Experimental study on the <i>in vitro</i> induction of regulatory T cells by umbilical cord mesenchymal stem cells with positive human leukocyte antigen-G

Bai Jian; Xiao Li; Miao Lanying; Lin Dayong; Liu Hong; Gao Yu; Chen Wen; Bi Lili; Kong Xiangrui; Huang Haiyan; Shi Bingyi

doi:10.3969/j.issn.1674-7445.2018.02.002

Volume 9 Issue 2

Mar. 2018

Turn off MathJax

Article Contents

Article Navigation > ORGAN TRANSPLANTATION > 2018 > 9(2): 97-102

Bai Jian, Xiao Li, Miao Lanying, et al. Experimental study on the in vitro induction of regulatory T cells by umbilical cord mesenchymal stem cells with positive human leukocyte antigen-G[J]. ORGAN TRANSPLANTATION, 2018, 9(2): 97-102. doi: 10.3969/j.issn.1674-7445.2018.02.002

Citation:

Bai Jian, Xiao Li, Miao Lanying, et al. Experimental study on the in vitro induction of regulatory T cells by umbilical cord mesenchymal stem cells with positive human leukocyte antigen-G[J]. ORGAN TRANSPLANTATION, 2018, 9(2): 97-102. doi: 10.3969/j.issn.1674-7445.2018.02.002

Citation:

PDF( 1365 KB)

Experimental study on the in vitro induction of regulatory T cells by umbilical cord mesenchymal stem cells with positive human leukocyte antigen-G

doi: 10.3969/j.issn.1674-7445.2018.02.002

Basic Research Laboratory of Organ Transplant Institute, the 309^th Hospital of Chinese People's Liberation Army, Beijing Key Laboratory of Organ Transplantation and Immunology Regulatory, Beijing 100091, China

More Information

Corresponding author: Shi Bingyi, Email: shibingyi@medmail.com.cn
Received Date: 2017-11-01
Available Online: 2021-01-19
Publish Date: 2018-03-15

Abstract

Abstract

Objective To explore the effect of umbilical cord mesenchymal stem cells with positive human leukocyte antigen (HLA)-G on inducing the production of regulatory T cells (Treg) in vitro. Methods Umbilical cord mesenchymal stem cells were isolated from umbilical cord of neonates. PEGFP-N1-HLA-G plasmid was transfected into the human umbilical cord mesenchymal stem cells by liposome transfection, as PEGFP-N1-HLA-G group. PEGFP-N1 empty vector plasmid was transfected into the human umbilical cord mesenchymal stem cells, as PEGFP-N1 group. The human umbilical cord mesenchymal stem cells without empty vector under the same conditions were set as blank control group. Markers of the umbilical cord mesenchymal stem cells were detected using flow cytometry. The expression of HLA-G protein in each group of cells was identified by Western Blot. After mixed-culturing with CD4⁺T cells in peripheral blood of healthy subjects for 24 h and 48 h, the proportion of CD4⁺ CD25⁺ Foxp3⁺Treg in total T cells of each group was detected by flow cytometry. Results CD45, CD34 and HLA-DR presented negative expression on umbilical cord mesenchymal stem cells, while CD29, CD44 and CD105 presented positive expression. HLA-G protein could be expressed in the PEGFP-N1-HLA-G group, which had statistically significant difference compared with the blank control group and PEGFP-N1 group (both P < 0.01). After PEGFP-N1-HLA-G group and CD4⁺T cells were mixed-cultured for 24 h and 48 h, CD4⁺ CD25⁺ Foxp3⁺Treg accounted for (15.3±1.9)% and (14.3±2.1)% of the total T cells respectively, both of which presented statistically significant difference compared with the blank control group and PEGFP-N1 group (all P < 0.05). Conclusions Umbilical cord mesenchymal stem cells with HLA-G gene modified can effectively induce the production of CD4⁺ CD25⁺ Foxp3⁺Treg in vitro.
- Human leukocyte antigen (HLA)-G,
- Umbilical cord mesenchymal stem cells,
- T cells,
- Regulatory T cell (Treg),
- Transfection,
- Modification,
- Immunologic rejection,
- Immunologic tolerance,
- CD4⁺T cells

FullText(HTML)

References(17)

References

[1]	孔伟浩, 张剑.PD-1/PD-L1信号通路在不同组织来源间充质干细胞中免疫调节作用的研究进展[J].器官移植, 2017, 8(6): 483-485.DOI: 10.3969/j.issn.1674-7445.2017.06.015. KONG WH, ZHANG J. Research progress of PD-1/PD-L1 signaling pathway in immunoregulation of mesenchymal stem cell derived from different tissues[J]. Organ Transplant, 2017, 8(6): 483-485.DOI: 10.3969/j.issn.1674-7445.2017.06.015.
[2]	申升, 刘东, 黎程, 等.外周血CD4⁺ CD25^high调节性T细胞比例变化在肾移植受者移植后免疫监测中的应用[J].中国组织工程研究, 2009, 13(18): 3425-3428. DOI: 10.3969/j.issn.1673-8225.2009.18.005. SHEN S, LIU D, LI C, et al. The application of the change of CD4⁺ CD25^high regulatory T cell ratio in peripheral blood of renal transplant recipients in immunological monitoring after transplantation[J]. Chin J Tissue Eng Res, 2009, 13(18): 3425-3428. DOI: 10.3969/j.issn.1673-8225.2009.18.005.
[3]	SHAN J, GUO Y, LUO L, et al. Do CD4⁺Foxp3⁺Treg cells correlate with transplant outcomes: a systematic review on recipients of solid organ transplantation[J]. Cell Immunol, 2011, 270(1): 5-12. DOI: 10.1016/j.cellimm.2011.05.006.
[4]	尹东亮, 孙翀, 朱焕斌, 等.CTLA4-Ig基因修饰骨髓间充质干细胞抑制大鼠肝移植排斥反应[J].器官移植, 2014, 5(4): 231-236. DOI: 10.3969/j.issn.1674-7445. 2014.04.008. YIN DL, SUN C, ZHU HB, et al. Bone marrow mesenchymal stem cell modified by CTLA4-Ig gene can inhibit the rejection of liver transplantation in rats[J]. Organ Transplant, 2014, 5(4): 231-236.DOI: 10.3969/j.issn.1674-7445.2014.04.008.
[5]	WADA N, BARTOLD PM, GRONTHOS S. Human foreskin fibroblasts exert immunomodulatory properties by a different mechanism to bone marrow stromal/stem cells[J]. Stem Cells Dev, 2011, 20(4): 647-659. DOI: 10.1089/scd.2010.0246.
[6]	牛坚, 刘斌, 张业伟, 等.hIL-10修饰骨髓间充质干细胞对大鼠原位异种肝移植急性排斥反应的影响[J].中华肝胆外科杂志, 2010, 16(10): 777-780. DOI: 10.3760/cma.j.issn.1007-8118.2010.10.020. NIU J, LIU B, ZHANG YW, et al. The role of hIL-10-MSCs in immune tolerance of rat discordant liver xenotransplantation[J].Chin J Hepatobil Surg, 2010, 16(10): 777-780. DOI: 10.3760/cma.j.issn.1007-8118.2010.10.020.
[7]	XU DP, SHI WW, ZHANG TT, et al. Elevation of HLA-G-expressing DC-10 cells in patients with gastric cancer[J]. Hum Immunol, 2016, 77(9): 800-804. DOI: 10.1016/j.humimm.2016.01.003.
[8]	马锡慧, 肖漓, 冯凯, 等. 人脐带和骨髓间充质干细胞体外分离培养及生物学特性比较[J/CD]. 中华细胞与干细胞杂志(电子版), 2015, 5(2): 10-13. DOI: 10.3877/cma.j.issn.2095-1221.2015.02.002. MA XH, XIAO L, FENG K, et al. Comparison of biological characteristics of human umbilical cord mesenchymal stem cells with human bone marrow mesenchymal stem cells[J/CD]. Chin J Cell Stem Cell(Electr Edit), 2015, 5(2): 10-13. DOI: 10.3877/cma.j.issn.2095-1221.2015.02.002.
[9]	PLAIN KM, VERMA ND, TRAN GT, et al. Cytokines affecting CD4(+) T regulatory cells in transplant tolerance. interleukin-4 does not maintain alloantigen specific CD4(+) CD25(+) Treg[J]. Transpl Immunol, 2013, 29(1/2/3/4): 51-59. DOI: 10.1016/j.trim.2013.10.003.
[10]	ZHANG C, SHAN J, HUANG Y, et al. Rapamycin in combination with donor-specific CD4⁺ CDCD25⁺Treg cells amplified in vitro might be realize the immune tolerance in clinical organ transplantation[J].Cell Immunol, 2010, 264(2): 111-113. DOI: 10.1016/j.cellimm.2010.05.014.
[11]	WANG Z, XIAO L, SHI BY, et al. Short-term anti-CD25 monoclonal antibody treatment and neogenetic CD4(+)CD25(high) regulatory T cells in kidney transplantation.[J]. Transpl Immunol, 2008, 19(1): 69-73. DOI: 10.1016/j.trim.2008.01.005.
[12]	WANG Z, FU XY, SHI BY, et al. Antigen-specific suppression by induced CD4⁺CD25^high regulatory T cells in kidney recipients[J]. Transplant Proc, 2009, 41(5): 1574-1576. DOI: 10.1016/j.transproceed.2009.01.093.
[13]	徐小丽, 邹萍, 游泳, 等.异基因造血干细胞移植受者外周血中可溶性HLA-G与急性GVHD的相关性[J].中华器官移植杂志, 2010, 31(4): 234-237. DOI: 10.3760/cma.j.issn.0254-1785.2010.04.011. XU XL, ZOU P, YOU Y, et al. The correlation between soluble HLA-G and acute graft-versus-host disease after allo-geneic hematopoietic stem cell transplantation[J].Chin J Organ Transplant, 2010, 31(4): 234-237.DOI: 10.3760/cma.j.issn.0254-1785.2010.04.011.
[14]	WLASIUK P, STEC A, PIECHNIK A, et al. Expression of soluble HLA-G in multiple myeloma patients and patients with renal failure[J]. Leuk Res, 2012, 36(7): 881-883. DOI: 10.1016/j.leukres.2012.02.015.
[15]	余碧珍, 杜庆华, 李庆山, 等.allo-HSCT后外周血TH17细胞/Treg细胞的平衡与aGVHD的相关性[J].中华器官移植杂志, 2015, 36(3): 171-175. DOI: 10.3760/cma.j.issn.0254-1785.2015.03.009. YU BZ, DU QH, LI QS, et al. Relationship between helper T cell 17/regulatory T cells balance and acute graft versus host disease after allogeneic hematopoietic stem cell transplantation[J].Chin J Organ Transplant, 2015, 36(3): 171-175. DOI: 10.3760/cma.j.issn.0254-1785.2015.03.009.
[16]	LEE MW, CHOI J, YANG MS, et al. Mesenchymal stem cells from cryopreserved human umbilical cord blood[J]. Biochem Biophys Res Commun, 2004, 320(1): 273-278. DOI: 10.1016/j.bbrc.2004.04.206.
[17]	BOISSEL L, TUNCER HH, BETANCUR M, et al. Umbilical cord mesenchymal stem cells increase expansion of cord blood natural killer cells[J]. Biol Blood Marrow Transplant, 2008, 14(9): 1031-1038. DOI: 10.1016/j.bbmt.2008.06.016.

Relative Articles

Supplements(0)

Cited By

Proportional views

Proportional views

通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

Figures(2)

Get Citation

PDF

XML

Article Metrics

Article views (200) PDF downloads(10)

Experimental study on the in vitro induction of regulatory T cells by umbilical cord mesenchymal stem cells with positive human leukocyte antigen-G

doi: 10.3969/j.issn.1674-7445.2018.02.002

Abstract

References

Proportional views

Catalog

通讯作者: 陈斌, bchen63@163.com

Article Metrics

Proportional views

Related

Experimental study on the in vitro induction of regulatory T cells by umbilical cord mesenchymal stem cells with positive human leukocyte antigen-G

doi: 10.3969/j.issn.1674-7445.2018.02.002

Abstract

References

Proportional views

Catalog

通讯作者: 陈斌, bchen63@163.com

Article Metrics

Proportional views

Related

Export File

Citation

Format

Content