(TEDC),研究全葡聚糖颗粒(WGP)对其免疫功能的影响。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),同时加入白细胞介素10(IL-10)、转化生长因子β(TGF-β)共培养,诱导得到TEDC,再加入WGP激发2 d。磁珠分选出OT-Ⅱ小鼠淋巴结和脾脏中的CD4+T细胞,加入特异性抗原卵清蛋白(OVA),再与TEDC共培养。流式细胞术检测细胞的CD11c、CD86、CD40、CD80、MHC-Ⅱ的水平和CD4+FOXP3+调节性T细胞(Treg)的分化情况。实时荧光定量PCR检测TEDC的肿瘤坏死因子α(TNF-α)、IL-12p40、IL-10、IL-6、IL-4、TGF-β1 m RNA水平;ELISA检测TEDC培养上清中的TNF-α、IL-23、IL-12p70、IL-4水平。结果WGP可以上调TEDC表面MHC-Ⅱ、CD86、CD80和CD40的表达,提高TNF-α、IL-12p40、IL-6水平,降低TGF-β1水平;抑制其诱导CD4+FOXP3+T细胞分化能力。结论β-葡聚糖可以改变TEDC的成熟和分化状态,改善其免疫抑制功能。Objective To investigate the maturation and immune responses of tumor-educated dendritic cells（ TEDCs）stimulated by whole glucan particle（ WGP）. Methods Mouse bone marrow-derived dendritic cells were generated by Flt3 ligand and co-cultured with interleukin 10（ IL-10） and transforming growth factor β（ TGF-β） in vitro to induce TEDCs,and then stimulated by WGP for 2 days. CD4＋FOXP3＋T cells from the lymph nodes and spleens of OT-Ⅱ mice were purified by magnetic-activated cell sorting,and then co-cultured with TEDCs in the presence of ovalbumin（ OVA）. Expressions of cell surface markers（ CD11 c,CD86,CD40,CD80,MHC-Ⅱ） and CD4＋FOXP3＋T cell differentiation and proliferation were detected by flow cytometry. real-time quantitative PCR was used to detect the m RNA levels of TNF-α,IL-12 p40,IL-10,IL-6,IL-4,TGF-β1 and ELISA was used to detect the levels of TNF-α,IL-23,IL-12 p70,IL-4 in culture supernatant. Results WGP up-regulated MHC-Ⅱ and co-stimulation molecules CD86,CD80,CD40 on TEDCs,increased the levels of TNF-α,IL-12 p40,IL-6 in TEDCs,decreased the production of TGF-β1. Furthermore,WGP also suppressed CD4＋FOXP3＋T cell differentiation and proliferation in vitro. Conclusion WGP improves TEDC maturation and inhibits immunosuppressive functions.

关 键 词：全葡聚糖颗粒(WGP) 树突状细胞(DC) 调节性T细胞(Treg) 

分 类 号：R392.1[医药卫生—免疫学] R730.3[医药卫生—基础医学]