Experimental study on the in vitro induction of regulatory T cells by umbilical cord mesenchymal stem cells with positive human leukocyte antigen-G
10.3969/j.issn.1674-7445.2018.02.002
- VernacularTitle:人类白细胞抗原-G阳性的脐带间充质干细胞体外诱导调节性T细胞的实验研究
- Author:
Jian BAI
1
,
2
;
Li XIAO
;
Lanying MIAO
;
Dayong LIN
;
Hong LIU
;
Yu GAO
;
Wen CHEN
;
Lili BI
;
Xiangrui KONG
;
Haiyan HUANG
;
Bingyi SHI
Author Information
1. 100091 北京,解放军第309医院全军器官移植研究所移植研究室 北京市器官移植与免疫调节重点实验室
2. 辽宁中医药大学
- Keywords:
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
- From:
Organ Transplantation
2018;9(2):97-102
- CountryChina
- Language:Chinese
-
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.
