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在进行非协调性异种器官移植时,目前已经证实表达人α-1,2-岩藻糖苷转移酶(HT)或者补体调节蛋白的供体动物器官均可以部分克服超急性排斥反应.本文的研究目的是探讨是否共表达人HT和补体调节蛋白(衰变加速因子与CD59)的转基因小鼠的外周血单核细胞能更有效的克服异种移植排斥反应.利用受精卵显微注射技术建立转人HT,DAF和/或CD59基因的小鼠动物模型,流式细胞技术筛选表达不同基因的转基因小鼠.将小鼠的外周血单核细胞与15%的人血清共孵育,检测细胞表面天然抗体的沉积、补体的激活以及黏附分子的表达等.三种目的基因均可以在转基因小鼠体内表达,并且HT基因的表达显著减少了引起异种移植排斥反应的主要抗原半乳糖α-1,3-半乳糖(α-Gal)的表达.功能实验表明,与单一目的基因表达的转基因小鼠相比,共表达HT/DAF或HT/CD59的转基因小鼠的外周血单核细胞对抗人血清介导溶破细胞的能力显著增强.而且三基因共表达的外周血单核细胞对抗人血清介导溶破细胞的能力最强,可以克服超急性排斥反应并且可以减少黏附分子的表达.高水平表达人HT,DAF和CD59基因的转基因小鼠可以完全克服异种器官移植超急性排斥反应并且可以部分克服急性血管排斥反应.本研究表明,表达三种基因的转基因小鼠的器官异种移植时存活时间可以显著延长,对异种移植来说也许是更合适的选择.
In the case of non-coordinating xenotransplantation, donor animal organs that express human α-fucosyltransferase (HT) or complement regulatory proteins have now been shown to partially overcome hyperacute rejection. In this study The aim is to investigate whether peripheral blood mononuclear cells from transgenic mice that co-express human HT and complement regulatory proteins (decay accelerating factor and CD59) are more effective in overcoming xenograft rejection. , DAF and / or CD59 gene, and the transgenic mice expressing different genes were screened by flow cytometry.The mouse peripheral blood mononuclear cells were co-incubated with 15% human serum to detect the expression of native antibody Deposition, activation of complement, expression of adhesion molecules, etc. All three target genes can be expressed in transgenic mice, and the expression of HT gene significantly reduces the expression of the major antigen galactose α-1,3-hemoglobin which causes xenograft rejection Lactose (α-Gal) .Functional experiments showed that compared with transgenic mice expressing a single gene of interest, the peripheral blood mononuclear cells of transgenic mice that co-expressed HT / DAF or HT / CD59 Anti-human serum-mediated ability to dissolve cells was significantly enhanced.And the three gene co-expression of peripheral blood mononuclear cells against human serum-mediated ability to solve the most powerful cells can overcome hyperacute rejection and can reduce the expression of adhesion molecules Transgenic mice with high levels of human HT, DAF and CD59 genes can completely overcome hyperacute rejection of xenotransplantation and partially overcome acute vascular rejection.The present study shows that organ transplantation of transgenic mice expressing three genes When the survival time can be significantly extended for the xenotransplantation may be a more appropriate choice.