CAJ | 학술논문

目的探讨同种异体骨髓间充质干细胞(MSC)体内外对系统性红斑狼疮(SLE)患者外周血CD4+Foxp3+T淋巴细胞及人脐带MSC移植对MRL/lpr鼠脾脏和淋巴结CD4+Foxp3+T淋巴细胞水平的影响.方法血缘相关供者骨髓中分离培养MSC移植治疗5例SLE患者,采用流式细胞术检测移植前后外周血CD4+Foxp3+T淋巴细胞百分率.7例SLE患者外周血单个核细胞(PBMC)分别与SLE患者和正常人骨髓MSC按不同比例体外共培养72 h,检测共培养后PBMC中CD4+Foxp3+T淋巴细胞百分率.MRL/Ipr鼠输注脐带MSC后检测脾脏和淋巴结CD4+Foxp3+T淋巴细胞百分率.结果 SLE患者异基因骨髓MSC移植后1周外周血CD4+Foxp3+T淋巴细胞百分率(4.8±1.6)%和移植后3个月(6.0±2.6)%均较移植前(2.1±1.2)%明显升高(5例,P<0.05).正常骨髓MSC与SLE患者PBMC共培养后CD4+Foxp3+T淋巴细胞百分率明显升高(P<0.05).且存在剂量依赖性,狼疮MSC也可上调SLE患者CD4+Foxp3+T淋巴细胞水平,但作用较正常MSC弱(P<0.05);正常MSC培养上清也可上调SLE患者PBMC中CD4+Foxp3+T淋巴细胞水平,但作用弱于MSC:PBMC=1:1组(P<0.05).MRL/Ipr鼠经1次或3次脐带MSC移植后脾脏CD4+Foxp3+T淋巴细胞百分率均较对照组高(P<0.05),但淋巴结CD+Foxp3+T淋巴细胞百分率均较对照组低(p<0.01),1次和3次移植组间差异无统计学意义.结论异基因甚至异种MSC移植可上调SLE患者或MRL/Ipr鼠CD4+Foxp3+T淋巴细胞水平,同时体外试验也得出相同结论,且体外上调作用呈一定剂量依赖性,CD4+Foxp3+T淋巴细胞水平上调可能是MSC移植治疗SLE有效的机制之一.
목적 탐토동충이체골수간충질간세포(MSC)체내외대계통성홍반랑창(SLE)환자외주혈CD4+Foxp3+T림파세포급인제대MSC이식대MRL/lpr서비장화림파결CD4+Foxp3+T림파세포수평적영향.방법 혈연상관공자골수중분리배양MSC이식치료5례SLE환자,채용류식세포술검측이식전후외주혈CD4+Foxp3+T림파세포백분솔.7례SLE환자외주혈단개핵세포(PBMC)분별여SLE환자화정상인골수MSC안불동비례체외공배양72 h,검측공배양후PBMC중CD4+Foxp3+T림파세포백분솔.MRL/Ipr서수주제대MSC후검측비장화림파결CD4+Foxp3+T림파세포백분솔.결과 SLE환자이기인골수MSC이식후1주외주혈CD4+Foxp3+T림파세포백분솔(4.8±1.6)%화이식후3개월(6.0±2.6)%균교이식전(2.1±1.2)%명현승고(5례,P<0.05).정상골수MSC여SLE환자PBMC공배양후CD4+Foxp3+T림파세포백분솔명현승고(P<0.05).차존재제량의뢰성,랑창MSC야가상조SLE환자CD4+Foxp3+T림파세포수평,단작용교정상MSC약(P<0.05);정상MSC배양상청야가상조SLE환자PBMC중CD4+Foxp3+T림파세포수평,단작용약우MSC:PBMC=1:1조(P<0.05).MRL/Ipr서경1차혹3차제대MSC이식후비장CD4+Foxp3+T림파세포백분솔균교대조조고(P<0.05),단림파결CD+Foxp3+T림파세포백분솔균교대조조저(p<0.01),1차화3차이식조간차이무통계학의의.결론 이기인심지이충MSC이식가상조SLE환자혹MRL/Ipr서CD4+Foxp3+T림파세포수평,동시체외시험야득출상동결론,차체외상조작용정일정제량의뢰성,CD4+Foxp3+T림파세포수평상조가능시MSC이식치료SLE유효적궤제지일.
Objective To investigate the in vivo or in vitro immune regulatory effects of allogeneic bone-marrow mesenchymal stem ceils (MSC) and human umbilical cord MSC on CD4+ Foxp3+ T cells in the peripheral blood of patients with systemic lupus erythematosus (SLE) and in the spleen of MRL/Ipr mice. Methods Human MSC were isolated and expanded from bone marrow cells of healthy donors and infused into five SLE patients. The percentages of CD4+ Foxp3+ T cells in peripheral blood were detected by flow cytometry. Human peripheral blood mononuclear cells (PBMC) were prepared by centrifugation on a Ficoll Hypaque density gradient. The MSC and PBMC from unrelated donors (MSC:PBMC =1:1,1:10,1:50) were added into 24-well plates. After 72h of co-culture, the percentages of CD4+ Foxp3+ T cells were detected by flow cytome- try. Twenty four 18-week-old MRL/Ipr female mice were divided into 3 groups and were injected with umbilical cord MSC (1×106 cells for one time, 1×106 cells for three times and 0.5 ml sodium chloride as control respectively). The percentages of CD4+ Foxp3+ T cells in spleen and lymphoid nodes were detected by flow cytometry. Results The percentages of blood CD4+ Foxp3+ T cells at one week (4.8± 1.6)% and at three months (6.0±2.6)% post MSC transplantation for patients with SLE were both higher than that before transplantation (2.1±1.2)% (n=5,P<0.05). The co-culture of normal bone marrow MSC with PBMC from SLE patients resulted in a statistically significant increase of CD+ Foxp3+ T cells percentage in PBMC on a dose dependent manner (P<0.05). The percentages of CD4+ Foxp3+ T cells of PBMC from SLE patients co-cultured with lupus MSC were lower than that of normal MSC (P<0.05). The cultured supematant of normal MSC also upregulated the percentages of CD4+ Foxp3+ T cells of PBMC from SLE patients (P<0.05). The MRL/lpr mice that had been injected umbilical cord MSC for one time and three times had higher percentages of CD4+ Foxp3+T cells in the spleen but lower in the lymphoid nodes as compared with controls (P<0.01), but without statistical significant difference. Conclusion Allogeneic or heterogeneie MSC transplantation upregulates the percentages of CD4+ Foxp3+ T cells in SLE patients or in MRL/Ipr mice. Upregnlation of Treg population may be one of the mechanisms of MSC transplantation that is effective for SLE treatment.