IL-2-PE66抑制大鼠角膜内皮细胞MHC抗原表达的研究

Effect of IL-2-PE66 on MHC antigen expression in corneal endothelium cells of rat

目的了解正常大鼠角膜内皮细胞在体外经γ-干扰素诱导后,组织相容性复合体(MHC)-Ⅰ、Ⅱ类抗原异常表达的情况,并观察比较IL-2-PE66、环孢素A(CsA)对这种异常表达的免疫抑制作用。方法采用ACAS-570黏附式细胞分析仪和免疫荧光技术,对体外原代培养经γ-干扰素诱导的大鼠角膜内皮细胞进行MHC-Ⅰ、Ⅱ类抗原表达量的测定,并观察加入白细胞介素-2(IL-2)-PE66、CsA后,MHC-Ⅰ、Ⅱ类抗原表达量的变化。结果γ-干扰素能明显诱导大鼠角膜内皮细胞MHC-Ⅰ、Ⅱ类抗原异常表达,而IL-2-PE66、CsA能明显抑制这种异常表达,二者之间的免疫抑制效果无明显差异。结论IL-2-PE66及CsA均能明显抑制大鼠角膜内皮细胞在体外经γ-干扰素诱导后,MHC-Ⅰ、Ⅱ类抗原的异常表达。提示IL-2-PE66是一种高特异性的强效免疫毒素,是未来抗角膜移植排斥的新型基因药物。

Objective Corneal transplantation is the most common type of clinical grafting performed in humans and it shows a high incidence of allograft rejection in ＂high risk ＂eyes, so prevention and treatment of rejection is the most important topic in corneal transplantation. This study was to investigate the abnormal expression of major histocompatibility complex（MHC） antigen-Ⅰ , Ⅱ induced by interferon gamma （INF-γ） in vitro in corneal endothelium cells of rat,and observe and compare the inhibiting effects of interleukin-2-pseudomonas exotoxin66（IL-2-PE66） and CsA on the expression of MHC antigen. Methods The rat corneal endothelium cells primarily cultured in vitro were divided into two groups,in which IL-2-PE66 and CsA treatments were applied and another one without additional treatment except INF-γ application. These two groups were subdivided into a,b, c,d groups. 500 × 10^3 U/L γ-INF was added in a,b,c groups and then 70 mg/L IL-2-PE66,300 g/L CsA was added into a,b,c group separately,and d group was as negative control. The expression of MHC antigen-Ⅰ , Ⅱ was detected before and after addition of IL-2-PE66,300 g/L CsA. Results Before addition of INF-γ,the expressive of M HC antigen- Ⅰ was 97.8 ± 8. 1 and antigen- Ⅱ was 0. After induction by INF-γ,the expressive amount of MHC antigen- Ⅰ was 1 006.3 ± 13.2 and that of antigen- Ⅱ was 406.5 ± 10. 5 ,indicating a significant difference in MHC antigen-Ⅰ , Ⅱ levels before and after induced by INF-γ （P 〈 0. 05 ）. The expressive amount of MHC antigen- Ⅰ , Ⅱ was 598.2 ± 12.5,197.5 ±6.8 in group a respectively,and those of group b were 609.5 ± 12.9 and 198.5 ±6.9. A significant difference was seen in expression of MHC antigen-Ⅰ , Ⅱ between group a and group c or group b and group c （ P 〈 0.05 ） , but there was no statistically difference between group a and group b （ P 〉 0. 05 ）. Conclusion INF-γ can induce the abnormal expression of MHC antigen- Ⅰ and Ⅱ in corneal endothelium cells of rats in vitro,and IL-2-PE66 and CsA can inhibit the antigen expression to different degree.