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目的:比较正常人周围血中CD3+TCRvβ11+NKT细胞和CD3+TCRvα24+NKT细胞在频率、亚群、表型特征及功能方面的异同,以进一步了解NKT细胞在免疫应答中的作用。方法:分离正常成年人PBMCs,利用流式细胞术(FCM)检测TCRvα24、TCRvβ11、CD4、CD8、CD45RA、CD62L和CCR7表面分子的表达;PMA+Ionomyc in刺激PBMCs后,检测CD3+TCRvα24+、CD3+TCRvβ11+NKT细胞产生细胞因子IL-4和IFN-γ的情况。结果:PBMCs中CD3+TCRvα24+和CD3+TCRvβ11+NKT细胞的平均频率分别为0.63%和0.43%,NKT细胞频率的个体差异较大,少数细胞同时表达TCRvα24和TCRvβ11;根据CD4和CD8分子的表达,PBMCs中CD3+TCRvα24+NKT细胞可分为CD4+、CD8+、CD4-CD8-3个亚群,平均频率分别为64.35%、19.04%、17.18%,CD3+TCRvβ11+NKT细胞同样可分为CD4+、CD8+、CD4-CD8-3个亚群,其平均频率分别为53.69%、18.99%、29.74%,相应各亚群之间无显著差异;CD45RA+CD3+TCRvβ11+NKT细胞的频率(71.14%)要高于CD45RA+CD3+TCRvα24+NKT细胞的频率(46.55%),二者之间差异有显著性,CD62L+CD3+TCRvα24+NKT(46.26%)对CD62L+CD3+TCRvβ11+NKT(42.36%)以及CCR7+CD3+TCRvα24+NKT(9.24%)对CCR7+CD3+TCRvβ11+NKT(8.22%)之间的差异均无统计学意义;细胞因子检测的结果表明CD3+TCRvα24+NKT细胞和CD3+TCRvβ11+NKT细胞产生的IL-4(13.01%对6.62%)和IFN-γ(38.12%对26.95%)的总体水平间无显著性差异,但是IFN-γ+IL-4+CD3+TCRvα24+NKT细胞的平均频率(12.65%)要高于IFN-γ+IL-4+CD3+TCRvβ11+NKT细胞的平均频率(3.02%),且二者之间的差异有统计学意义。结论:正常人周围血中CD3+TCRvα24+NKT细胞和CD3+TCRvβ11+NKT细胞在频率、表型及产生细胞因子方面均有一定差异,总体来看,二者频率虽小但表型复杂,产生细胞因子IFN-γ和IL-4的水平高,参与免疫调节及免疫应答的过程。
OBJECTIVE: To compare the similarities and differences in frequency, subpopulation, phenotypic characteristics and function of CD3 + TCRvβ11 + NKT cells and CD3 + TCRvα24 + NKT cells in normal human peripheral blood in order to further understand the role of NKT cells in immune response. Methods: PBMCs from normal adults were isolated and the expression of TCRvα24, TCRvβ11, CD4, CD8, CD45RA, CD62L and CCR7 were detected by flow cytometry (FCM). After PBMCs were stimulated with PMA + Ionomyc in, the expressions of CD3 + TCRvα24 +, CD3 + TCRvβ11 + NKT cells produce cytokines IL-4 and IFN-γ. Results: The average frequencies of CD3 + TCRvα24 + and CD3 + TCRvβ11 + NKT cells in PBMCs were 0.63% and 0.43%, respectively. The frequency of NKT cells was significantly different between individuals. A few cells expressed TCRvα24 and TCRvβ11 simultaneously. According to the expression of CD4 and CD8, The average frequency of CD3 + TCRvα24 + NKT cells in PBMCs was 64.35%, 19.04% and 17.18%, respectively. The CD3 + TCRvα11 + NKT cells were also divided into CD4 +, CD8 + and CD4- , CD4-CD8-3 subsets, the average frequencies were 53.69%, 18.99% and 29.74%, respectively, with no significant difference among the corresponding subpopulations. The frequency of CD45RA + CD3 + TCRvβ11 + NKT cells was 71.14% (46.55%) in CD45RA + CD3 + TCRvα24 + NKT cells, the difference was significant between CD62L + CD3 + TCRvα24 + NKT (46.26%) and CD62L + CD3 + TCRvβ11 + NKT CD3 + TCRvα24 + NKT (9.24%) were not significantly different from those of CCR7 + CD3 + TCRvβ11 + NKT (8.22%). The results of cytokine test showed that CD3 + TCRvα24 + NKT cells and CD3 + TCRvβ11 + NKT There was no significant difference in the overall level of IL-4 (13.01% vs 6.62%) and IFN-γ (38.12% vs 26.95%) produced by the cells but the mean IFN- [gamma] + IL-4 + CD3 + TCRv [alpha] 24 + NKT cells frequency (12.65%) than IFN-γ + IL-4 + CD3 + TCRvβ11 + average frequency of NKT cells (3.02%), there were statistically significant differences between the two. Conclusion: The frequency, phenotype and cytokine production of CD3 + TCRvα24 + NKT cells and CD3 + TCRvβ11 + NKT cells in peripheral blood of normal people are all different. Generally speaking, the frequency of CD3 + Cytokines IFN-γ and IL-4 levels are high, involved in immune regulation and immune response process.