摘要
目的：研究重组创伤弧菌溶细胞素（ rVvhA）对小鼠脾脏和肺脏T细胞亚群细胞毒活性作用及其组织差异性，了解其对脾脏和肺脏CD4^＋初始T细胞和效应T细胞存活率的影响。方法利用大肠杆菌表达系统重组表达创伤弧菌溶细胞素（rVvhA），体外作用于C57BL/6小鼠的脾脏和肺脏单细胞悬液，采用流式细胞术结合CD45.2、CD4、CD8、CD44、CD62L和Live/Dead等荧光抗体（或探针）检测分析rVvhA对CD4^＋T细胞和CD8^＋T细胞的存活率及组织差异性的影响。结果 rVvhA体外作用于脾脏和肺脏的单细胞悬液6 h后，小鼠脾脏CD4^＋T细胞和CD8^＋T细胞的存活率下降且CD4^＋T细胞对rVvhA的细胞毒性更为敏感，小鼠肺脏CD4^＋T细胞存活率轻微下降而CD8^＋T细胞的存活率无明显变化，此外，脾脏CD4^＋初始T细胞和效应T细胞的存活率均下降，而肺脏CD4^＋初始T细胞和效应T细胞的存活率均未有明显下降。结论 rVvhA对脾脏和肺脏T细胞亚群细胞毒活性存在组织差异性，而这种差异性可能与组织间的CD4^＋初始T细胞有关。
Objective To understand how Vibrio vulnificus hemolysin （ VvhA） affects the viability and tissue variability of murine spleen and lung T cell subsets. Methods The single-cell suspensions were prepared from spleen and lung tissues of C57BL/6 mice, and then were treated with recombinant VvhA （ rVvhA） for 6 hours in vitro. The viability and tissue variability of murine spleen and lung CD4^＋T and CD8^＋T cells were determined by staining with anti-mouse CD45. 2, CD4, CD8, CD44, CD62L and cell viability fluorescent dye or fluorescent antibody. Results After treated with the rVvhA for 6 hours in vitro, the via-bility of murine spleen CD4^＋T and CD8^＋T cells were decreased and the CD4^＋T cells were more sensitive to the cytotoxicity of rVvhA. However, there was no significant change in the viability of murine lung CD4^＋T and CD8^＋T cells. Furthermore, the viability of murine spleen na？ve CD4^＋T cells and effector CD4^＋T cells were also decreased, while there was no change in that of the na？ve CD4^＋T cells and effector CD4^＋T cells in lung tissues. Conclusion There was tissue variability in the cytotoxicity of rVvhA to murine spleen and lung T cell subsets and that might be associated with the na？ve CD4^＋T cells.
出处
《中华微生物学和免疫学杂志》
CAS
CSCD
北大核心
2016年第11期812-816,共5页
Chinese Journal of Microbiology and Immunology
