Bacteria induce autophagy in a human ocular surface cell line.
Bacteria induce autophagy in a human ocular surface cell line.
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DOI：
10.1016/j.exer.2017.12.010
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发表时间：
2018-03
影响因子：
3.4
通讯作者：
Shanks RMQ
中科院分区：
文献类型：
--
作者：
Brothers KM;Kowalski RP;Tian S;Kinchington PR;Shanks RMQ
自噬保护细胞免受细胞内病原体的侵害，但也可能被一些感染因子利用。目前尚不清楚细菌是否会诱导角膜细胞的自体增生。本研究的目的是建立一种眼表自噬报告细胞系，并确定眼部细菌病原体是否通过自噬诱导影响宿主反应。使用人角膜缘上皮（HCLE）细胞中LC 3-GFP融合蛋白的慢病毒转导制备细胞系。自噬抑制剂3-甲基腺嘌呤（3-methyladenine，3′MA）和巴弗洛霉素（bafilomycin）可抑制雷帕霉素和氯化铵诱导的LC 3-GFP斑点。重要的是，发现来自大肠杆菌、粘质沙雷氏菌、金黄色葡萄球菌、甲氧西林敏感（MSSA）和耐甲氧西林（MRSA）的分泌体诱导自噬，而其他细菌，包括鲍曼不动杆菌、木糖氧化无色杆菌、粪肠球菌、肺炎克雷伯氏菌、莫拉氏菌属、而嗜麦芽窄食单胞菌则没有。我们的数据表明测试的MRSA和MSSA的眼部分离株在自噬激活方面的差异。经3′MA处理的HCLE对S.粘质杆菌和MRSA角膜炎分离株，相比之下，巴弗洛霉素A1治疗没有引起差异。这项工作证明了自噬报告角膜细胞系的成功开发和验证，并表明眼部细菌分离株在自噬激活方面的差异。
Autophagy protects cells from intracellular pathogens, but can be exploited by some infectious agents to their benefit. Currently it is not known if bacteria induce autohpagy in cells of the cornea. The goal of this study was to develop an ocular surface autophagy reporter cell line and determine whether ocular bacterial pathogens influence host responses through autophagy induction. The cell line was made using lentivirus transduction of an LC3-GFP fusion protein in human corneal limbal epithelial (HCLE) cells. LC3-GFP puncta in HCLEs were induced by rapamycin and ammonium chloride treatments, and prevented by the autophagy inhibitors 3-methyladenine (3′MA) and bafilomycin. Importantly, secretomes from Escherichia coli, Serratia marcescens, Staphylococcus aureus, methicillin sensitive (MSSA) and resistant (MRSA), were found to induce autophagy, whereas other bacteria, including Acinetobacter baumannii, Achromobacter xylosoxidans, Enterococcus faecalis, Klebsiella pneumoniae, Moraxella sp., and Stenotrophomonas maltophilia, did not. Our data indicates differences between tested ocular isolates of MRSA and MSSA in the activation of autophagy. HCLEs treated with 3′MA were slightly more susceptible to cytotoxic factors produced by S. marcescens and MRSA keratitis isolates, by contrast, bafilomycin A1 treatment caused no difference. This work demonstrates the successful development and validation of an autophagy reporter corneal cell line and indicates differences between ocular bacterial isolates in the activation of autophagy.
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影响因子：
3.4
作者：
Kowalski RP;Romanowski EG;Mah FS;Shanks RM;Gordon YJ
通讯作者：
Gordon YJ
影响因子：
3.8
作者：
Lakhundi, Sahreena;Siddiqui, Ruqaiyyah;Khan, Naveed Ahmed
通讯作者：
Khan, Naveed Ahmed
DOI：
10.1016/0304-4165(76)90146-x
发表时间：
1976-01-01
期刊：
BIOCHIMICA ET BIOPHYSICA ACTA
影响因子：
--
作者：
AMENTA, JS;BACCINO, FM;SARGUS, MJ
通讯作者：
SARGUS, MJ
DOI：
10.1136/bjophthalmol-2016-308487
发表时间：
2017-03
期刊：
The British journal of ophthalmology
影响因子：
--
作者：
Lalitha P;Manoharan G;Karpagam R;Prajna NV;Srinivasan M;Mascarenhas J;Das M;Porco TC;Lietman TM;Cevallos V;Keenan JD
通讯作者：
Keenan JD
影响因子：
5.4
作者：
Leib, David A.;Alexander, Diane E.;Ferguson, Thomas A.
通讯作者：
Ferguson, Thomas A.