脂多糖通过调控树突状细胞存活和分泌细胞因子促进CD4^+T细胞增殖  

Lipopolysaccharide promotes the proliferation of CD4^+ T cells by modulating the survive and cytokine secretion of dendritic cells

作　　者：陈松 卢利莎 王伟强[2] 薛婷[3] 余娟[3] 孙志娜 赵春晓 廖芳[3] 

机构地区：[1]中国医学科学院血液病医院血液学国家重点实验室,天津300020 [2]天津医科大学总医院消化科,天津300052 [3]华中科技大学同济医学院基础医学院微生物教研室,湖北武汉430030

出　　处：《细胞与分子免疫学杂志》2016年第9期1153-1157,1163,共6页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家重大科学研究计划(2013CB966904;2015CB964402);国家自然科学基金(81421002;81273217;81322007)

摘　　要：目的探索脂多糖(LPS)调节树突状细胞(DC)的功能,促进T细胞免疫反应的机制。方法应用CDllc^+免疫磁珠分离小鼠脾脏DC。LPS处理DC后。流式细胞术检测DC表面的共刺激分子CD80和CD86的表达,ELISA检测DC培养上清中白细胞介素4(IL-4)、IL-5、IL-6、IL-12p40、IL-12p70、肿瘤坏死因子α(TNF-α)的水平,异硫氰酸荧光素标记的膜联素Ⅴ/碘化丙啶(annexinⅤ-FITC/PI)双染色结合流式细胞术检测DC凋亡,Phos-flow技术检测核因子κB P65(NF-κB P65)磷酸化水平,实时定量PCR检测基因芯片变化差异明显基因的mRNA水平,DC经OVA323-329多肽处理后与和CD4^+T细胞共培养,流式细胞术检测CD4^+T细胞的增殖。结果利用CD11 c磁珠分离,可获得纯度为93%的DC,LPS可以上调DC表面CD80和CD86的表达,增强DC介导的CD4^+T细胞的增殖。并且LPS能促进促炎细胞因子IL-12 p40、TNF-α和IL-6的分泌,同时通过NF-κB通路抑制DC的凋亡。结论 LPS通过调节DC的存活和细胞因子的产生促进DC介导的CD4^+T细胞的增殖。Objective To explore the mechanism modulating the function of dendritic cells （DCs） and promoting the T cell response by lipopolysaccharide （LPS）. Methods Splenic DCs were purified with anti-CDllc immunomagnetic beads. After DCs were stimulated with LPS, the expressions of co-stimulatory molecules CD80 and CD86 on the DCs were detected by flow cytometry. The protein levels of pre-inflamrnatory cytokines interleukin 4 （IL-4）, IL-5, IL-6, IL-12p40, IL-12p70 and tumor necrosis factor alpha （TNF-α） in the culture supernatant were measured by ELISA. The apoptotic levels of DCs which were labeled with annexinV-FITC/PI were determined by flow cytometry. The phosphorylation level of nuclear factor κB P65 （NF-κB P65） was assessed by phos-flow. The mRNA levels of variable genes in microarray were determined by real-time PCR. The proliferation of CD4＋ T cells which were co-cultured with OVA323-329-treated DCs was analyzed by flow cytometry. Results The purity of DCs reached over 93% after isolation. LPS up-regulated the expressions of CDS0 and CD86 and enhanced DCs-mediated proliferation of CD4＋ T cells. In addition, LPS increased the protein levels of IL-12p40, TNF-α and IL-6, and inhibited the apoptosis of DCs through the NF-κB signaling pathway. Conclusion LPS could enhance DC-mediated proliferation of CD4＋ T cells by modulating the DCs survival and cytokine secretion.

关 键 词：树突状细胞 脂多糖 凋亡 CD4+T细胞 

分 类 号：R392.12[医药卫生—免疫学]