Methods for In Vivo/Ex Vivo Analysis of Antimicrobial Peptides in Bacterial Keratitis: siRNA Knockdown, Colony Counts, Myeloperoxidase, Immunostaining, and RT-PCR Assays

Satya Sree Kolar, Hasna Baidouri, Maria Luisa Mangoni, Alison McDermott

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

Abstract

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.
Original languageEnglish
Title of host publicationAntimicrobial Peptides
EditorsPaul Hansen
Place of PublicationLondon
PublisherSpringer
Pages411-425
Volume1548
ISBN (Print)978-1-4939-6735-3
DOIs
Publication statusPublished - 25 Dec 2016

Publication series

NameMethods in Molecular Biology
PublisherSpringer
ISSN (Electronic)1064-3745

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