Background T helper cell 17 (Th17),a newly discovered subset of CD4+ T cells,have been found to play an important role in dry eye disease in animal model.Further investigation should be done on the immunopathogenesis of Th17 cells in dry eye patients.Objective This study was to analyze the expression status of interleukin-17 (IL-17) and IL-22 in tear and supernatant of peripheral blood mononuclear cells (PBMCs) of dry eye patients and their correlation with clinical symptom and sign.Methods Twenty Sj(o)gren syndrome (SS)patients,twenty non-Sj(o)gren syndrome (NSS) patients were included in Wuxi Second Hospital from 2010 to 2011,and twenty healthy volunteers were recruited at the same period.All of subjects understood the purpose and procedure of research and written informed consent was obtained form each subject initial of this study.Dry eye symptom questionnairs were self-answered and multiple dry eye disease-related clinical tests,including the breakup time of tear film (BUT),Schirmer Ⅰ test (S Ⅰ t) and corneal fluorescein staining were performed.The periphery blood of 3 ml and tear were collected in all the subjects,and IL-17 and IL-22 levels in supernatant of PBMCs and tear were detected by enzyme-linked immunosorbent assay (ELISA).The correlations between levels of IL-17 and IL-22 with BUT,S Ⅰ t,corneal fluorescein staining and dry eye scores were analyzed.Results The dry eye scores reduced,BUT prolonged,S Ⅰ t increased and corneal fluorescein dye decreased from SS group,NSS group to normal control group,with significant differences among the three groups (dry eye scores:H =40.81,P＜0.01 ; BUT:H =40.15,P＜0.01 ;S Ⅰ t..H=50.07,P＜0.01 ;corneal dye scores:H=40.52,P＜0.01).The concentration of IL-17 in the supernatant of PBMCs in the SS patients,NSS patients and normal controls were (964.92±124.83)ng/L,(718.85± 115.89)ng/L and (341.95±85.08) ng/L,showing a statistically significant difference among them (F=162.95,P＜0.01).The levels of IL-17 in the tear were (440.69±126.09) ng/L,(364.33±126.85) ng/L and (61.16±11.60) ng/L in the SS group,NSS group and normal control group respectively,exhibiting an elevated level in the SS group and NSS group compared with the control group (F=75.27,P＜0.01).In addition,the levels of IL-22 in the supernatant of PBMCs in the SS patients,NSS patients and normal controls were (98.77± 11.27) ng/L,(79.65 ± 11.01) ng/L and (32.78±9.34) ng/L,and those in the tear were (22.22 ± 8.96) ng/L,(14.92 ±4.35) ng/L and (10.47 ± 2.67) ng/L,with significant differences among the three groups (F =206.27,P＜0.01 ;F =19.87,P＜0.01).The significant correlations were found between the IL-17 and IL-22 concentration in the supernatant of PBMCs and tear with corneal fluorescein staining scores and S Ⅰ t.Conclusions The contents of IL-17 and IL-22 in PBMCs and tear upregulate in the SS and NSS patients,indicating that Th17 plays a key role in the immunity mechanism of dry eye.%背景 辅助T细胞17(Th17)是一种新发现的CD4+T细胞亚群,动物研究显示,Th17细胞及其效应因子白细胞介素-17(IL-17)在干眼的发病和发展过程中发挥重要作用,但其发病机制还需进一步证实.目的 检测干眼患者泪液及外周血单核细胞(PBMCs)培养上清液中IL-17和IL-22的表达水平,探讨其与干眼临床症状及体征的相关关系. 方法 收集在无锡第二医院眼科确诊的Sj(o)gren综合征(SS)、非Sj(o)gren综合征(NSS)患者各20例,并纳入同期医院内职工和学生健康志愿者20人为正常对照组,3个组间患者年龄、性别比均匹配.所有患者行干眼症状调查量表评分、泪膜破裂时间(BUT)、泪液分泌试验Ⅰ(SⅠt)、角膜荧光素染色检查.收集所有受检者外周血3 ml,采用酶联免疫吸附(ELISA)法检测各组受检者PBMC培养上清液及泪液中IL-17、IL-22的质量浓度并进行比较,并与干眼临床症状和体征进行相关分析. 结果 NSS组、SS组及正常对照组干眼症状调查量表评分依次降低,SS组、NSS组及正常对照组BUT值依次延长,SⅠt值依次增大,角膜荧光素染色评分明显降低,3个组间上述指标的总体比较差异均有统计学意义(症状评分:H=40.81,P＜0.01；BUT:H=40.15,P＜0.01；S Ⅰ t:H=50.07,P＜0.01；角膜荧光素染色评分:H=40.52,P＜0.01).SS组、NSS组及正常对照组PBMCs培养上清液中IL-17质量浓度分别为(964.92±124.83)、(718.85±115.89)、(341.95±85.08) ng/L；泪液中IL-17质量浓度分别为(440.69±126.09)、(364.33±126.85)、(61.16±11.60) ng/L,SS组、NSS组患者PBMCs中和泪液中IL-17质量浓度明显高于正常对照组,3个组间总体比较差异均有统计学意义(F=162.95,P＜0.01；F=75.27,P＜0.01).SS组、NSS组及正常对照组PBMCs培养上清液中IL-22质量浓度分别为(98.77±11.27)、(79.65±11.01)、(32.78±9.34) ng/L,泪液中分别为(22.22±8.96)、(14.92±4.35)、(10.47±2.67) ng/L,表明SS组、NSS组患者PBMCs中和泪液中IL-22质量浓度明显高于正常对照组,3个组间总体比较差异均有统计学意义(F=206.27,P＜0.01；F=19.87,P＜0.01).干眼患者PBMCs培养上清液及泪液中IL-17、IL-22水平与角膜荧光素染色评分、SⅠt有明显关联,但与干眼症状量表评分、BUT均无相关性.结论 干眼患者PBMCs及泪液中IL-17、IL-22的质量浓度明显升高,进一步提示Th17在干眼免疫机制中发挥重要作用.
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