Th17细胞分化调控在干眼治疗中的研究现状

干眼是一种影响眼表的炎症性自身免疫疾病。近年来,致病效应辅助性T细胞17 (Th17)在干眼发病过程中的关键作用备受关注。Th17细胞通过产生IL-17来拮抗调节性T细胞(Treg),导致Th17细胞进一步扩增以及相关炎症因子的过度表达;在体内阻断IL-17可以减少Th17细胞扩增并促进Treg功能恢复,从而显著降低干眼的严重程度。因此,调控Th17细胞向Treg分化的抗炎药物可有效抑制干眼炎症,有望成为治疗干眼的新型抗炎药物。     

血清淀粉样蛋白A、高迁移率族蛋白1、CD64指数在白内障患者术后眼内炎诊断中的应用

目的　探讨血清淀粉样蛋白A（serum amyloid A protein,SAA）、高迁移率族蛋白1（high-mobility group box 1,HMGB1）、CD64指数与白内障术后眼内炎患者炎症因子间的相关性。方法　选择我院2015年1月至2019年11月白内障术后并发眼内炎患者为感染组（48例48眼）,未发生眼内炎患者为对照组（50例50眼）。绘制ROC曲线,分析两组患者血清SAA、HMGB1、CD64指数及白细胞介素1-β（IL-1β）、肿瘤坏死因子-α（TNF-α）及白细胞介素-17（IL-17）在诊断眼内炎中的价值。结果　感染组患者血清IL-1β、IL-17、TNF-α、SAA、HMGB1及CD64指数均显著高于对照组（均为P<0.05）。感染组中出现前房积脓患者血清SAA、HMGB1及CD64指数均显著高于未出现前房积脓者（均为P<0.05）。感染组患者血清SAA与其IL-1β及TNF-α,HMGB1与TNF-α,CD64指数与IL-1β、IL-17、TNF-α均呈明显正相关（均为P<0.05）。ROC曲线分析结果显示,血清IL-1β、IL-17及TNF-α在诊断白内障术后眼内炎中的效能均不高（均为AUC<0.750）;血清SAA、HMGB1及CD64指数在诊断白内障术后眼内炎中均有较高效能（均为AUC＞0.750）;各指标单独应用时,以CD64指数诊断效能最高［AUC=0.859,95%CI(0.756~0.963),灵敏度及特异度分别为86.4%和57.7%）］;SAA、HMGB1及CD64指数联合应用能有效提高各指标单独应用效果,其诊断效能最高［AUC=0.948,95%CI(0.892~1.000),灵敏度及特异度分别为90.9%和88.5%］。结论　血清SAA、HMGB1及CD64指数在诊断白内障术后眼内炎、反映眼内炎严重程度中具有一定的价值,有望成为临床预测眼内炎的新型指标。     

HSV-1上调白细胞介素-18 mRNA在小鼠角膜组织中表达的研究

目的研究单纯疱疹病毒Ⅰ型(HSV-1)感染小鼠眼球后,白细胞介素-18(IL-18)在角膜组织中的表达及IL-18的表达与单纯疱疹性角膜基质炎之间的关系。方法用106PFU的HSV-1感染BALB/c鼠的角膜后,在裂隙灯显微镜下观察角膜的临床变化,组织学检查角膜的病理改变;用RT-PCR和ELISA法检测IL-18在角膜组织中的表达水平。结果HSV-1引起的角膜基质炎在病毒感染后的第10d明显可见,在病毒感染后的第14~21d达到高峰。RT-PCR检测的数据显示:HSV-1感染的早期即可诱导IL-18mRNA在角膜组织中的表达,并且表达持续存在;IL-18mRNA的表达高峰时间(3~21d)位于临床疾病出现之前和临床疾病发展的过程中。ELISA检测角膜提取物中的IL-18蛋白显示了相似的结果。结论HSV-1上调IL-18在小鼠角膜组织中的表达;IL-18的表达与角膜基质炎的发生和发展密切相关;IL-18诱导Th1细胞介导的对单纯疱疹病毒特异性的免疫反应,导致单纯疱疹性角膜基质炎。     

植物凝集素诱导甲状腺相关眼病患者外周单核细胞与眼眶成纤维细胞共培养体系分泌IL-6、IL-17A的研究

背景 甲状腺相关眼病(TAO)的致病机制尚未明确,目前认为其是一种自身免疫性疾病,研究证实白细胞介素-17A(IL-17A)在多种自身免疫性疾病的发生和发展中起着重要作用,但其是否参与TAO的发病过程目前尚不明确. 目的 探讨培养外周血单个核细胞(PBMCs)与眼眶成纤维细胞(OFs)共培养体系中分泌的IL-17A是否参与TAO的发病过程及其可能的作用机制. 方法 采集2014年4-12月在中南大学湘雅医院确诊的12例TAO患者的外周血及眼眶结缔组织作为TAO组,并采集同期因先天性眼球萎缩行义眼台植入术的8例患者外周血及眼眶结缔组织作为对照组,采用密度梯度离心法提取所有受试者血中PBMCs,采用组织块培养法培养OFs.采用流式细胞仪检测PBMCs中T淋巴细胞纯度,分别用Giemsa染色法和免疫组织化学法鉴定培养的OFs.以1∶20的比例将OFs与PMBCs在U型96孔板中共培养以建立共培养体系,分别在各组共培养体系中加入0、1.0、2.5、5.0、10.0 μg/ml植物凝集素(PHA)刺激72 h,用ELISA法检测共培养体系上清液中IL-6、IL-17A质量浓度以及共培养体系细胞膜中总IL-17A受体(IL-17RA)的质量浓度.比较不同质量浓度PHA作用后各组共培养体系上清液中IL-6、IL-17A、IL-17RA质量浓度的差异. 结果 TAO组和对照组PBMCs中T淋巴细胞的纯度分别为(81.10±0.21)％和(80.05 ±0.38)％,组间差异无统计学意义(t=0.923,P＞0.05).体外培养的OFs对Vimentin呈阳性反应,Giemsa染色阳性,证实为成纤维细胞.1.0 μg/ml PHA作用后共培养体系中PBMCs和OFs增生能力最强,PHA质量浓度＞1.0μg/ml时,PBMCs出现凋亡,OFs增生明显减弱,相同质量浓度PHA作用下,TAO组的PBMCs和OFs增生均较对照组快.不同质量浓度PHA作用下TAO组与对照组共培养体系IL-6、IL-17A和IL-17RA质量浓度总体比较,差异均有统计学意义(IL-6:F分组=12.561,P=0.000;F质量浓度=23.356,P=0.001.IL-17A:F分组=12.037,P=0.000;F质量浓度=19.206,P=0.000.IL-17RA:F分组=16.216,P=0.000;F质量浓度=4.627,P=0.018).1.0μg/ml PHA作用后各组共培养体系上清液中IL-6、IL-17A和IL-17RA质量浓度均达峰值,随着PHA质量浓度的增加,IL-6、IL-17A和IL-17RA质量浓度均逐渐下降,差异均有统计学意义(均P＜0.05);相同质量浓度PHA作用下TAO组共培养体系上清液中IL-6、IL-17A、IL-17RA质量浓度明显高于对照组,差异均有统计学意义(均P＜0.05). 结论 1.0 μg/ml PHA刺激可增强共培养体系中PBMCs和OFs的增生及免疫炎性因子的分泌,IL-17A能够放大细胞免疫反应和炎症反应,从而参与TAO的发病过程.     

炎性细胞因子在蒸发过强型干眼的发病机制中的作用

目的：探讨炎性细胞因子在蒸发过强型干眼的发病机制中的作用。

方法：选择2010-01/2015-01我院诊治的蒸发过强型干眼患者39例65眼为研究对象。另取健康体检者30例53眼为对照组,对所有患者进行抗炎药物治疗。治疗前测定两组泪液TNF-α、IL-1、IL-6、IL-8水平进行组间比较,并于治疗后1、4、10wk检测试验组上述各炎性因子水平进行组内比较。

结果：对照组的泪液TNF-α、IL-1、IL-6、IL-8水平均显著低于试验组,差异具有统计学意义(P<0.05)。治疗后1、4、10wk时观察指标评价结果存在统计学意义(P<0.05),提示随着治疗时间的延长,患者各项观察指标均逐渐朝向正常值方向变化。治疗后1、4、10wk患者泪液TNF-α、IL-1、IL-6、IL-8比较存在统计学差异,上述各项指标均呈逐渐下降趋势,具有统计学差异(P<0.05)。

结论：TNF-α、IL-1、IL-6、IL-8在蒸发过强型干眼症患者中特异性升高,可作与疾病的发生发展密切相关,可作为临床诊断或疗效监测的重要参考指标。     

Vogt-Koyanagi-Harada综合征患者外周血中白细胞介素6及17的表达

白细胞介素6(IL-6)不仅通过直接与特异的受体结合发挥多种功能,而且可以诱导分泌IL-17的致病性辅助性Th17细胞的分化.它在类风湿性关节炎(RA)、系统性红斑狼疮(SLE)等自身免疫性疾病中发挥着重要作用[1].Vogt-KoyanagiHarada(VKH)综合征病因和发病机制尚不完全清楚,一般认为主要与自身免疫反应有关[2].而IL-6在VKH综合征中的作用目前尚不清楚.为了解IL-6在VKH综合征中的作用,我们应用酶联免疫吸附测定( ELISA)法检测了一组不同时期VKH综合征患者外周血中IL-6和IL-17的含量,并对其意义进行了探讨.现将结果报道如下.     

IL-17在单纯疱疹性角膜基质炎小鼠中表达的变化

目的:研究IL-17在小鼠单纯疱疹性角膜基质炎模型中的表达。方法:将1.0×106空斑单位的单纯疱疹病毒1型KOS毒株接种于BALB/c鼠的角膜上,建立HSK动物模型。免疫组织化学染色观察IL-17在角膜中的表达。分别取正常小鼠及接种病毒后的第1～3,7,10,14,21,28d,用毛细管取小鼠的左眼眼眶静脉窦血1mL,分离淋巴细胞,行荧光抗体染色,用流式细胞仪检测IL-17阳性的CD4+T细胞的表达。在裂隙灯显微镜下观察角膜的变化,检查角膜的组织学病理改变。结果:实验组中,角膜和外周血均有IL-17表达。实验组角膜基质内炎性细胞、角膜混浊程度非常严重。IL-17的表达与HSK小鼠炎症表现的严重程度成正相关(r=0.609,P<0.01)。结论:IL-17在HSK小鼠模型中呈高表达,且IL-17在HSK的发病过程中起一定作用。     

T淋巴细胞因子与单纯疱疹病毒性角膜炎的研究进展

单纯疱疹病毒性角膜炎(herpes simplex keratitis,HSK)是由病毒抗原引起的T淋巴细胞介导的迟发型变态反应。T淋巴细胞如何介导HSK免疫反应的详细机制目前仍不清楚,但从目前研究可总结:T淋巴细胞的细胞因子如IL-2,IL-10,IL-17,IFN-γ等促炎性因子在HSK的复发过程中起重要作用。本文将对T淋巴细胞及其细胞因子与HSK的研究进展做简要综述。     

辅助T细胞17在干眼症发病机制中的作用

干眼症是临床常见的眼表疾病, 患者出现明显的免疫稳态破坏。辅助T细胞亚群中产生白介素-17 (interleukin-17, IL-17)的辅助T细胞17(T helper type 17 cells, Th17)作为重要的免疫细胞广泛存在于人体中, 产生标志性的细胞因子如IL-17A、IL-17F和IL-22。转化生长因子-β、IL-6、IL-21和IL-23等可促进Th17细胞的分化。Th17细胞在干眼症的免疫反应中起着尤为重要的作用, 干眼症中Th17细胞相关细胞因子表达增高, Th17/调节性T细胞(regulatory T cell, Treg)比例上调, Th17细胞迁移至眼表可进一步导致干眼症的发生、发展。因此, 寻求Th17细胞相关的治疗靶点可能是治疗干眼症的关键因素。(国际眼科纵览, 2023, 47:62-67)     

辅助性T细胞17与葡萄膜炎关系的研究进展

辅助性T细胞(T helper cell,Th)17是一类近年来发现的CD4+Th细胞新亚型,由天然T细胞前体分化而来,具有独立的分化和调节机制,并大量产生白介素(interleukin,IL)-17、IL-6和肿瘤坏死因子-α,而不产生干扰素-γ和IL-4.Th17细胞在自身免疫性疾病以及感染性疾病的发生、发展、转归过程中有重要作用.在眼科与Behcet病、Vogt-小柳原田病等葡萄膜炎的发生及发展均相关.     

Investigating the relationship between serum interleukin-17 levels and systemic immune-mediated disease in patients with dry eye syndrome

Purpose

To investigate the association between dry eye syndrome (DE) and serum levels of interleukin (IL)-17 in patients with systemic immune-mediated diseases.

Methods

IL-17 and IL-23 levels were measured in the sera of patients whose tear production was <5 mm on the Schirmer test. Subjects included patients with chronic graft-versus-host disease (GVHD), rheumatoid arthritis (RA), Sjogren''s syndrome (SS), systemic lupus erythematosus (SLE), and no systemic disease. Corneal/conjunctival fluorescein staining was scored and the correlation between the score and the IL-17 level was evaluated.

Results

A strong correlation existed between IL-17 level and the type of systemic disease. IL-17 was significantly elevated in patients with chronic GVHD compared to those with RA and SS. IL-17 was not detectable in patients with SLE or in those without systemic disease. IL-23 was not detected in any of the subjects. IL-17 was significantly increased in patients with high fluorescein staining scores.

Conclusions

Our data suggest that IL-17 is involved in the pathogenesis of DE in patients with systemic immune-mediated diseases.     

炎性细胞因子与新生血管性眼病

新生血管性眼病是引起人类不可逆视力损害的主要病因.尽管眼内抗血管内皮生长因子药物治疗已成为临床中新生血管性眼病的重要疗法之一,但该疗法本身疗效不稳定且存在诸多的潜在风险和并发症.因此探寻更明确有效的治疗靶点是必要的.研究表明,肿瘤坏死因子-α、白细胞介素-1、白细胞介素-6和白细胞介素-18等炎性细胞因子与新生血管性眼病关系密切,本文就近些年来与新生血管性眼病相关的炎性细胞因子作一综述.     

Analysis of Th17-associated cytokines in tears of patients with dry eye syndrome

Purpose

To determine the levels of Th17-associated cytokines, particularly interleukin (IL)-17 and IL-22 in tears of patients with dry eye syndrome.

Methods

Tear samples were collected from 20 healthy volunteers, 20 dry eye (DE) patients with non-Sjögren''s syndrome (NSSDE) and 20 DE patients with Sjögren''s syndrome (SSDE). Symptom questionnaire was self-administered and multiple dry eye disease (DED)-related clinical tests were performed. The levels of IL-17 and IL-22 in tears were measured by enzyme-linked immunosorbent assay.

Results

The levels of IL-17 and IL-22 were significantly increased in tears of DE patients compared with those of controls and also higher in SSDE patients compared with those of NSSDE patients (P<0.05). Moreover, the levels of IL-17 and IL-22 were positively correlated with questionnaire score and keratopathy score but negatively correlated with tear film break-up time and Schirmer I test in both NSSDE and SSDE patients (P<0.05).

Conclusions

The levels of IL-17 and IL-22 in tears were significantly increased in DE patients, which were associated with the disease severity. Therefore, Th17 cell-associated cytokines, particularly IL-17 and IL-22, may have important roles in the immunopathogenesis of the DED.     

IL-17在实验性自身免疫性葡萄膜炎大鼠模型眼部的表达

目的研究IL-17阳性细胞在实验性自身免疫性葡萄膜炎（EAU）大鼠模型眼部的表达。方法用光感受器间维生素A类结合蛋白（IRBP）和氟氏完全佐剂等量混合注射于20只Lewis大鼠的右后足底部为实验组,于建模后第7、10、11、12、13、14、21、28天用裂隙灯显微镜进行眼前节检查,并根据de Smet的标准对炎症进行分级。用0.3 mL肝素（5 000 U/mL）注射至左心室以抗凝血,然后取眼球制作冰冻切片。免疫组织化学染色观察IL-17多克隆抗体在眼组织中的表达,5只正常Lewis大鼠作为对照。结果用IRBP免疫Lewis鼠后第10天,裂隙灯显微镜下可见眼前节炎症表现,免疫后第7、14、21、28天各组大鼠眼部炎症平均得分为0.4、3.2、1.6和0.2。实验组大鼠眼组织切片的苏木精-伊红染色可见炎性细胞浸润和光感受器细胞破坏。免疫组织化学染色发现,实验组虹膜、睫状体和视网膜部位可见IL-17蛋白表达,而正常组未见IL-17蛋白表达。第7、14、21、28天各组大鼠眼部组织切片染色的平均得分分别为0.6、4、2、0.4。EAU大鼠眼部炎症表现的严重程度与IL-17的表达量呈正相关（r=0.968,P〈0.01）。结论IL-17蛋白在Lewis大鼠EAU模型中呈高表达,表明IL-17阳性表达细胞在EAU的发病过程中起一定作用。     

Th17相关细胞因子在干眼患者眼表的表达及其相关性研究

背景 干眼的发病机制尚未明确.越来越多的证据表明干眼是免疫介导的炎症过程,辅助性T细胞17（Th17）相关细胞因子可能在干眼中发挥重要作用,但这些炎性因子的具体作用尚有待证实. 目的 分析干眼患者眼表组织中Th17相关细胞因子表达的差异. 方法 采用前瞻性队列研究设计,纳入2011-2012年在北京大学第三医院确诊的Sjogren综合征（SS）患者20例（SS组）、非SS干眼患者20例（非SS干眼组）和正常志愿者20例（正常对照组）.所有患者均为50岁以上的绝经后妇女,在填写知情同意书后依次进入研究队列.所有患者填写眼表疾病指数（OSDI）评分问卷进行主观症状评分,并进行干眼相关检查,包括泪膜破裂时间（BUT）测定、角膜荧光素染色评分、泪液分泌试验Ⅰ（SⅠt）.采用PCR-Array法测定3个组患者眼表Th17相关细胞因子含量,包括眼表组织中白细胞介素-17A（IL-17A）、IL-6、IL-8、IL-22、IL-23 mRNA的表达,并分析结膜上皮细胞中IL-17A与OSDI、BUT、角膜荧光素染色和SⅠt结果的相关性. 结果 SS组、非SS干眼组和正常对照组受检者的OSDI评分分别为50.00（33.50,66.50）、45.00（35.50,55.00）和3.00（0.00,5.00）,SⅠt值分别为2.50（1.00,4.00）、5.00（2.00,5.00）和15.50（10.00,18.50）,BUT值分别为2.00（1.00,4.00）、4.00（3.00,5.00）和10.00 （10.00,12.00）,角膜荧光素染色评分分别为8.50 （6.00,12.00）、5.50（4.00,7.00）和0.00（0.00,0.00）.3个组间受检者的OSDI评分、SⅠt、BUT及荧光素染色评分的总体比较差异均有统计学意义（χ^2=34.11、28.13、93.66、92.25,P＜0.01）,其中SS组患者的OSDI评分、荧光素染色评分均高于非SS干眼组患者,差异均有统计学意义（χ^2=28.72、88.43,P＜0.01）,而SⅠt、BUT值均低于非SS干眼组,差异均有统计学意义（χ^2=25.91、87.46,P＜0.01）;非SS干眼组患者的OSDI评分、荧光素染色评分均明显高于正常对照?     

Enhanced release of IL-6 and IL-8 into tears in various anterior segment eye diseases

OBJECTIVE: To determine the levels of interleukin 6 (IL-6) and interleukin 8 (IL-8/CXCL-8) in tears collected from the eyes of normal individuals and of patients with different irritative eye diseases, in order to acquire information on the immunological changes occurring during the early postoperative period following various forms of eye surgery, including penetrating keratoplasty (PKP). METHODS: IL-6 and IL-8 levels were measured with the aid of human ultrasensitive ELISA kits in the non-stimulated tears of patients in the early postoperative period following PKP or cataract operation, and of patients with acute bacterial conjunctivitis or with a corneal foreign body. The IL-6 and IL-8 concentrations, the total amounts released in a given time and the rates of their release were calculated. RESULTS: A significant increase in IL-6 release was observed in all patient groups compared with the normal controls (p < or = 0.003). The IL-8 release levels were significantly higher in the tears of all patient groups (p < or = 0.03), except for the cataract operation group, where the IL-8 release was not significantly higher (p = 0.053) than in the control samples. No significant differences in IL-6 or IL-8 release were observed when the various patient groups were compared with each other. CONCLUSION: The release of IL-6 and IL-8 into the tears is enhanced in various anterior segment eye diseases, and this may be used as an indicator of various inflammatory reactions in the early postoperative period.     

Constitutive and inflammatory mediator-regulated fractalkine expression in human ocular tissues and cultured cells

PURPOSE: Fractalkine (FKN) is a dual-adhesion molecule-chemokine that plays a role in inflammation but has not been explored in the eye. In the current study, constitutive expression of FKN was identified in human iris and retina, and its regulation by various cytokines in endothelial cells (ECs) and stromal cells from human iris, retina, and choroid was investigated. METHODS: Human iris and retina explants were evaluated for FKN mRNA and protein expression using RT-PCR and immunohistochemistry, respectively. Cultured ocular ECs and stromal cells were stimulated with various inflammatory mediators (endotoxin; TNFalpha; interferon-gamma; interleukin (IL)-1alpha, -4, -10, -13, -17, and -18; and/or CD40 ligand, or combinations thereof), with FKN mRNA being subsequently evaluated by cDNA array and/or RT-PCR and FKN protein by enzyme-linked immunoculture assay (ELICA) and/or by Western blot analysis. RESULTS: Iris and retina explants constitutively expressed FKN protein in microvascular ECs and also in several stromal cell types. Iris and retina both express FKN mRNA. TNFalpha upregulated FKN in iris explants. All ocular microvascular ECs and stromal cultures expressed low FKN mRNA and/or protein levels, which were variably upregulated by endotoxin, TNFalpha, interferon-gamma, IL-1alpha, and/or CD40 ligand, but not by IL-18. In ECs, the Th2 cytokines IL-4 and -13, but not IL-10, reduced TNFalpha-induced FKN protein. IL-17, usually considered proinflammatory, reduced TNFalpha-induced FKN protein in ocular ECs. CONCLUSIONS: FKN is expressed in various ocular tissues and cells. Inflammatory mediator modulation of ocular FKN expression suggests that this adhesive chemokine may play important roles in regulating leukocyte efflux in inflammatory eye diseases, such as anterior uveitis and retinochoroiditis.     

龙胆泻肝汤抑制葡萄膜炎大鼠Notch信号通路活化的作用

目的　探讨龙胆泻肝汤（Longdan Xiegan Decoction，LXD）对实验性自身免疫性葡萄膜炎（experimental autoimmune uveitis，EAU）大鼠Notch信号通路活化的抑制作用及其对辅助性T细胞17（T helper 17，Th17）和调节性T细胞（regulatory T cell，Treg）表达水平的影响。方法　随机将雌性Lewis大鼠分为正常对照（NC）组、EAU模型组、LXD干预组。EAU模型组和LXD干预组大鼠诱导EAU，免疫后LXD干预组大鼠每天给予LXD灌胃处理，EAU模型组大鼠给予等量生理盐水灌胃。免疫后12 d观察大鼠眼部炎症表现，取三组大鼠眼球进行病理切片，观察病理学变化；实时荧光定量PCR（quantitative polymerase chain reaction，QT-PCR）和酶联免疫吸附实验（enzyme-linked immunosorbent assay，ELISA）检测免疫后12 d三组大鼠脾脏、淋巴结及眼组织中Notch1、DLL4、白细胞介素10（interleukin 10,IL-10）和IL-17 mRNA及蛋白的表达；流式细胞仪检测三组大鼠各组织中Th17、Treg细胞的表达。结果　病理检查结果表明，LXD对EAU大鼠眼部组织结构有明显的保护作用。QT-PCR和ELISA检测结果发现，与NC组相比，LXD干预组大鼠脾脏、淋巴结和眼组织中Notch1、DLL4、IL-10、IL-17 mRNA和蛋白表达水平均升高，但除IL-10外，其他明显低于EAU模型组（均为P<0.05）；流式细胞仪检测结果发现，EAU模型组大鼠的各组织中Th17/Treg比例均高于NC组，经LXD干预后，Th17细胞表达水平下降，Treg表达水平升高，两者比例趋向平衡。结论　LXD可有效降低EAU大鼠脾脏、淋巴结、眼组织中Notch1、DLL4、IL-10和IL-17 mRNA和蛋白的表达水平，改善Th17/Treg细胞比例的平衡，从而有效减轻EAU大鼠的眼部炎症，保护眼部组织结构，调节全身及眼部的免疫状态。     

Alteration of tear cytokine balance by eye closure: analysis by multicytokine assay

Purpose To study the effects of eye closure on the balance of tear cytokines.Methods A total of 17 healthy volunteers with normal eyes were recruited. Tears (10 μl) were collected by capillary outflow at 09:00. The right eyes were closed with cotton bandages for 3 h, then tears were collected from each eye separately. The concentrations of inflammatory cytokines, interleukin (IL)-1β, IL-6, IL-8, IL-10, IL-12p70, and tumor necrosis factor (TNF)-α were measured by cytometric bead array.Results Although the concentration of each cytokine varied between eyes, the increasing ratio for 3 h showed a similar pattern in each cytokine. IL-1β, IL-6, IL-10, IL-12p70, and TNF-α increased and IL-8 decreased in normal eyes. After eye closure, the increase of IL-10 significantly diminished (P=0.037); however, the other cytokines were not affected.Conclusions Eye closure decreased the concentration of IL-10 in tears, which might cause a potentially pro-inflammatory status of the ocular surface.