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Updated: Jun 9, 2026
In Vitro Differentiation of Human CD4+FOXP3+ Induced Regulatory T Cells (iTregs) from Naïve CD4+ T Cells Using a TGF-β-containing Protocol
Published on: December 30, 2016
[Changes of Foxp3 and IL-10 and TGF-beta in aging of mice]
Yi Zhao1, Yan Wang, Ji-Zhen Liu
1Department of Microbiology and Immunology, Guangdong Medical College, Dongguan 523808, China. zhaoyicomnet@yahoo.com.cn
Aim:
To compare the expression levels of Foxp3 mRNA in the spleen PBMC (peripheral blood mononuclear cell) in the progress of aging mice and analyse the relevance with serum IL-10 and TGF-beta.
Methods:
Healthy male BALB/c mice were divided into 3 groups at random (8 mice each group): 2m group with 2 months mice (young), 6m group with 6 months mice (middle-aged) and 15m group with 15 months mice (aged). The phenotype of CD4(+);CD25(+);Foxp3(+); T cells derived from normal murine SPL were analyzed by FCM. Real-time fluorescence quantitative RT-PCR was used to determine the expression levels of Foxp3 mRNA in the spleen PBMC. ELISA was used to detect the concentration of IL-10 and TGF-beta.
Results:
The subsets of CD4(+);CD25(+);Foxp3(+); T lymphocytes from murine splenocytes were significantly higher in 15m group than that in 2m group(P<0.01). The expression levels of Foxp3 mRNA in the spleen PBMC were significantly higher in 15m group than that in 2m group(P<0.01).The serum concentration of IL-10 and TGF-beta were significantly higher in 15m group than that in 2m group(P<0.01).There was no correlation between serum concentration of IL-10 and TGF-beta and Foxp3 mRNA expression levels in the spleen PBMC in 15m group.
Conclusion:
The level of the CD4(+);CD25(+);Foxp3(+); T cells and Foxp3 mRNA expression levels were up-regulated and IL-10 and TGF-beta contents were increased in the progress of aging mice. Foxp3 and IL-10 and TGF-beta may be play a part in the process of immunosenescence. Quantitative detection of Foxp3 mRNA by the method of SYBR Green I real-time fluorescence quantitative RT-PCR is stable and reliable.