Carbohydrate Determinants in Ferret Conjunctiva are Affected by Infection with Influenza H1N1 Virus
Carbohydrate Determinants in Ferret Conjunctiva are Affected by Infection with Influenza H1N1 Virus
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DOI：
10.3109/02713683.2013.803289
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发表时间：
2013-10-01
影响因子：
2
通讯作者：
Vorum, Henrik
中科院分区：
文献类型：
--
作者：
Kirkeby, Svend;Martel, Cyril J. M.;Vorum, Henrik
背景：碳水化合物通常作为微生物的细胞表面受体，流感病毒优先通过病毒血凝素与唾液酸结合。该病毒不仅可以附着在呼吸道上皮上，还可以附着在眼表上皮上。目的：确定雪貂是否可以用于研究病毒引起的结膜炎，并评估流感发作期间结膜糖基化模式的变化。方法：通过鼻腔接种使雪貂感染H1N1流感病毒。采用凝集素和免疫组织化学方法检测感染后0至10天雪貂眼睑切片中原位碳水化合物的表达。结果：接种后5-6天，结膜细胞肥大，出现PAS阳性细胞和PAS+阿尔新蓝染色细胞。研究了三种唾液酸检测凝集素的结合：WGA、MAA2 和 SNA1。虽然它们都没有对未感染雪貂的结膜上皮细胞进行任何程度的染色，但在与所有三种凝集素一起孵育后，感染雪貂中出现阳性结膜反应。 MUC1 抗体的结合似乎可以检测粘蛋白分子中的唾液酸化决定簇，表明 MUC1 在流感感染开始时在大多数鳞状结膜细胞中从头表达。 MUC5AC阳性上皮细胞，可能是杯状细胞，在患病的结膜中增殖。结论：雪貂经鼻接种H1N1病毒对结膜细胞有影响，改变其聚糖的表达。合成聚糖是泪膜的组成部分，本研究有助于揭示眼睑表面上皮发生的变化，从而阐明病毒介导的结膜炎的病理生理学。雪貂是研究人类流感病毒介导的人类结膜炎的合适动物模型。
Background: Carbohydrates often accomplish as cell-surface receptors for microorganisms and influenza virus preferentially binds to sialic acid through the viral haemagglutinin. The virus may attach not only to the epithelium in the airways, but also to the surface ocular epithelium.Purpose: To decide if ferrets can be used to study virus induced conjunctivitis and to evaluate changes in the conjunctival glycosylation pattern during an influenza attack.Methods: Ferrets were infected with H1N1 influenza virus via nasal inoculation. The in situ carbohydrate expressions in eyelid sections from ferrets 0 to 10 days after infection was examined using lectin- and immunohistochemistry.Results: The conjunctival cells became hypertrophic with appearance of both PAS positive and PAS+Alcian Blue stained cells 5-6 days after inoculation. The binding of three sialic acid detecting lectins were investigated: WGA, MAA2 and SNA1. While none of them stained conjunctival epithelial cells in the non-infected ferrets to any extent, there was a positive conjunctival reaction in the infected ferret after incubation with all three lectins. Binding of a MUC1 antibody that seems to detect sialylated determinants in the mucin molecule indicates that MUC1 is de novo expressed in most of the squamous conjunctival cells at the start of the influenza infection. MUC5AC positive epithelial cells, probably goblet cells, proliferate in the diseased conjunctiva.Conclusion: Nasal inoculation of H1N1 virus to ferrets has an effect on the conjunctival cells and change their expression of glycans. Synthesized glycans are an integral part of the tear film and the present study contributes to reveal the changes that occur in the surface epithelium in the eyelid and thereby to elucidate the pathophysiology of the virus mediated conjunctivitis. Ferrets are suitable animal models to study human conjunctivitis mediated by human influenza virus.