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CD4(+)Foxp3(+) regulatory T cells converted by rapamycin from peripheral CD4(+)CD25(-) naive T cells display more potent regulatory ability in vitro
Chen Jian-fei1; Gao Jie1; Zhang Dong2; Wang Zi-han1; Zhu Ji-ye1
关键词Rapamycin Foxp3 b Cells Cd127
刊名CHINESE MEDICAL JOURNAL
2010-04-05
DOI10.3760/cma.j.issn.0366-6999.2010.07.032
123期:7页:942-948
收录类别SCI
文章类型Article
WOS标题词Science & Technology
类目[WOS]Medicine, General & Internal
研究领域[WOS]General & Internal Medicine
关键词[WOS]TRANSCRIPTION FACTOR FOXP3 ; B-CELLS ; GERMINAL-CENTERS ; DENDRITIC CELLS ; CUTTING EDGE ; TOLERANCE ; ANTIGEN ; EXPRESSION ; INDUCTION ; DIFFERENTIATION
英文摘要

Background Rapamycin (RAPA) is a relatively new immunosuppressant drug that functions as a serine/threonine kinase inhibitor to prevent rejection in organ transplantation. RAPA blocks activation of T-effector (Teff) cells by inhibiting the response to interleukin-2. Recently, RAPA was also shown to selectively expand the T-regulator (Treg) cell population. To date, no studies have examined the mechanism by which RAPA converts Teff cells to Treg cells.

Methods Peripheral CD4(+)CD25(-) naive T cells were cultivated with RAPA and B cells as antigen-presenting cells (APCs) in vitro. CD4(+)CD25(-) T cells were harvested after 6 days and analyzed for expression of forkhead box protein 3 (Foxp3) using flow cytometry. CD4(+)CD25(+)CD127(-) subsets as the converted Tregs were isolated from the mixed lymphocyte reactions (MLR) with CD127 negative selection, followed by CD4 and CD25 positive selection using microbeads and magnetic separation column (MSC). Moreover, mRNA was extracted from converted Tregs and C57BL/6 naive CD4(+)CD25(+) T cells and Foxp3 levels were examined by quantitative real-time polymerase chain reaction (rt-PCR). A total of 1x10(5) carboxyfluorescein succinimidyl ester (CFSE)-labeled naive CD4(+)CD25(-) T cells/well from C57BL/6 mice were cocultured with DBA/2 or C3H maturation of dendritic cells (mDCs) (0.25x10(5)/well) in 96-well round-bottom plates for 6 days. Then 1x10(5) or 0.25x10(5) converted Treg cells were added to every well as regulatory cells. Cells were harvested after 6 days of culture and analyzed for proliferation of CFSE-labeled naive CD4(+)CD25(-) T cells using flow cytometry. Data were analyzed using CellQuest software.

Results We found that RAPA can convert peripheral CD4(+)CD25(-) naive T Cells to CD4(+)Foxp3(+) Treg cells using B cells as APCs, and this subtype of Treg can potently suppress left proliferation and maintain antigenic specificity.

Conclusion Our findings provide evidence that RAPA induces Treg cell conversion from Teff cells and uncovers an additional mechanism for tolerance induction by RAPA. Chin Med J 2010;123(7):942-948

语种英语
WOS记录号WOS:000276683700032
项目编号30772050
资助机构National Natural Science Foundation of China
引用统计
被引频次:13[WOS]   [WOS记录]     [WOS相关记录]
文献类型期刊论文
条目标识符http://ir.bjmu.edu.cn/handle/400002259/52846
专题北京大学第二临床医学院_肝胆外科
北京大学基础医学院
作者单位1.Peking Univ, Ctr Transplantat, Dept Hepatobiliary Surg, Peoples Hosp, Beijing 100044, Peoples R China
2.UPMC, Div Plast & Reconstruct Surg, Dept Surg, Starzl Transplantat Inst, Pittsburgh, PA 15261 USA
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Chen Jian-fei,Gao Jie,Zhang Dong,et al. CD4(+)Foxp3(+) regulatory T cells converted by rapamycin from peripheral CD4(+)CD25(-) naive T cells display more potent regulatory ability in vitro[J]. CHINESE MEDICAL JOURNAL,2010,123(7):942-948.
APA Chen Jian-fei,Gao Jie,Zhang Dong,Wang Zi-han,&Zhu Ji-ye.(2010).CD4(+)Foxp3(+) regulatory T cells converted by rapamycin from peripheral CD4(+)CD25(-) naive T cells display more potent regulatory ability in vitro.CHINESE MEDICAL JOURNAL,123(7),942-948.
MLA Chen Jian-fei,et al."CD4(+)Foxp3(+) regulatory T cells converted by rapamycin from peripheral CD4(+)CD25(-) naive T cells display more potent regulatory ability in vitro".CHINESE MEDICAL JOURNAL 123.7(2010):942-948.
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