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Updated: Aug 25, 2026

Generation of Induced Regulatory T Cells from Primary Human Naïve and Memory T Cells
Published on: April 16, 2012
[Regulation of CD3, CD4 and CD8 expressions on PMA-activated human peripheral T cells]
Jian-hua Mao1, Zhi-min Chen, Yong-min Tang
1The Affiliated Children's Hospital, College of Medicine, Zhejiang University, Hangzhou 310003, China.
Objective:
To investigate the effect of mitogen Phorbol 12-myristate 13-Acetate (PMA) on CD3, CD4 and CD8 expression of human T-lymphocytes.
Methods:
Peripheral blood mononuclear cells from 37 blood samples stimulated in vitro with PMA at different concentrations (2,5,10,20 and 50 ng/ml for 4 hours) and time (10 ng/ml for 2,4 and 6 hours) were analyzed by 4-color flow cytometry (FCM).
Results:
Under different PMA stimulation protocols,significant CD4 down-regulation was observed,which was negatively correlated with intracellular cytokine secretion (r= 0.601,P<.001), except for PMA stimulation at 10 ng/ml for 2-hours which showed no significant intracellular cytokine secretion. The expressions of CD3 and CD8 molecules after PMA activation were not significantly affected as compared with pre-activation. Among CD3 positive T lymphocytes, CD4/CD8 double-negative cells only account for 5.52%.
Conclusion:
PMA has a significant down-regulation effect on CD4 molecules of Th cells, without altering the CD3 and CD8 expression. For quantitative analysis of Th1/Th2 variation, indirect method such as CD3(+)CD8( ) T cells can be used to define CD4(+) Th cells stimulated by PMA in the future.
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