Changes of phenotype and function of human CD4 CD25 T cells induced by transfection of Foxp3

Abstract

The aim of this paper is to explore the effects of transfection of Foxp3 gene on the phenotype and function of naive CD4⁺ T cells. The pMSCV-Foxp3 retroviral vector encoding Foxp3 gene was transduced into the PT67 packaging cell line. Virus-containing supernatant was applied to differentiate CD4⁺CD25^- T cells. The resulting cells were sorted with flow cytometry. The expressions of CD25, CD127, CTLA-4 and the proliferation of transfected T cells were examined. The effect of transfected CD4⁺ T cells on the proliferation and cytokine production of CD4⁺CD25^- T cells was examined. Foxp3-gene transfected CD4⁺ T cells could express Foxp3 and transfection of Foxp3 gene up-regulated the expressions of CD25 and CTLA-4, but down-regulated CD127 expression. After transfection, the proliferation of CD4⁺ T cells was eliminated. Transfected T cells inhibited the proliferation of CD4⁺CD25^- T cells. CD4⁺CD25^- T cells acquired a regulatory phenotype and function after it was transduced with the Foxp3 gene. This suggested a key role of Foxp3 in the generation of CD4⁺CD25⁺ regulatory T cells.

Keywords

Forkhead transcription factors / FOXP3 protein, human / T-lymphocytes, regulatory / Retroviridae