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疾病状态下泪液中半乳糖凝集素-3的检测以及人角膜和结膜上皮的免疫组织化学和凝集素组织化学分析

Detection of galectin-3 in tear fluid at disease states and immunohistochemical and lectin histochemical analysis in human corneal and conjunctival epithelium.

作者信息

Hrdlicková-Cela E, Plzák J, Smetana K, Mĕlková Z, Kaltner H, Filipec M, Liu F T, Gabius H J

机构信息

1st Faculty of Medicine, Department of Ophthalmology, Charles University, U nemocnice 3, 128 00 Prague 2, Czech Republic.

出版信息

Br J Ophthalmol. 2001 Nov;85(11):1336-40. doi: 10.1136/bjo.85.11.1336.

Abstract

BACKGROUND/AIM: Components of the tear fluid contribute to the biochemical defence system of the eye. To reveal whether the immune mediator and lipopolysaccharide binding galectin-3 is present in tears, tear samples were collected from eyes in healthy and pathological states. Investigation of expression of galectin-3 and galectin-3 reactive glycoligands in normal human conjunctival and corneal epithelia was also initiated as a step to understand the role of galectin-3 in ocular surface pathology.

METHODS

Immunoblot analysis using either a rabbit polyclonal or a mouse monoclonal antibody against galectin-3 was employed to detect galectin-3 in tear fluid. Galectin-3 expression in tissue specimens was detected by immunocytochemistry employing A1D6 mouse monoclonal antibody, and galectin-3 reactive glycoligands were visualised by lectin histochemistry using labelled galectin-3.

RESULTS

Galectin-3 was found only in tears from patients with ocular surface disorders. It was expressed in normal corneal and conjunctival epithelia but not in lacrimal glands. Inflammatory leucocytes and goblet cells found in galectin-3 containing tear fluid also expressed galectin-3. Galectin-3 binding sites were detected on the surface of conjunctival and corneal epithelial cells co-localising with desmoglein.

CONCLUSIONS

This study revealed expression of galectin-3 in tear fluid obtained from patients with eye diseases. The role of this endogenous lectin (produced by inflammatory as well as epithelial cells) in antimicrobial action and inflammation modulation could be expected.

摘要

背景/目的:泪液成分有助于眼部的生化防御系统。为了揭示免疫介质和脂多糖结合蛋白半乳糖凝集素-3是否存在于泪液中,我们从健康和病理状态的眼睛中收集了泪液样本。作为了解半乳糖凝集素-3在眼表病理中作用的一个步骤,我们还对正常人结膜和角膜上皮中半乳糖凝集素-3及其反应性糖配体的表达进行了研究。

方法

使用抗半乳糖凝集素-3的兔多克隆抗体或小鼠单克隆抗体进行免疫印迹分析,以检测泪液中的半乳糖凝集素-3。使用A1D6小鼠单克隆抗体通过免疫细胞化学检测组织标本中的半乳糖凝集素-3表达,并使用标记的半乳糖凝集素-3通过凝集素组织化学观察半乳糖凝集素-3反应性糖配体。

结果

仅在眼表疾病患者的泪液中发现了半乳糖凝集素-3。它在正常角膜和结膜上皮中表达,但在泪腺中不表达。在含有半乳糖凝集素-3的泪液中发现的炎性白细胞和杯状细胞也表达半乳糖凝集素-3。在结膜和角膜上皮细胞表面检测到与桥粒芯糖蛋白共定位的半乳糖凝集素-3结合位点。

结论

本研究揭示了眼病患者泪液中半乳糖凝集素-3的表达。可以预期这种内源性凝集素(由炎症细胞和上皮细胞产生)在抗菌作用和炎症调节中的作用。

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