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Updated: Jul 15, 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
TGF-beta1 modulates Foxp3 expression and regulatory activity in distinct CD4+ T cell subsets
1Department of Microbiology and Immunology and Strategic Training Centre in Infectious Diseases and Autoimmunity, McGill University, Montreal, Quebec, Canada.
Transforming T cells into regulatory cells depends on transforming growth factor-beta 1 (TGF-β1). This cytokine induces forkhead box p3 (Foxp3) expression in CD4(+)CD25(-)CD45RB(LOW) T cells, promoting IL-10-dependent regulatory functions.
Area of Science:
- Immunology
- Cell Biology
- T cell differentiation
Background:
- Forkhead box p3 (Foxp3) is a key transcription factor for CD4(+) regulatory T cells (T(REG)).
- Signals regulating Foxp3 expression and T(REG) cell function remain incompletely understood.
- Transforming growth factor-beta (TGF-β) has been implicated in modulating Foxp3 expression.
Purpose of the Study:
- To investigate the effects of TGF-β1 on the induction and maintenance of regulatory functions in distinct CD4(+) T cell subsets.
- To elucidate the role of TGF-β1 in Foxp3 induction and T(REG) cell differentiation from non-regulatory precursors.
Main Methods:
- Priming of naive CD4(+)CD25(-)CD45RB(HIGH) and CD4(+)CD25(-)CD45RB(LOW) T cells with TGF-β1.
- Assessment of Foxp3 expression (mRNA and protein) and regulatory functions (T cell proliferation suppression) following TCR activation.
- Analysis of IL-10 production and the effect of anti-IL-10 receptor antibodies on T(REG) cell activity.
- Evaluation of TGF-β1 effects on naturally occurring CD4(+)CD25(+) T(REG) cells.
Main Results:
- TGF-β1 induced Foxp3 expression and promoted the differentiation of regulatory T cells from CD4(+)CD25(-) T cells in a concentration-dependent manner.
- Naive CD4(+)CD25(-)CD45RB(HIGH) T cells did not acquire regulatory functions or significant Foxp3 expression upon TGF-β1 treatment.
- CD4(+)CD25(-)CD45RB(LOW) T cells acquired regulatory activity and de novo Foxp3 expression after TGF-β1 exposure, mediating IL-10-dependent suppression.
- TGF-β1 treatment sustained Foxp3 expression in naturally occurring CD4(+)CD25(+) T(REG) cells but did not enhance their expansion or function.
Conclusions:
- TGF-β1 is crucial for inducing IL-10-secreting CD4(+) T(REG) cells from CD4(+)CD25(-)CD45RB(LOW) precursors via de novo Foxp3 production.
- TGF-β1 plays a role in maintaining peripheral homeostasis of natural T(REG) cells by sustaining Foxp3 expression.
- These findings highlight TGF-β1 as a key regulator in adaptive T(REG) cell generation and natural T(REG) cell maintenance.
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