Kolar Satya Sree, Baidouri Hasna, Mangoni Maria Luisa, McDermott Alison M
The Ocular Surface Institute, College of Optometry, University of Houston, 4901 Calhoun Road, Houston, TX, 77204, USA.
Department of Biochemical Sciences "A. Rossi Fanelli", Istituto Pasteur-Fondaziona Cenci Bolognetti, Sapienza University of Rome, Rome, Italy.
Methods Mol Biol. 2017;1548:411-425. doi: 10.1007/978-1-4939-6737-7_30.
Antimicrobial peptides (AMPs) are essential components of the innate immune response. They have direct killing ability as well as immunomodulatory functions. Here, we describe techniques to identify specific AMPs involved in the protection against microbial keratitis, a vision threatening infection of the cornea of the eye which is the most serious complication of contact lens wear. Specifically we detail the use of siRNA technology to temporarily knockdown AMP expression at the murine ocular surface in vivo and then describe ex vivo assays to determine the level of bacteria, relative number of neutrophils, and levels of cytokines, chemokines, and AMPs in infected corneas.
抗菌肽（AMPs）是先天免疫反应的重要组成部分。它们具有直接杀伤能力以及免疫调节功能。在此，我们描述了鉴定参与预防微生物性角膜炎的特定抗菌肽的技术，微生物性角膜炎是一种威胁视力的眼部角膜感染，是隐形眼镜佩戴最严重的并发症。具体而言，我们详细介绍了利用小干扰RNA（siRNA）技术在体内暂时敲低小鼠眼表抗菌肽的表达，然后描述了体外试验，以确定感染角膜中的细菌水平、中性粒细胞相对数量以及细胞因子、趋化因子和抗菌肽的水平。
