外周血单个核细胞中CD3⁺TCRvα24⁺自然杀伤T细胞与CD3⁺TCRvβ11⁺自然杀伤T细胞的差异

[Differences between CD3⁺ TCRvα24⁺ NKT cell and CD3⁺ TCRvβ11⁺ NKT cell in PBMC].

作者信息

Li Zi-Tao, Yang Bin-Yan, Wu Chang-You

机构信息

Institute of Immunology, Zhongshan School of Medicine, State Ministry of Education Key Laboratory of Tropical Disease Control Research, Sun Yat-sen University, Guangzhou 510080, China.

出版信息

Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2011 Sep;27(9):949-53.

PMID:21906465

Abstract

AIM

Clarified the differences between CD3(+);TCRvα24(+); NKT cells and CD3(+);TCRvβ11(+); NKT cells in their frequencies, subpopulations, phenotypes and biological functions, so as to fully understand the effects of NKT cells in immune responses.

METHODS

PBMCs from blood donors were isolated and cell surface markers (CD3, TCRvα24, TCRvβ11, CD4, CD8, CD45RA, CD62L, CCR7) and intracellular cytokines (IL-4, IFN-γ) were detected by flow cytometry directly or after stimulation with PMA plus Ionomycin.

RESULTS

The mean frequencies of CD3(+);TCRvα24(+); NKT cells and CD3(+);TCRvβ11(+); NKT cells in PBMCs were 0.63% and 0.43% and they varied according to individuals. A small population of NKT cells coexpressed TCRvα24 and TCRvβ11. The subpopulations of CD4(+); NKT 64.35%, CD8(+); NKT 19.04%, CD4(-);CD8(-); NKT 17.18% in human CD3(+);TCRvα24(+); NKT cells and CD4(+); NKT 53.69%, CD8(+); NKT 18.99%, CD4(-);CD8(-); NKT 29.74% in CD3(+);TCRvβ11(+); NKT cells could be identified based upon the expressions of CD4 and CD8 molecules. There were no significant differences between relative subtypes. The frequency of CD45RA(+);CD3(+);TCRvβ11(+); NKT cells(71.14%) was higher than the frequency of CD45RA(+);CD3(+);TCRvα24(+); NKT cells and the differences between them were significant. The differences between the frequencies of CD62L(+);CD3(+);TCRvα24(+); NKT cells(46.26%) and CD62L(+);CD3(+);TCRvβ11(+); NKT cells(42.36%), the frequencies of CCR7(+);CD3(+);TCRvα24(+); NKT cells(9.24%) and CCR7(+);CD3(+);TCRvβ11(+); NKT cells(8.22%) were not significant. There were no significant differences in the secretions of IL-4 by CD3(+);TCRvα24(+); NKT cells(13.01%) and CD3(+);TCRvβ11(+); NKT cells(6.62%), and IFN-γ by CD3(+);TCRvα24(+); NKT cells(38.12%) and CD3(+);TCRvβ11(+); NKT cells(26.95%). However, there were significant differences between the mean frequency of IFN-γ(+);IL-4(+);CD3(+);TCRvα24(+); NKT cells(12.65%) and that of IFN-γ(+);IL-4(+);CD3(+);TCRvβ11(+); NKT cells(3.02%).

CONCLUSION

There were some differences between CD3(+);TCRvα24(+); NKT cells and CD3(+);TCRvβ11(+); NKT cells in their frequencies, phenotypes and productions of cytokines. In all, although their frequencies were low, the complicated phenotypes and high secretions of cytokines(IL-4 and IFN-γ) assigned NKT cells immunoregulatory effects.

摘要

目的

阐明CD3(+)；TCRvα24(+)；NKT细胞与CD3(+)；TCRvβ11(+)；NKT细胞在频率、亚群、表型及生物学功能方面的差异，以全面了解NKT细胞在免疫应答中的作用。

方法

分离献血者外周血单个核细胞（PBMCs），采用流式细胞术直接检测或经佛波酯（PMA）加离子霉素刺激后检测细胞表面标志物（CD3、TCRvα24、TCRvβ11、CD4、CD8、CD45RA、CD62L、CCR7）及细胞内细胞因子（IL-4、IFN-γ）。

结果

PBMCs中CD3(+)；TCRvα24(+)；NKT细胞和CD3(+)；TCRvβ11(+)；NKT细胞的平均频率分别为0.63%和0.43%，且存在个体差异。一小部分NKT细胞共表达TCRvα24和TCRvβ11。根据CD4和CD8分子的表达情况，可将人CD3(+)；TCRvα24(+)；NKT细胞亚群分为CD4(+)；NKT 64.35%、CD8(+)；NKT 19.04%、CD4(-)；CD8(-)；NKT 17.18%，CD3(+)；TCRvβ11(+)；NKT细胞亚群分为CD4(+)；NKT 53.69%、CD8(+)；NKT 18.99%、CD4(-)；CD8(-)；NKT 29.74%。相对亚型之间无显著差异。CD45RA(+)；CD3(+)；TCRvβ11(+)；NKT细胞的频率（71.14%）高于CD45RA(+)；CD3(+)；TCRvα24(+)；NKT细胞，二者差异有统计学意义。CD62L(+)；CD3(+)；TCRvα24(+)；NKT细胞的频率（46.26%）与CD62L(+)；CD3(+)；TCRvβ11(+)；NKT细胞的频率（42.36%）之间，CCR7(+)；CD3(+)；TCRvα24(+)；NKT细胞的频率（9.24%）与CCR7(+)；CD3(+)；TCRvβ11(+)；NKT细胞的频率（8.22%）之间差异无统计学意义。CD3(+)；TCRvα24(+)；NKT细胞分泌IL-4的频率（13.01%）与CD3(+)；TCRvβ11(+)；NKT细胞分泌IL-4的频率（6.62%）之间，CD3(+)；TCRvα24(+)；NKT细胞分泌IFN-γ的频率（38.12%）与CD3(+)；TCRvβ11(+)；NKT细胞分泌IFN-γ的频率（26.95%）之间差异无统计学意义。然而，IFN-γ(+)；IL-4(+)；CD3(+)；TCRvα24(+)；NKT细胞的平均频率（12.65%）与IFN-γ(+)；IL-4(+)；CD3(+)；TCRvβ11(+)；NKT细胞的平均频率（3.02%）之间差异有统计学意义。