【摘要】：目的 研究α-黑素細(xì)胞刺激素(α-MSH)基因原位轉(zhuǎn)染移植物的抗移植排斥效應(yīng)以及用B7-H1基因修飾的樹突狀細(xì)胞(DC)作為致耐原誘導(dǎo)移植免疫耐受的作用。方法 采用同種異基因小鼠頸部異位心臟移植模型,將攜帶α-MSH基因的重組腺相關(guān)病毒(rAAV-α-MSH)經(jīng)主動脈灌注原位轉(zhuǎn)染供者小鼠心臟,再行心臟移植術(shù),觀察移植物存活時間及病理改變,測定移植心臟原位的α-MSH表達(dá)情況和細(xì)胞因子水平。構(gòu)建攜帶B7-H1基因的重組腺病毒(rAd-B7-H1),轉(zhuǎn)染未成熟DC,制備B7-H1基因修飾的DC(B7-H1-DC),用流式細(xì)胞儀檢測B7-H1-DC表型特征,MLR觀察對同種異基因T細(xì)胞增殖的抑制情況;將BALB/c小鼠來源的B7-H1-DC免疫C57BL/6小鼠后,行心臟移植術(shù),觀察移植物存活時間及病理改變,測定血清細(xì)胞因子水平。結(jié)果 攜帶α-MSH基因的AAV載體經(jīng)主動脈灌注原位轉(zhuǎn)染供者小鼠心臟,可延長移植物存活時間,明顯減輕移植心臟的病理改變,并促進(jìn)移植心臟局部的細(xì)胞因子格局由Th1型向Th2型偏移,趨化因子水平降低。rAd-B7-H1轉(zhuǎn)染的DC可高表達(dá)B7-H1,B7-H1-DC在體外可抑制同種異基因T細(xì)胞的增殖,并抑制Th1細(xì)胞因子產(chǎn)生;BALB/c小鼠來源的B7-H1-DC經(jīng)尾靜脈注入C57BL/6小鼠體內(nèi),可誘導(dǎo)受者對供者的抗原特異性低反應(yīng)性。移植前7天經(jīng)尾靜脈注射B7-H1-DC可以有意義地延長血管化心臟移植物的存活時間,明顯地減輕移植心臟的病理改變。結(jié)論α-MSH基因原位轉(zhuǎn)染移植物及用B7-H1基因修飾的DC作為致耐原均具有抗移植排斥效應(yīng)。
[Abstract]:Objective to study the anti-graft rejection effect of 偽 -melanocyte stimulating hormone (偽-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- 偽-MSH) carrying 偽-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 偽-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 偽-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 偽-MSH gene and DC modified by B7-H1 gene are resistant to graft rejection.
【學(xué)位授予單位】：第二軍醫(yī)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2006
【分類號】：R392

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本文編號：2270949