重组hIL-10抗家兔皮肤移植排斥反应中血清IL-10及其抗体的检测

目的检测重组人白细胞介素-10（recombinant human interleukiu-10,rhIL-10）作用后皮肤移植家兔血清中IL-10及其抗体的变化,探讨外源性hIL-10诱生IL-10增强抑制效应和外源性hIL-10有无免疫原性,为rhIL-10的应用提供实验依据。方法家兔分为6组：①组rhIL-10低剂量,②组rhIL-10高剂量,③组环孢菌素（CsA）低剂量,④组CsA高剂量,⑤组rhIL-10＋CsA低剂量联合,⑥组生理盐水组。移植术前1d开始用药,连续用药10d,肌肉注射。术前1d、术后1d、4d、7d、14d、21d、28d分别取外周血,分离血清,以ELISA方法检测血清中IL-10及IL-10抗体水平。结果①移植家兔血清IL-10检测,术后各组结果较术前均有升高,在术后7d达到高峰,术后4d、术后7d rhIL-10低剂量及高剂量组IL-10水平高于其余各组（P〈0.05）。CsA低剂量及高剂量组IL-10水平低于rhIL-10组及联合组（P〈0.05）。②移植家兔血清抗IL-10抗体检测,术后各组检测结果较术前均无明显升高（P〉0.05）,与抗体阳性对照比较有显著差异（P〈0.05）。结论 rhIL-10抗家兔皮肤移植反应中,家兔血清IL-10含量在皮肤移植后明显升高,表明外源IL-10诱导内源性IL-10分泌,从而增强抑制排斥反应;外源IL-10注射后家兔血清中没有检测出特异性抗体,外源性rhIL-10进入机体没有免疫原性。

Evaluation of IL-10 and IL-10 antibody in the reaction of rhIL-10 resisting the rabbits skin graft rejection

Objective To explore the mechanism of recombinant human interleukin-10 resisting the immunological rejection and provide laboratory evidence for the recombinant human interleukin-10＇s clinic utilization by observing the changes of IL-10 and anti-IL-10 antibody in the serum of rabbits treated with recombinant human interleukin-10 after skin graft.Methods The rabbits were divided into six groups,① rhIL-10 low dose group,② rhIL-10 high dose group,③ CsA low dose group,④ CsA high dose group,⑤ rhIL-10 and CsA low dose joint group,⑥ normal saline group.The medicine was continuously given by intramscular injection to the rabbits which were skin graft from the preoperative one day to the tenth day.The serum was separat from the skin grafting rabbits＇ blood on the preoperative one day,the first day,the fourth day,the seventh day,the fourteenth day,the twenty first day and the twenty eighth day after skin graft.Then test IL-10 and IL-10 antibody by ELISA assay.Results ①The results of IL-10 test show that：the serum IL-10 increases after skin graft compared with preoperative and reaches the peak concentration on the seventh day.On the fourth and the seventh day after transplation,group ① and ② was higher than the other groups（P0.05）.Moreover group ③ and ④ was lower than group ⑤（P0.05）.②The results of anti IL-10 antibody test show that：no significant difference in each two groups.Conclusion In the research of rhIL-10 resisting the rabbits skin graft,IL-10 in the serum increased obviously after the rabbits skin graft,which showed that more IL-10 of endogenesis could be induced to secrete with IL-10 of exogenous and then enchanced the level of the reaction of resisting the skin graft rejection.No anti-IL-10 antibody had been detected in the serum after the rabbits were treated with recombinant human interleukin-10,this provide an evidence that rhIL-10 of exogenous has no immunogenicity to the body.