Phenotypic diversity of human nature and induced CD4+CD25+FoxP3+CD127- regulatory T cells

doi:10.3969/j.issn.2095-4344.2016.02.015

Chinese Journal of Tissue Engineering Research ›› 2016, Vol. 20 ›› Issue (2): 236-241.doi: 10.3969/j.issn.2095-4344.2016.02.015

Previous Articles Next Articles

Phenotypic diversity of human nature and induced CD4+CD25+FoxP3+CD127- regulatory T cells

Wang Hai-hao¹, Zhu Li², Yang Pei-wen³, Guo Qian-nan¹

¹Cardiopulmonary Transplantation Institute of Tongji Hospital, Department of Cardiovascular Surgery, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, Hubei Province, China;²Department of Hematology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, Hubei Province, China; ³Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, Hubei Province, China

Received:2015-11-15 Online:2016-01-08 Published:2016-01-08
About author:Wang Hai-hao, M.D., Attending physician, Cardiopulmonary Transplantation Institute of Tongji Hospital, Department of Cardiovascular Surgery, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, Hubei Province, China
Supported by:
the Scientific Research Foundation for the Returned Overseas Chinese Scholars, State Education Ministry of China, No. (2014)1685; the Independent Innovation Research Fund of Huazhong University of Science and Technology, No. 2013QN203

Abstract

Abstract:

BACKGROUND: Regulatory T cells (Treg) are classified into two subsets, nature Treg (nTreg) and induced Treg (iTreg). Although there is consensus that CD4+CD25+FoxP3+CD127- is the widely accepted phenotype of Treg, it remains unclear what is the difference in phenotypes including cytokine patterns of nTreg and iTreg.
OBJECTIVE: To understand and compare the plasticity of nTreg and iTreg and to search the exact mechanism of cytokine secretion in Tregs.
METHODS: We investigated the frequency and cytokine pattern of CD4+CD25+FoxP3+CD127-nTreg in freshly separated peripheral blood mononuclear cells of five healthy individuals using 8-color fluorescence flow cytometry (FACSCanto II). Subsequently, after 9 days of allostimulation in mixed lymphocytes, the frequency and cytokine pattern of CD4+CD25+FoxP3+CD127-iTreg were determined and analyzed.
RESULTS AND CONCLUSION: In fresh cells, (1.5±0.70)% of CD4+ T cells were CD4+CD25+FoxP3+CD127- nTregs. Almost all these cells expressed interferon (IFN)-γ-, interleukin (IL)-2- or transforming growth factor-β+, and partial cells expressed IL-10+ or IL-10-. After 9-day allostimulation, the number of CD4+CD25+FoxP3+CD127- iTreg expressing IFN-γ+, IL-2-, IL-2+, IL-10+ or TGF-β+ increased strongly. The main subsets of human nTregs were CD4+CD25+FoxP3+CD127-IFN-γ-IL-2-IL-10+TGF-β+ and CD4+CD25+FoxP3+CD127-IFN-γ-IL-2-IL-10- TGF-β+ T cells. The proportion of each subset in CD4+ T cells was (1.1±0.59)% and (0.39±0.16)%, respectively. Whereas the main subsets of human iTregs were CD4+CD25+FoxP3+CD127-IFN-γ+IL-2-IL-10+TGF-β+ and CD4+CD25+FoxP3+CD127-IFN-γ+IL-2+IL-10+TGF-β+. Human nTregs were characterized as IFN-γ-IL-2- double negative, producing IL-10 and TGF-β or only TGF-β without IL-10, and not proliferating in vitro. During the allostimulation in mixed lymphocytes, IFN-γ+ iTregs proliferated remarkably. One-third of IFN-γ+ iTreg expressed IL-2+, and two-thirds of IFN-γ+ iTregs expressed IL-2, both of which produce IL-2 and TGF-β. Our results imply that CD4+CD25+FoxP3+CD127- Treg are potentially immunosuppressive probably because of their mandatory TGF-β and optional IL-10 production.

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

Wang Hai-hao, Zhu Li, Yang Pei-wen, Guo Qian-nan. Phenotypic diversity of human nature and induced CD4+CD25+FoxP3+CD127- regulatory T cells[J]. Chinese Journal of Tissue Engineering Research, 2016, 20(2): 236-241.

Figures/Tables 4

References

[1] Sakaguchi S, Miyara M, Costantino CM.et al.FOXP3⁺ regulatory T cells in the human immune system. Nat Rev Immunol.2010;10(7):490-500.

[2] Sakaguchi S, Powrie F.Emerging challenges in regulatory T cell function and biology. Science.2007;317(5838):627-629.

[3] Sakaguchi S, Wing K, Yamaguchi T.Dynamics of peripheral tolerance and immune regulation mediated by Treg.Eur J Immunol.2009；39(9):2331-2336.

[4] Jones E, Dahm-Vicker M, Golgher D,et al.CD25⁺ regulatory T cells and tumor immunity. Immunol Lett.2003;85(2):141-143.

[5] Adeegbe D, Bayer AL, Levy RB,et al.Cutting edge: allogeneic CD4⁺CD25⁺Foxp3⁺ T regulatory cells suppress autoimmunity while establishing transplantation tolerance. J Immunol.2006;176(12):7149-7153.

[6] Bluestone JA, Abbas AK. Natural versus adaptive regulatory T cells. Nat Rev Immunol.2003;3(3):253-257.

[7] Cassis L, Aiello S, Noris M.Natural versus adaptive regulatory T cells. Contrib Nephrol.2005;146:121-131.

[8] Sakaguchi S. The origin of FOXP3-expressing CD4⁺ regulatory T cells: thymus or periphery. J Clin Invest. 2003; 112(9):1310-1312.

[9] Sakaguchi S, Sakaguchi N, Asano M,et al.Immunologic self-tolerance maintained by activated T cells expressing IL-2 receptor alpha-chains (CD25). Breakdown of a single mechanism of self-tolerance causes various autoimmune diseases. J Immunol.1995;155(3):1151-1164.

[10] Baecher-Allan C, Brown JA, Freeman GJ,et al.CD4⁺CD25 high regulatory cells in human peripheral blood. J Immunol. 2001;167(3):1245-1253.

[11] Hori S, Nomura T, Sakaguchi S.Control of regulatory T cell development by the transcription factor Foxp3. Science. 2003;299(5609):1057-1061.

[12] Fontenot JD, Gavin MA, Rudensky AY.Foxp3 programs the development and function of CD4⁺CD25⁺ regulatory T cells. Nat Immunol.2003;4(4):330-336.

[13] Khattri R, Cox T, Yasayko SA,et al.An essential role for Scurfin in CD4⁺CD25⁺ T regulatory cells. Nat Immunol. 2003; 4(4):337-342.

[14] Gambineri E, Torgerson TR, Ochs HD.Immune dysregulation, polyendocrinopathy, enteropathy, and X-linked inheritance (IPEX), a syndrome of systemic autoimmunity caused by mutations of FOXP3, a critical regulator of T-cell homeostasis. Curr Opin Rheumatol.2003; 15(4):430-435.

[15] Bennett CL, Christie J, Ramsdell F,et al.The immune dysregulation, polyendocrinopathy, enteropathy, X-linked syndrome (IPEX) is caused by mutations of FOXP3. Nat Genet.2001;27(1):20-21.

[16] Liu W1, Putnam AL, Xu-Yu Z,et al.CD127 expression inversely correlates with FoxP3 and suppressive function of human CD4⁺ T reg cells.J Exp Med.2006;203(7):1701-1711.

[17] Sanchez-Lockhart M, Marin E, Graf B,et al.Cutting edge: CD28-mediated transcriptional and posttranscriptional regulation of IL-2 expression are controlled through different signaling pathways. J Immunol.2004; 173(12):7120-7124.

[18] Thornton AM, Donovan EE, Piccirillo CA,et al.Cutting edge: IL-2 is critically required for the in vitro activation of CD4⁺CD25⁺ T cell suppressor function. J Immunol.2004;172(11):6519-6523.

[19] Wu H, Li P, Shao N,et al.Aberrant expression of Treg-associated cytokine IL-35 along with IL-10 and TGF-beta in acute myeloid leukemia. Oncol Lett.2012;3(5):1119-1123.

[20] Daniel V, Naujokat C, Sadeghi M,et al.Observational support for an immunoregulatory role of CD3⁺CD4⁺CD25⁺IFN^-gamma⁺ blood lymphocytes in kidney transplant recipients with good long-term graft outcome. Transpl Int.2008;21(7):646-660.

[21] Daniel V, Sadeghi M, Wang H,et al.CD4⁺CD25⁺Foxp3⁺IFN-gamma⁺ human induced T regulatory cells are induced by interferon-gamma and suppress alloresponses nonspecifically. Hum Immunol.2011; 72(9):699-707.

[22] 王海灏,仰霈雯,雷家慧.两种多克隆刺激条件下人诱导性Treg表型的多样性变化[J]. 中国免疫学杂志,2012, 28(9):769-774.

[23] Roussel M, Davis BH, Fest T,et al.Toward a reference method for leukocyte differential counts in blood: Comparison of three flow cytometric candidate methods. Cytometry A. 2012;81(11): 973-982.

[24] Gill N, Chenoweth MJ, Verdu EF,et al.NK cells require type I IFN receptor for antiviral responses during genital HSV-2 infection. Cell Immunol.2011;269(1):29-37.

[25] Daniel V, Sadeghi M, Wang H,et al.CD4(⁺)CD25(⁺)Foxp3(⁺) IFNgamma(⁺) Treg are immunosuppressive in vitro and increase with intensity of the alloresponse in pretransplant MLC. Transpl Immunol. 2012;27(2-3):114-121.

[26] Daniel V, sadeghi M, Wang H,et al.In-vitro inhibition of IFNgamma⁺ iTreg mediated by monoclonal antibodies against cell surface determinants essential for iTreg function. BMC Immunol.2012;13(1):47.

Phenotypic diversity of human nature and induced CD4+CD25+FoxP3+CD127- regulatory T cells

PDF

Knowledge

Abstract

Cite this article

share this article

Figures/Tables 4

References

Related Articles 0

Recommended Articles

Metrics

Comments