Okada A A, Sakai J, Usui M, Mizuguchi J
Department of Ophthalmology, Tokyo Medical College Hospital, Japan.
Ocul Immunol Inflamm. 1998 Jun;6(2):111-20. doi: 10.1076/ocii.6.2.111.4046.
Elevations of inflammatory cytokines have been implicated in the pathogenesis of experimental autoimmune uveoretinitis (EAU) in rats, although such analysis has relied on indirect methods of assessment such as measurement of mRNA content. In this study, we examined the feasibility of directly measuring cytokine concentrations in intraocular extracts prepared by ultrasonic disruption. Cytokines were measured by ELISA in eyes from EAU-induced Lewis rats immunized with interphotoreceptor retinoid-binding protein (IRBP), and compared to eyes from rats immunized with adjuvant only and from normal rats. Interferon-gamma (IFN-gamma), tumor necrosis factor-alpha (TNF-alpha), interleukin-2 (IL-2), interleukin-4 (IL-4), and interleukin-10 (IL-10) were detectable in EAU eyes at near peak inflammation, with IFN-gamma achieving the highest mean concentration (331 pg/ml). In eyes from rats immunized with adjuvant only and in normal eyes, IFN-gamma, TNF-alpha, and IL-10 were nondetectable. IL-2 and IL-4 were detected at significantly lower mean concentrations (32.3 pg/ml and 69.4 pg/ml, respectively) compared to EAU eyes (217 pg/ml and 230 pg/ml, respectively); IL-4 was also detected in eyes from rats immunized with adjuvant alone (141 pg/ml). Thus, a direct method of measuring intraocular cytokine concentrations was successfully applied to reveal an elevation of IFN-gamma, TNF-alpha, IL-2, IL-4, and IL-10 in EAU eyes from rats immunized with IRBP, compared to rats immunized with adjuvant alone and to normal rats. These cytokine elevations reflect the local intraocular environment near peak inflammation, and suggest an important role for these cytokines in the mechanisms of onset and resolution of EAU in rats.
炎症细胞因子的升高与大鼠实验性自身免疫性葡萄膜视网膜炎（EAU）的发病机制有关，尽管此类分析依赖于间接评估方法，如测量mRNA含量。在本研究中，我们检测了通过超声破碎制备的眼内提取物中直接测量细胞因子浓度的可行性。通过酶联免疫吸附测定法（ELISA）测量用视网膜色素上皮结合蛋白（IRBP）免疫诱导EAU的Lewis大鼠眼中的细胞因子，并与仅用佐剂免疫的大鼠和正常大鼠的眼睛进行比较。在炎症接近高峰时，EAU大鼠眼中可检测到干扰素-γ（IFN-γ）、肿瘤坏死因子-α（TNF-α）、白细胞介素-2（IL-2）、白细胞介素-4（IL-4）和白细胞介素-10（IL-10），其中IFN-γ的平均浓度最高（331 pg/ml）。在仅用佐剂免疫的大鼠眼中和正常眼中，未检测到IFN-γ、TNF-α和IL-10。与EAU大鼠眼（分别为217 pg/ml和230 pg/ml）相比，IL-2和IL-4的平均浓度显著较低（分别为32.3 pg/ml和69.4 pg/ml）；在仅用佐剂免疫的大鼠眼中也检测到IL-4（141 pg/ml）。因此，一种直接测量眼内细胞因子浓度的方法成功应用于揭示，与仅用佐剂免疫的大鼠和正常大鼠相比，用IRBP免疫的大鼠EAU眼中IFN-γ、TNF-α、IL-2、IL-4和IL-10升高。这些细胞因子的升高反映了炎症接近高峰时的局部眼内环境，并表明这些细胞因子在大鼠EAU的发病和消退机制中起重要作用。
