| BACKGROUND: Allograft rejection remains a major obstacle to successful transplantation. The polymorphisms of histocompatibility antigens and the shortage of donor organs ensure that better matching is not a solution to the problem of immunologic barriers.Immunosuppressive agents inhibit all T cell or subset of T cells and cause nonspecific suppression of the immune system.To localize immunosuppressive signals to the region(s) of antigen presentation and recognition and to modulate the immune system at the level of antigen-specific lymphocyte priming and activation is a good method to avoid systemic immunosuppression. The ability to express genes with potential immunoregulatory capacity could reduce the immunogenicity of allografts and result in long-term graft survival.There have been conflicting reports of the influence of exogenous mammalian interleukin(IL)-10 on immune reactivity.In this study,AIM In part I To examine the feasibility of transferring interleukin(IL)-10 gene into rat hearts by directly injecting pSV-SPORT1-IL-10 into allograft hearts before transplantation and immune effect.In part Ⅱ TO investigate the effect of IL-10 on CD4+T cell polarized to Th1/Th2 in vitro. In part Ⅲ To investigate the transfer and expression of pSV-SPORT1-IL-10 in allograft heart by directly injected into allograft heart left ventrical wall befor transplantation.METHODS In part I Rat heart transplants(Wister→SD) were performed in five groups: groupl:no treatment, group2: pSV-SPORT1-IL-10, group3: pSV-10+CsA, group4:CsA,group5:IL-10.Allograft survival was determined bypalpating heartbeats.Test the serum IL-10 of recipients.Allograft tissues were also submitted for histological study. In part II After isolated the PBMCs and blood plasma from adult rat peripheral blood by Ficoll-Hypaque centrifugation, the PBMC culture procedure with or without the self-blood-plasma was applied to polarized T cells in vitro,these cells were polarized by PHA(20mg/L), non-PHA respectively.The polarizd rates of Th cell after 72h were estimated respectively by flow cytometry following two-color immunofluorescent staining. In partlll pSV-SPORTl-IL-10 was diretly injected into allograft heart befor transplantation. Seven days postoperation hemospasia from the recipients tail vein and check serum IL-10. Kill the receptors and do allograft heart muscle DNA electrophoretic analysis and immunohistochemical analysis. RESULTS In part I IL-10 expression was shown in heart allografts. Animals transfected with IL-10 showed prolongation of graft survival(21.00 + 3.34dvs8.71 ± 1.1 ld,t=l 1.43 p<0.001)compared to controls. Animals transfected with both IL-10 and CsA demonstrated a synergistic prolongation of allograft survival compared with animals treated with CsA or IL-10 treatment alone(38.14± 3.34dvsl9.57±1.51d,p<0.001;38.14±3.34dvs21.00±2.16d,p<0.001)). All of the serum of recipients have no IL-lO.In partHCD4+T cell would polarize to Thl/Th2 two subsets after self-cytokines and PHA activation in vitro.The polarized rates of T cell after cultured for 72h were group A: 2.81%/2.86% , group B: 3.13±0.60%/3.50 ±0.49%, group C: 17.80±0.88%/17.31±0.98% ,group D: 13.27±0.42%/21.68± 0.77% respectively. Group C vs group B ^1=43.15 Pthi<0.001, tth2 =39.46 Pth2 <0.001;Group A> B and C Thl:Th2=l, Group D vs group C Ui = 14.61 P,hi<0.001, ^2=11.21 Pth2 O.001 ; Group D ThlvsTh2 t=30.04 P<0.001. Group D polarized to Th2 severely. In partlll Histological study showed that allografts from untreated controls exhibited extensive lymphocyte infiltrating with loss of graft architecture by day 7 post transplantation while those from the IL-10 group showed less lymphocyte infiltrating and graft architecture was good.No serum IL-10 of the receptors ,aim gene expessed and exogenous was seen.CONCLUSION In part I Delivering IL-10 into donor hearts by directly injecting pSV-SPORTl-IL-10 into...
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