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

机译:疾病状态下泪液中半乳凝集素3的检测以及人角膜和结膜上皮的免疫组化和凝集素组织化学分析

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摘要

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.

机译:背景/目的-泪液的成分有助于眼睛的生化防御系统。为了揭示眼泪中是否存在免疫介质和脂多糖结合半乳糖凝集素-3,从健康和病理状态的眼中收集眼泪样品。还开始研究正常人结膜和角膜上皮中galectin-3和galectin-3反应性糖配体的表达,以了解galectin-3在眼表病理中的作用。
方法-使用两种方法进行免疫印迹分析兔抗galectin-3多克隆抗体或小鼠单克隆抗体被用于检测泪液中galectin-3。通过使用A1D6小鼠单克隆抗体的免疫细胞化学检测组织标本中Galectin-3的表达,并使用标记的galectin-3通过凝集素组织化学观察galectin-3反应性糖配体。
结果-仅在患者的眼泪中发现了Galectin-3眼表疾病。它在正常的角膜和结膜上皮中表达,但在泪腺中不表达。在含有galectin-3的泪液中发现的炎症性白细胞和杯状细胞也表达了galectin-3。在结膜和角膜上皮细胞与desmoglein共定位的表面上检测到Galectin-3结合位点。
结论—这项研究揭示了Galectin-3在眼病患者眼泪液中的表达。可以预期这种内源性凝集素(由炎症以及上皮细胞产生)在抗菌作用和炎症调节中的作用。

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