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

Kolar, Satya Sree, Baidouri, Hasna, Mangoni, Maria Luisa and McDermott, Alison (2016) Methods for In Vivo/Ex Vivo Analysis of Antimicrobial Peptides in Bacterial Keratitis: siRNA Knockdown, Colony Counts, Myeloperoxidase, Immunostaining, and RT-PCR Assays. In: Antimicrobial Peptides. Methods in Molecular Biology, 1548 . Springer, London, pp. 411-425. ISBN 978-1-4939-6735-3

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Official URL: https://doi.org/10.1007/978-1-4939-6737-7_30

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.

Item Type: Book Section
Uncontrolled Keywords: Antimicrobial peptide, Inflammation, Keratitis, Neutrophil, Pseudomonas
Subjects: B500 Ophthalmics
Department: Faculties > Health and Life Sciences > School of Life Sciences > Applied Sciences
Depositing User: Becky Skoyles
Date Deposited: 19 May 2017 14:39
Last Modified: 19 May 2017 14:39
URI: http://nrl.northumbria.ac.uk/id/eprint/30773

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