【摘要】 目的了解病毒白细胞介素10(vIL10)基因对同种小鼠移植心脏的影响。方法用小鼠干细胞病毒(MSCVneo)逆转录病毒表达系统和vIL10cDNA构建MSCVneovIL10重组体,在体外转导CBA(H2K)小鼠的造血干细胞,输入经致死量照射(900rad)的同基因CBA(H2K)小鼠体内,8周后以vIL10表达阳性的CBA(H2K)小鼠(12只,实验组)作为受体,C57BL/6(H2b)小鼠为供体,进行同种腹部异位心脏移植,以未处理、移植无病毒转染的造血干细胞(HSCs)、移植空MSCVneo转染的HSCs鼠(均为10只)为对照组。在移植后第8天,每组受体动物各杀死5只,取出移植心脏行组织病理学、CD+4与CD+8T淋巴细胞浸润(免疫荧光抗体法)检查,IL2、IL4、IL6、小鼠IL10(mIL10)、γ干扰素(IFNγ)、诱导型一氧化氮合酶(iNOS)、B71、B72mRNA表达的测定[逆转录聚合酶链反应(RTPCR)法],余下的动物观察移植心脏存活时间。结果(1)实验组移植心脏存活时间(800±333)d,其他对照组分别为(104±10),(116±11)与(112±17)d(P<001);(2)实验组心肌炎性细胞浸润稀少、心肌结构完整、无明显血管炎,急性排斥反应Ⅰ级,而其他对照组则出现严重的炎性细胞浸润、心肌灶性坏死、心内膜炎、血管炎和心肌出血,急性排斥反应均为Ⅲ级;(3)实验组移植物内IL2、IFNγ、B71、和B72和iNOS的mRNA表达明显下调,而其更多还原

【Abstract】 Objective To prolong murine heart allograft by modificating hematopoietic stem cells with virus interleukin-10（vIL-10）. Methods The recombinant of murine stem cell virus（ MSCVneo） ^vIL-10 was composed of MSCVneo and vIL-10 cDNA and transduced hematopoietic stem cells from CBA（H-2K）mice’s bone marrow in vitro. The transduced hematopoietic stem cells were transplanted into a syngenic CBA（H-2K）mouse with lethal irradiation （900 rad） in the same day through penis vein. The mouse’s heterotopic heart transplantation was conducted using CBA（H-2K） mice as recipients, which vIL-10 in serum were positive by enzyme-linked immunosorbent assay, and donors hearts from C57BL/6（H-2b） mice. Five animals in each group were sacrificed to test histopathology changes, the expression of interleukin ^IL-2, IL-4,IL-6,mIL-10,interferon（IFN）-γ, inducible nitric oxide synthase （iNOS）,B7-1,B7-2 and CD+₄ and CD+₈ T cells subset infiltration in heart transplants with reverse transcriptase polymerase chain reaction , immunohistochemistry and regular pathology. Results Survival time of mice’s allografts experimental group was （80.0±33.3）days. And survival time of control groups were（10.4±1.0）days, （11.6±1.1）days and（ 11.2±1.7）days, respectively ^P<0.01. Heart transplants from experimental group were characterized by sparse lymphocytes infiltration, mild endocarditis and vasculitis and preserved myocardial architecture, which had acute rejection of grade I. Cardiac allografts from other control groups developed severe cellular rejection with severe infiltrating lymphocytes, myocyte injury and necrosis, interstitial edema and hemorrhage, which had acute rejection of grade III.. The expression of IL-2, INF-γ, B7-1, B7-2 and iNOS mRNA in allografts in experimental group markedly down-regulated, whereas that in allografts in control groups markedly upregulated （P<0.05）. CD+₄ and CD+₈ T cell subsets infiltration in heart transplants from experimental group decreased, and that in control groups increased （P<0.05）.Conclusion Engineering heamatopoietic stem cells with vIL-10 can protect cardiac allografts from acute rejection and prolong cardiac allografts survival.更多还原