摘要
AIM: To investigate functional change of dendritic cells (DCs)derived from allogeneic partial liver graft undergoing acuterejection in rats.METHODS: Allogeneic (SD rat to LEW rat) whole and 50 %partial liver transplantation were performed. DCs from livergrafts 0 hr and 4 days after transplantation were isolated andpropagated in the presence of GM-CSFin vitro. Morphologicalcharacteristics of DCs propagated for 4 days and 10 dayswere observed by electron rmicroscopy. Phenotypical featuresof DCs propagated for 10 days were analyzed by flowcytometry. Expression of IL-12 protein and IL-12 receptormRNA in DCs propagated for 10 days was also measured byWestern blotting and semiquantitative RT-PCR, respectively.Histological grading of rejection were determined.RESULTS: Allogeneic whole liver grafts showed no featuresof rejection at day 4 after transplantation. In contrast,allogeneic partial liver grafts demonstrated moderate tosevere rejection at day 4 after transplantation. DCs derivedfrom allogeneic partial liver graft 4 days after transplantationexhibited typical morphological characteristics of DC after 4days' culture in the presence of GM-CSF. DCs from allogeneicwhole liver graft 0 hr and 4 days after transplantation didnot exhibit typical morphological characteristics of DC untilafter 10 days' culture in the presence of GM-CSF. After 10days' propagationin vitro, DCs derived from allogeneic wholeliver graft exhibited features of immature DC, with absenceof CD40, CD80 and CD86 surface expression, and low levelsof IL-12 proteins (IL-12 p35 and IL-12 p40) and IL-12receptor (IL-12Rβ1 and IL-12Rβ2) mRNA, whereas DCs fromallogeneic partial liver graft 4 days after transplantationdisplayed 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-12Rβ1 and IL-12Rβ2) mRNA thanthose of DCs both from partial liver graft 0 hr and whole livergraft 4 days after transplantation (P＜0.001) was observed.CONCLUSION: DCs derived from allogeneic partial liver graftundergoing acute rejection display features of mature DC.
AIM:To investigate functional change of dendritic cells(DCs) derived from allogeneic partial liver graft undergoing acute rejection in rats. METHODS: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. RESULTS: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-12Rβ_1 and IL-12Rβ_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-12Rβ_1 and IL-12Rβ_2)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. CONCLUSION:DCs derived from allogeneic partial liver graft undergoing acute rejection display features of mature DC.
基金
the Medical Scientific Foundation of Chongqing City, No.01-2-029
