rhHMGB1对HTLV-1＋MT2细胞中HMGB1相关受体表达的影响  

Influences of recombinant human HMGB1 on the expression of HMGB1-related receptors in HTLV-1 infected MT2 cells

作　　者：李向平[1,2] 牛志国[1] 韩静贤[1] 高彩[1] 刘威[1] 宋向凤[1] 高志涛[1] 王辉[1] 

机构地区：[1]新乡医学院免疫学研究中心,453003 [2]三全学院

出　　处：《中华微生物学和免疫学杂志》2014年第1期47-50,共4页Chinese Journal of Microbiology and Immunology

基　　金：国家自然科学基金（30972755,81273241）;河南省教育厅科学技术研究重点项目（13A310843）;新乡医学院重点领域招标课题（ZD2011-15）;新乡医学院研究生创新课题资助计划（YJSCX201201Z）

摘　　要：目的探讨重组人高迁移率族蛋白1（recombinant human HMGB1，rhHMGB1）对人T淋巴细胞白血病病毒1型（HTLV-1）感染的T细胞中与HMGB1相关膜受体表达的影响。方法病毒阳性细胞MT2中加入终浓度为1μg／ml的人源化重组人rhHMGB1蛋白或等体积的PBS，培养24h后，以流式细胞术、免疫印迹、免疫荧光检测膜受体晚期糖基化终产物受体（receptor for advanced glycosylation end products，RAGE）、T细胞相关免疫球蛋白黏蛋白3（T cell immunoglobulin mucin3，TIM-3）、Toll样受体4（Toll like receptor4，TLR4）的表达及分布。结果流式细胞术检测结果显示1μg／ml rhHMGB1刺激MT2细胞后，与对照相比RAGE表达下降，TLR4的表达明显升高（P〈0．05）；免疫印迹检测结果显示1μg／ml rhHMGB1刺激MT2细胞后，细胞总蛋白和胞质中，膜受体RAGE和TLR4表达均明显升高（P〈0．05），而TIM-3的表达没有明显变化；免疫荧光检测结果显示rhHMGB1刺激MT2细胞后TLR4和RAGE的荧光强度均有增加，而TIM-3的荧光强度没有明显变化，且TLR4、TIM-3集中于细胞膜，而RAGE则集中于细胞质，细胞膜表达较少。结论胞外的HMGB1可促进HTLV-1感染T细胞RAGE和TLR4的表达。Objective To investigate the influences of recombinant human high mobility group protein 1 （ rhHMGB1 ） on the expression of HMGB1-related receptors in Human T-cell leukemia virus type 1 （ HTLV-1 ） infected MT2 T cells. Methods rhHMGB1 was added to HTLV-l-positive MT2 cells at a final concentration of 1 μg/ml. Equal volume of PBS was used to set up the control. After 24 h of culture, the expression and distribu- tion of surface molecules including advanced glycosylation end products （RAGE）, T cell immunoglobulin mucin 3 （TIM-3） and toll like receptor 4 （TLR4） were detected by flow cytometry, Western blot and immunofluores- cence. Results The results of flow cytometry analysis showed a decreased expression of RAGE, but a signifi- cantly increased expression of TLR4 （P〈0.05） on MT2 cells stimulated by 1μg/ml rhHMGB1, as compared with control group. The expression of RAGE and TLR4 among total cellular proteins and cytoplasmic proteins were significantly increased （ P〈0.05）, while the expression of TIM-3 showed no significant change as indicated by Western Blot. The immunofluorescence staining demonstrated that the fluorescence intensity of TLR4 and RAGE were increased, while the fluorescence intensity of TIM-3 showed no change upon rhHMGB1 treatment. Furthermore, TLR4 and TIM-3 were mainly distributed on the cell membrane, while RAGE was found in the cy- toplasm. Conclusion Recombinant human HMGB1 can enhance the expression of RAGE and TLR4 in HTLV- 1-infected T cell line MT2.

关 键 词：成人T淋巴细胞白血病病毒1型 高迁移率组蛋白1 HMGB1相关膜受体 

分 类 号：R363[医药卫生—病理学]