PTD-hFOXP3 protein acts as an immune regulator to convert human CD4(+)CD25(-) T cells to regulatory T-like cells
Copyright © 2012 Wiley Periodicals, Inc..
Regulatory T cells (Tregs) are critical for maintaining self-tolerance and homeostasis, and have potential application in clinical disease therapy, such as autoimmune diseases and transplant rejection, but their numbers are limited. FOXP3 is a key transcription factor controlling Tregs development and function. Although transfection of CD4(+)CD25(-) lymphocytes with the FOXP3 gene can convert them to Treg-like cells, there is the risk of insertional mutagenesis and thus an alternative to genetic intervention is sought. The protein transduction domain (PTD) from the HIV transactivator of transcription is a useful tool to deliver protein to the cytoplasm and nucleus. In this study, we generated a fusion protein linking the human FOXP3 to PTD (PTD-hFOXP3), and explored its function in T cells. The results showed that the PTD rapidly and effectively delivered the hFOXP3 protein into cells where it localized not only in the cytoplasm, but also to the nucleus. PTD-hFOXP3-transduced Jurkat cells (human T lymphoma cell line) and CD4(+)CD25(-) T cells failed to proliferate and produce IL-2 and IFN-γ, but produced large amounts of the cytokines IL-4, IL-10, and TGF-β, in response to TCR stimulation in vitro. PTD-hFOXP3-transduced CD4(+)CD25(-) T cells also expressed high levels of CTLA-4 and low levels of CD25 after stimulation. Most importantly, PTD-hFOXP3-transduced T cells inhibited the proliferation of activated CD4(+)CD25(-) T cells. Furthermore, chromatin immunoprecipitation assays demonstrated that PTD-hFOXP3 can bind with the IL-2 gene promoter and repress the expression of IL-2. These results indicate that PTD-hFOXP3 has the capability to convert conventional T cells to Treg-like cells.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2012 |
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| Erschienen: |
2012 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:113 |
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| Enthalten in: |
Journal of cellular biochemistry - 113(2012), 12 vom: 01. Dez., Seite 3797-809 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Liu, Xia [VerfasserIn] |
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| Links: |
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| Anmerkungen: |
Date Completed 08.03.2013 Date Revised 12.10.2012 published: Print Citation Status MEDLINE |
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| doi: |
10.1002/jcb.24255 |
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| Förderinstitution / Projekttitel: |
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| 245 | 1 | 0 | |a PTD-hFOXP3 protein acts as an immune regulator to convert human CD4(+)CD25(-) T cells to regulatory T-like cells |
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| 500 | |a Date Revised 12.10.2012 | ||
| 500 | |a published: Print | ||
| 500 | |a Citation Status MEDLINE | ||
| 520 | |a Copyright © 2012 Wiley Periodicals, Inc. | ||
| 520 | |a Regulatory T cells (Tregs) are critical for maintaining self-tolerance and homeostasis, and have potential application in clinical disease therapy, such as autoimmune diseases and transplant rejection, but their numbers are limited. FOXP3 is a key transcription factor controlling Tregs development and function. Although transfection of CD4(+)CD25(-) lymphocytes with the FOXP3 gene can convert them to Treg-like cells, there is the risk of insertional mutagenesis and thus an alternative to genetic intervention is sought. The protein transduction domain (PTD) from the HIV transactivator of transcription is a useful tool to deliver protein to the cytoplasm and nucleus. In this study, we generated a fusion protein linking the human FOXP3 to PTD (PTD-hFOXP3), and explored its function in T cells. The results showed that the PTD rapidly and effectively delivered the hFOXP3 protein into cells where it localized not only in the cytoplasm, but also to the nucleus. PTD-hFOXP3-transduced Jurkat cells (human T lymphoma cell line) and CD4(+)CD25(-) T cells failed to proliferate and produce IL-2 and IFN-γ, but produced large amounts of the cytokines IL-4, IL-10, and TGF-β, in response to TCR stimulation in vitro. PTD-hFOXP3-transduced CD4(+)CD25(-) T cells also expressed high levels of CTLA-4 and low levels of CD25 after stimulation. Most importantly, PTD-hFOXP3-transduced T cells inhibited the proliferation of activated CD4(+)CD25(-) T cells. Furthermore, chromatin immunoprecipitation assays demonstrated that PTD-hFOXP3 can bind with the IL-2 gene promoter and repress the expression of IL-2. These results indicate that PTD-hFOXP3 has the capability to convert conventional T cells to Treg-like cells | ||
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| 650 | 7 | |a tat Gene Products, Human Immunodeficiency Virus |2 NLM | |
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| 700 | 1 | |a Lin, Xin |e verfasserin |4 aut | |
| 700 | 1 | |a Tian, Yuxiang |e verfasserin |4 aut | |
| 700 | 1 | |a Ji, Baoju |e verfasserin |4 aut | |
| 700 | 1 | |a Xia, Sheng |e verfasserin |4 aut | |
| 700 | 1 | |a Xu, Sanrong |e verfasserin |4 aut | |
| 700 | 1 | |a Yin, Qing |e verfasserin |4 aut | |
| 700 | 1 | |a Zhang, Miaomiao |e verfasserin |4 aut | |
| 700 | 1 | |a Jiao, Zhijun |e verfasserin |4 aut | |
| 700 | 1 | |a Wang, Shengjun |e verfasserin |4 aut | |
| 700 | 1 | |a Xu, Huaxi |e verfasserin |4 aut | |
| 700 | 1 | |a Shao, Qixiang |e verfasserin |4 aut | |
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