大鼠自身免疫性葡萄膜视网膜炎辅助T细胞的类细胞因子水平表达

Helper T cell cytokines expression in autoimmune uveoretinitis in rats

目的观察实验性自身免疫性葡萄膜视网膜炎(experimental autoimmune uveoretinitis,EAU)大鼠血清以及脾细胞培养上清液中辅助T细胞1/辅助T细胞2(helper T cell 1/helper T cell 2,Th1/Th2)类细胞因子水平。方法用Fmoc法合成光感受器间维生素A类结合蛋白R16多肽片段,联合免疫佐剂诱导EAU动物模型。在EAU高峰期取大鼠血清,分离脾细胞,培养后取上清液,应用酶联免疫吸附法检测血清以及脾细胞培养上清液中Th1类细胞因子(interferon-γ,IFN-γ、interleukin-2,IL-2)和Th2类细胞因子(IL-4、IL-10)水平。结果EAU组大鼠血清中IFN-γ和IL-2的浓度分别为(33.8±5.2)μg/L,(52.5±7.9)μg/L,显著高于空白对照组[(6.2±1.4)μg/L,(3.7±0.8)μg/L]和弗氏完全佐剂对照组[(complete Freund′s adjuvant,CFA);(9.2±1.9)μg/L,(5.1±1.1)μg/L](P<0.05);而IL-4、IL-10的浓度与空白对照组和CFA组相比差异无统计学意义。EAU组大鼠脾细胞培养上清液中IFN-γ、IL-2的浓度分别为(1 105.3±197.5)μg/L,(45.0±16.2)μg/L,显著高于空白对照组(5.2±1.7)μg/L,(4.1±1.3)μg/L和CFA组(25.1±5.9)μg/L,(5.1±1.9)μg/L(P<0.01),IL-4、IL-10的浓度与空白对照组和CFA组相比差异无统计学意义;CFA组大鼠脾细胞培养上清液中IFN-γ的浓度明显高于空白对照组(P<0.05),IL-2、IL-4、IL-10的浓度与空白对照组相比差异无统计学意义。结论在EAU高峰期,Th1类细胞因子水平显著升高。

Objective To investigate the expression of helper T cell 1/helper T cell 2（Th1/Th2） cytokines in serum and culture supernatants of splenic cells in rats with experimental autoimmune uveoretinitis （EAU）. Methods Fmoc procedure was used to synthesize the interphotoreceptor retinoid-binding protein （IRBP） R16 peptide in order to establish EAU animal model combined with immune adjuvant. The expression of interferon-γ（ IFN-γ）, interleukin-2 （IL-2）, IL-4 and IL-10 in serum, culture supernatant of splenic cells was determined by enzyme-linked immunosorbent assay kits. Results The serum levels of IFN-γ and IL-2 in EAU rats were （33.8 ± 5.2） μg/L and （52.5 ± 7.9 ） μg/L respectively, which were significantly higher than those in the control group [ （6.2 ± 1.4 ） μg/L, （3.7 ±0.8） μg/L] and complete Freund＇s adjuvant（CFA） group [ （9.2 ± 1.9 ） μg/L, （5.1 ± 1.1 ） μg/L] （P 〈0.05）. The serum levels of IL-4 and IL-10 in EAU rats showed no significant differences compared with the control or CFA group. The levels of IFN-3, and IL-2 in the culture supernatant of splenic cells in EAU rats were （ 1 105.3 ± 197.5 ） μg/L and （45.0 ± 16.2） μg/L, which were significantly different from those in the control group[ （5.2 ± 1.7） μg/L, （4.1 ± 1.3 ）μg/L] ） and CFA group [ （25.1 ±5.9 ） μg/L, （ 5.1 ± 1.9） μg/L] ] （P 〈0.01 ）. The levels of IL-4 and IL-10 were not significandy different from those in the control and CFA group. IFN-γlevel in the culture supernatant of splenic cells in CFA rats was significantly higher than those in the control group （P 〈 0.05 ） while the levels of IL-2, IL-4, IL-10 were not significantly different between the two groups. Conclusions An increased production of Th1 cytokine, IFN-γ and IL-2 was observed in the EAU rats, suggesting that EAU may be mainly induced by Th1 cells. （ Chin J Ophthalmol and Otorhinolaryngo1,2008,8：280-282）