Xu Ming-Qing, Yao Zhen-Xiang
Department of General Surgery, The first affiliated Hospital, Chongqing University of Medical Science, Chongqing, China.
World J Gastroenterol. 2003 Jan;9(1):141-7. doi: 10.3748/wjg.v9.i1.141.
To investigate functional change of dendritic cells (DCs) derived from allogeneic partial liver graft undergoing acute rejection in rats.
Allogeneic (SD rat to LEW rat) whole and 50 % partial liver transplantation were performed. DCs from liver grafts 0 hr and 4 days after transplantation were isolated and propagated in the presence of GM-CSF in vitro. Morphological characteristics of DCs propagated for 4 days and 10 days were observed by electron microscopy. Phenotypical features of DCs propagated for 10 days were analyzed by flow cytometry. Expression of IL-12 protein and IL-12 receptor mRNA in DCs propagated for 10 days was also measured by Western blotting and semiquantitative RT-PCR, respectively. Histological grading of rejection were determined.
Allogeneic whole liver grafts showed no features of rejection at day 4 after transplantation. In contrast, allogeneic partial liver grafts demonstrated moderate to severe rejection at day 4 after transplantation. DCs derived from allogeneic partial liver graft 4 days after transplantation exhibited typical morphological characteristics of DC after 4 days' culture in the presence of GM-CSF. DCs from allogeneic whole liver graft 0 hr and 4 days after transplantation did not exhibit typical morphological characteristics of DC until after 10 days' culture in the presence of GM-CSF. After 10 days' propagation in vitro, DCs derived from allogeneic whole liver graft exhibited features of immature DC, with absence of CD40, CD80 and CD86 surface expression, and low levels of IL-12 proteins (IL-12 p35 and IL-12 p40) and IL-12 receptor (IL-12Rbeta(1) and IL-12Rbeta(2)) mRNA, whereas DCs from allogeneic partial liver graft 4 days after transplantation displayed features of mature DC, with high levels of CD40, CD80 and CD86 surface expression, and as a consequence, higher expression of IL-12 proteins (IL-12 p35 and IL-12 p40) and IL-12 receptors (IL-12Rbeta1 and IL-12Rbeta2) mRNA than those of DCs both from partial liver graft 0 hr and whole liver graft 4 days after transplantation (P<0.001) was observed.
DCs derived from allogeneic partial liver graft undergoing acute rejection display features of mature DC.
研究大鼠同种异体部分肝移植急性排斥反应中移植肝来源树突状细胞(DC)的功能变化。
进行同种异体(SD大鼠到LEW大鼠)全肝和50%部分肝移植。分离移植后0小时和4天的肝移植组织中的DC,并在GM-CSF存在的条件下于体外扩增。通过电子显微镜观察培养4天和10天的DC的形态特征。通过流式细胞术分析培养10天的DC的表型特征。分别通过蛋白质免疫印迹法和半定量逆转录-聚合酶链反应检测培养10天的DC中IL-12蛋白表达及IL-12受体mRNA表达。确定排斥反应的组织学分级。
同种异体全肝移植在移植后第4天未表现出排斥反应特征。相反,同种异体部分肝移植在移植后第4天表现出中度至重度排斥反应。移植后4天的同种异体部分肝移植来源的DC在GM-CSF存在下培养4天后呈现出典型的DC形态特征。移植后0小时和4天的同种异体全肝移植来源的DC在GM-CSF存在下培养10天后才呈现出典型的DC形态特征。体外培养10天后,同种异体全肝移植来源的DC表现为未成熟DC特征,缺乏CD40、CD80和CD86表面表达,IL-12蛋白(IL-12 p35和IL-12 p40)及IL-12受体(IL-12Rβ(1)和IL-12Rβ(2))mRNA水平较低;而移植后4天的同种异体部分肝移植来源的DC表现为成熟DC特征,CD40、CD80和CD86表面表达水平较高,因此,其IL-12蛋白(IL-12 p35和IL-12 p40)及IL-12受体(IL-12Rβ1和IL-12Rβ2)mRNA表达高于移植后0小时的部分肝移植及移植后4天的全肝移植来源的DC(P<0.001)。
发生急性排斥反应的同种异体部分肝移植来源的DC表现为成熟DC特征。