Neuropilin1+T细胞及CD4+CD25+调节性T细胞比较.docVIP

Neuropilin1+T细胞及CD4+CD25+调节性T细胞比较 　 作者：裴向克, 蔡明, 周鸿敏, 刘畅, 徐利军, 陈忠华, 石炳毅 【摘要】 　　目的: 分析Neuropilin1+T细胞(Nrp1+T细胞)与经典CD4+CD25+调节性T细胞(Treg)的关系并比较二者的免疫调节作用。方法: 流式细胞术分析BALB/c小鼠脾脏T细胞上Nrp1与CD4、 CD25的表达关系并分选Nrp1+T细胞及CD4+CD25+Treg, 通过B16F10lucG5黑色素肿瘤细胞体外培养实验并利用萤光成像系统, 观察比较两种细胞对NK细胞杀伤B16F10lucG5黑色素瘤细胞的影响。结果: CD4+CD25+Treg中表达Nrp1的比例为(27.28±1.17)%, 明显高于CD4+CD25-T细胞的(1.63±0.08)% (P＜0.01); 在体外实验中, Nrp1+T细胞与CD4+CD25+Treg均能抑制NK细胞杀伤B16F10lucG5黑色素瘤细胞, Nrp1+T细胞组的肿瘤细胞数目在6、 24、 48、 72 h分别为984±15、 1015±14、 1261±21、 1323±38, 高于CD4+CD25+Treg组的931±4、 983±8、 1201±18、 1256±18, 两组肿瘤细胞数目在各时间点均有统计学意义(P＜0.01)。结论: 经典CD4+CD25+Treg中表达Nrp1的细胞比例较高, Nrp1+T细胞有负性免疫调节作用, 抑制功能比CD4+CD25+Treg更强, 可以作为一类新的Treg亚群。 【关键词】 Neuropilin1; 调节性T细胞; 萤光成像 　　［Abstract］ AIM: To analyze the relationship between Nrp1(neuropilin1) positive T cells (Nrp1+T cells) and CD4+CD25+ regulatory T cells (CD4+CD25+Treg) and compare their immunoregulatory effects. METHODS: The expression of Nrp1, CD4 and CD25 on the splenic T cells of BALB/c mice was detected by flow cytometry and Nrp1+T cells and CD4+CD25+Treg were sorted. Then their effects of on NK cells killing B16F10lucG5 melanoma cells were compared in vitro by bioluminescence imaging system. RESULTS: The expression of Nrp1 on CD4+CD25+Treg was (27.28±1.17)%, which was significantly higher than that (1.63±0.08)% on CD4+CD25- T cells(P＜0.01). Both Nrp1+T cells and CD4+CD25+Treg inhibited the effect of killing NK cells on B16F10lucG5 melanoma cells in vitro. At 6, 24, 48 and 72 h, the number of tumor cells in Nrp1+T cell group was higher than that in CD4+CD25+Treg group(984±15 vs 931±4, 1015±14 vs 983±8, 1261±21 vs 1201±18 and 1323±38 vs 1256±18, respectively). There was significant statistical difference between the two groups at each time point (P＜0.01). CONCLUSION: The proportion of CD4+CD25+Treg expressing Nrp1 is high. Nrp1+T cells show negtive immunoregulatory effect, which is more powerful than CD4+CD25+Treg. Nrp1+T cells may be reg