Effects On Recombinant Human Interleukin-10 Of Treg Cells And IL-10,TGF-β In Transplantation Of The Rabbit Skin

Objective:Observed the rejection time and the survival time of grafted skin, observed the changes of Treg cells,IL-10 and TGF-βin peripheral blood after rabbits'skin transplantation which treat with rhIL-10 or CsA.Discuss the changes of Treg cells,IL-10 and TGF-(3.To investigate the role of IL-10 in immunological rejection mechanism. Observed the effect of IL-10 or CsA or combined application,providing experimental evidence for reduce the CsA's side effects and searching new drugs.Materials:(1)36 rabbits were divided into two groups which were receptors and donors.Each donor rabbit preserved skin for one receptor,skin transplants on the back.Receptors were randomly divided into six groups:Group①:rhIL-10 5μg/kg/d; Group②:rhIL-10 10μg/kg/d; Group③:CsA 5mg/kg/d; Group④:CsA 10g/kg/d; Group⑤:rhIL-10 5μg/kg/d+CsA 5mg/kg/d; Group⑥:NS 1 ml/d。All groups were intramuscular injected durgs ten days started from one day before the surgery of skin transplantation. (2)Observed the situation of grafted skin after the skin transplantion everyday,to judge graft rejection with color darken,skin harden,red swelling,blisters and skin upwrap,to judge skin necrosis with the grafted skin more than 80% had appeared the phenomenon.Record the rejection time and the survival time of the grafted skin. (3)Each groud of rabbits'peripheral blood were collected at 1 day before surgey and 1thday,4thday,7thday,14thday,21thday,28thday after the operation.All the samples are analyzed by flow cytometry(FCM) and enzyme-linked immunosobent assay(ELISA), Record the ratio of CD4+CD25+Treg/ CD4+T cells and the concentration of IL-10,TGF-β.Statistical analysis has been performed with SPSS15.0. (4)The peripheral blood of rallits were drawn at the time of the 30th day after aoperation,detected IL-10 antibody in serum.Results:(1)The rejection time of experimental group are longer than control group.Except group①,other experimental groups compared with group⑥have significant differences(P<0.05). The survival time of experimental group are longer than control group and have singnificant differences(P<0.05).The survival time of group⑤are longer than group①, have singnificant differences (P＜0.05). group⑤are longer than group③,but have not singnificant differences (P>0.05).(2)The proportion of CD4+CD25+ regulatory T cells in periperal blood have increased after the skin transplantation.4thday,7thday after the operation,group①and group②are hi gher than group⑥,group③and group④, have significant differences(P<0.05). group③a nd group④are lower than control group and have significant differences(P<0.05).(3) The concentration of IL-10 in periperal blood has significant increased after the skin transplantation(P<0.05). lthday,4thday,7thday after the operation, group①and g roup②ar e higher than group⑥,group③and group④(P<0.05).(4) The concentration of TGF-βin periperal blood has significant increased after the skin transplantation (P<0.05).There was not obvious statistic difference between group①,group②,group③,group④and g roup⑤after skin transplantation(P>0.05). (5)The concentration of IL-10 is positive correlation with the proportion of CD4+CD25+ regulatory T cells.(6)There was negative detection of IL-10 antibody in alls rabbits'blood.Conclusion:(1) IL-10 can delay the rejection time on rabbits'skin transplantation,and can prolong the survival time.(2)There is synergistic effect between IL-10 and CsA.(3)The role of IL-10 inhibit allograft rejection may be related to CD4+CD25+ regulatory T cells and IL-10,and may be independent with TGF-β.(4)There is not IL-10 antibody in rabbits'blood after intramuscular injected rhIL-10.