α-黑素细胞剌激素基因原位转染及B7-H1基因修饰的树突状细胞对小鼠同种异体心脏移植排斥反应的影响
发布时间：2018-10-14 16:03
【摘要】：目的 研究α-黑素细胞刺激素(α-MSH)基因原位转染移植物的抗移植排斥效应以及用B7-H1基因修饰的树突状细胞(DC)作为致耐原诱导移植免疫耐受的作用。方法 采用同种异基因小鼠颈部异位心脏移植模型,将携带α-MSH基因的重组腺相关病毒(rAAV-α-MSH)经主动脉灌注原位转染供者小鼠心脏,再行心脏移植术,观察移植物存活时间及病理改变,测定移植心脏原位的α-MSH表达情况和细胞因子水平。构建携带B7-H1基因的重组腺病毒(rAd-B7-H1),转染未成熟DC,制备B7-H1基因修饰的DC(B7-H1-DC),用流式细胞仪检测B7-H1-DC表型特征,MLR观察对同种异基因T细胞增殖的抑制情况;将BALB/c小鼠来源的B7-H1-DC免疫C57BL/6小鼠后,行心脏移植术,观察移植物存活时间及病理改变,测定血清细胞因子水平。结果 携带α-MSH基因的AAV载体经主动脉灌注原位转染供者小鼠心脏,可延长移植物存活时间,明显减轻移植心脏的病理改变,并促进移植心脏局部的细胞因子格局由Th1型向Th2型偏移,趋化因子水平降低。rAd-B7-H1转染的DC可高表达B7-H1,B7-H1-DC在体外可抑制同种异基因T细胞的增殖,并抑制Th1细胞因子产生;BALB/c小鼠来源的B7-H1-DC经尾静脉注入C57BL/6小鼠体内,可诱导受者对供者的抗原特异性低反应性。移植前7天经尾静脉注射B7-H1-DC可以有意义地延长血管化心脏移植物的存活时间,明显地减轻移植心脏的病理改变。结论α-MSH基因原位转染移植物及用B7-H1基因修饰的DC作为致耐原均具有抗移植排斥效应。
[Abstract]:Objective to study the anti-graft rejection effect of 伪 -melanocyte stimulating hormone (伪-MSH) gene transfection in situ and the effect of B7-H1 gene modified dendritic cells (DC) on the induction of allograft tolerance. Methods the cervical heterotopic heart transplantation model of allogeneic mice was used. The recombinant adeno-associated virus (rAAV- 伪-MSH) carrying 伪-MSH gene was perfused into the aorta of donor mice in situ and then underwent heart transplantation. The survival time and pathological changes of grafts were observed, and the expression of 伪-MSH and cytokine levels in orthotopic heart transplantation were determined. A recombinant adenovirus carrying B7-H1 gene (rAd-B7-H1) was constructed and transfected into immature DC, to prepare DC (B7-H1-DC) modified by B7-H1 gene. The phenotype of B7-H1-DC was detected by flow cytometry, and the proliferation of allogeneic T cells was observed by MLR. B7-H1-DC derived from BALB/c mice was immunized with C57BL/6 mice. The graft survival time and pathological changes were observed and serum cytokines were measured. Results the AAV vector carrying 伪-MSH gene was perfused into the aorta to transfect the donor heart in situ. The graft survival time was prolonged and the pathological changes of the transplanted heart were alleviated. The cytokine pattern of transplanted heart was shifted from Th1 type to Th2 type, and the level of chemokine decreased. DC transfected with rAd-B7-H1 could inhibit the proliferation of allogeneic T cells by overexpression of B7-H1 B7-H1-DC in vitro. B7-H1-DC from BALB/c mice was injected into C57BL/6 mice via tail vein to induce the donor antigen-specific hyporesponsiveness. Injection of B7-H1-DC through caudal vein 7 days before transplantation could significantly prolong the survival time of the vascularized heart grafts and significantly alleviate the pathological changes of the transplanted hearts. Conclusion in situ transfection of 伪-MSH gene and DC modified by B7-H1 gene are resistant to graft rejection.
【学位授予单位】：第二军医大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R392
本文编号：2270949
[Abstract]:Objective to study the anti-graft rejection effect of 伪 -melanocyte stimulating hormone (伪-MSH) gene transfection in situ and the effect of B7-H1 gene modified dendritic cells (DC) on the induction of allograft tolerance. Methods the cervical heterotopic heart transplantation model of allogeneic mice was used. The recombinant adeno-associated virus (rAAV- 伪-MSH) carrying 伪-MSH gene was perfused into the aorta of donor mice in situ and then underwent heart transplantation. The survival time and pathological changes of grafts were observed, and the expression of 伪-MSH and cytokine levels in orthotopic heart transplantation were determined. A recombinant adenovirus carrying B7-H1 gene (rAd-B7-H1) was constructed and transfected into immature DC, to prepare DC (B7-H1-DC) modified by B7-H1 gene. The phenotype of B7-H1-DC was detected by flow cytometry, and the proliferation of allogeneic T cells was observed by MLR. B7-H1-DC derived from BALB/c mice was immunized with C57BL/6 mice. The graft survival time and pathological changes were observed and serum cytokines were measured. Results the AAV vector carrying 伪-MSH gene was perfused into the aorta to transfect the donor heart in situ. The graft survival time was prolonged and the pathological changes of the transplanted heart were alleviated. The cytokine pattern of transplanted heart was shifted from Th1 type to Th2 type, and the level of chemokine decreased. DC transfected with rAd-B7-H1 could inhibit the proliferation of allogeneic T cells by overexpression of B7-H1 B7-H1-DC in vitro. B7-H1-DC from BALB/c mice was injected into C57BL/6 mice via tail vein to induce the donor antigen-specific hyporesponsiveness. Injection of B7-H1-DC through caudal vein 7 days before transplantation could significantly prolong the survival time of the vascularized heart grafts and significantly alleviate the pathological changes of the transplanted hearts. Conclusion in situ transfection of 伪-MSH gene and DC modified by B7-H1 gene are resistant to graft rejection.
【学位授予单位】：第二军医大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R392
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