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[Differences between CD3⁺ TCRvα24⁺ NKT cell and CD3⁺ TCRvβ11⁺ NKT cell in PBMC]
Zi-Tao Li1, Bin-Yan Yang, Chang-You Wu
1Institute of Immunology, Zhongshan School of Medicine, State Ministry of Education Key Laboratory of Tropical Disease Control Research, Sun Yat-sen University, Guangzhou 510080, China.
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.
