经HIV包膜蛋白gp120刺激的T细胞外泌体促进巨噬细胞M2极化  

Extracellular vesicles from T cells stimulated by HIV envelope protein gp120 promote macrophage M2 polarization

作　　者：韦武均 黄晶晶 覃林秀 柴富 李嘉兴 林成 钟丽梅[5] 黎作茶[1] 胡仁统[1] 庞晓霞[1] 韦家住 陈晓昊 王春芳[1] WEI Wu-jun;HUANG Jing-jing;QIN Lin-xiu;CHAI Fu;LI Jia-xing;LIN Cheng;ZHONG Li-mei;LI Zuo-cha;HU Ren-tong;PANG Xiao-xia;WEI Jia-zhu;CHEN Xiao-hao;WANG Chun-fang(Department of Clinical Laboratory,Affiliated Hospital of Youjiang Medical University for Nationalities,Baise 533000,China;Department of Health Care,Baise Maternal and Child Health Hospital,Baise 533000,China;Department of Pharmacy,Affiliated Hospital of Youjiang Medical University for Nationalities,Baise 533000,China;Department of Oncology,Affiliated Hospital of Youjiang Medical University for Nationalities,Baise 533000,China;Department of Emergency Medicine,Affiliated Hospital of Youjiang Medical University for Nationalities,Baise 533000,China)

机构地区：[1]右江民族医学院附属医院医学检验科,广西百色533000 [2]百色市妇幼保健院保健科,广西百色533000 [3]右江民族医学院附属医院药剂科,广西百色533000 [4]右江民族医学院附属医院肿瘤科,广西百色533000 [5]右江民族医学院附属医院急诊科,广西百色533000

出　　处：《海南医学院学报》2023年第24期1841-1847,共7页Journal of Hainan Medical University

基　　金：国家自然科学基金项目(81960303);百色市科学研究与技术开发计划项目(百科20213301,百科20213242,百科20194701);广西卫生健康委员会自筹经费科研课题(Z20190202,Z20211114);广西高校中青年教师科研基础能力提升项目(2021KY0538);2022年中医药自筹经费科研课题(GXZYL20220304);广西肝胆疾病分子病理学重点实验室开放课题(GXZDSYS‑009)。

摘　　要：目的:本研究旨在探讨经HIV膜蛋白gp120处理后人T淋巴细胞系(H9)外泌体对巨噬细胞极化的影响。方法:采用CCK8试剂盒检测gp120处理后人T淋巴细胞H9活力;采用ELISA法检测H9细胞上清液中炎症因子水平;通过电镜和Western blot实验鉴定外泌体特征;PHK67染色观察外泌体进入巨噬细胞情况;最后通过ELISA和免疫荧光检测巨噬细胞极化标记物,分析gp120处理后T细胞外泌体对巨噬细胞极化的影响。结果:HIV gp120蛋白抑制人T淋巴细胞H9增殖并促进炎症因子释放。成功提取人T淋巴细胞H9外泌体,电镜下为中间凹陷的膜结构,高表达标记蛋白CD9、CD63、CD81。PHK67染色结果显示H9细胞外泌体可进入巨噬细胞。经gp120处理的H9细胞外泌体可促进巨噬细胞向M2型极化。结论:HIV膜蛋白gp120处理人T淋巴细胞H9分泌的外泌体能够促进巨噬细胞M2极化,可能是gp120在免疫调节中的新机制。Objective:This study aimed to investigate the effects of exosomes from human T lymphocyte line(H9)treated with HIV envelope protein gp120 on macrophage polarization.Methods:The viability of gp120-treated H9 T lymphocytes was assessed using the CCK8 assay.Inflammatory cytokine levels in the supernatant of H9 cells were determined by ELISA.Exosome characteristics were identified through electron microscopy and Western blot experiments.PHK67 staining was employed to observe the uptake of exosomes by macrophages.Finally,macrophage polarization markers were detected using ELISA and immunofluorescence to analyze the impact of gp120-treated T cell exosomes on macrophage polarization.Results:HIV gp120 protein inhibited the proliferation of human T lymphocytes H9 and promoted the release of inflammatory cytokines.Exosomes from H9 T lymphocytes were successfully isolated,displaying a cup-shaped membranous structure under electron microscopy and overexpressing marker proteins CD9,CD63,and CD81.PHK67 staining results indicated that exosomes from H9 cells could be internalized by macrophages.Exosomes from gp120-treated H9 cells promoted the polarization of macrophages towards the M2 phenotype.Conclusion:Exosomes secreted by human T lymphocytes H9 treated with HIV envelope protein gp120 can promote M2 polarization of macrophages,suggesting a potential novel mechanism of gp120 in immune modulation.

关 键 词：HIV GP120 外泌体 巨噬细胞 极化 

分 类 号：R392[医药卫生—免疫学]