Functions of Peptidoglycan Recognition Proteins (Pglyrps) at the Ocular Surface: Bacterial Keratitis in Gene-Targeted Mice Deficient in Pglyrp-2, -3 and -4

PloS One
Ranjita N GowdaShukti Chakravarti

Abstract

Functions of antimicrobial peptidoglycan recognition proteins (Pglyrp1-4) at the ocular surface are poorly understood. Earlier, we reported an antibacterial role for Pglyrp-1 in Pseudomonas aeruginosa keratitis. Here we investigated functions of three other related genes Pglyrp-2, -3 and -4 in a mouse model of P. aeruginosa keratitis. Wild type (WT) and each of the Pglyrp-null genotypes were challenged with P. aeruginosa keratitis. The eyes were scored in a blinded manner 24 and 48h post infection. Viable bacterial counts and inflammatory factors (IL-12, TNF-α, IFN-γ, CCL2, IL-6 and IL-10) were measured in whole eye homogenates using cytometric bead arrays. Expressions of Pglyrp-1-4, mouse beta defensins (mBD)-2,-3, cathelicidin-related antimicrobial peptide (CRAMP) were determined by qRTPCR in total RNA extracts of uninfected and infected eyes of WT and each of the Pglyrp-null mouse types. The Pglyrp-2-/- mice showed reduced disease and lower induction of pro-inflammatory TNF-α (p = 0.02) than WT or the other Pglyrp null mice. Viable bacterial yield was significantly lower in the Pglyrp-2-/- (p = 0.0007) and the Pglyrp-4-/- (p = 0.098) mice. With regards to expression of these antimicrobial genes, Pglyrp-2 expression was induc...Continue Reading

References

Jan 25, 2002·Nature·Michael Zasloff
Nov 26, 2003·Investigative Ophthalmology & Visual Science·Ellen J LeeSuzanne M J Fleiszig
Jun 18, 2004·Immunological Reviews·Håkan Steiner
Aug 26, 2006·Genome Biology·Roman Dziarski, Dipika Gupta
Jan 12, 2007·The Ocular Surface·Alison M McDermott
Mar 17, 2007·Nature Reviews. Microbiology·Julien Royet, Roman Dziarski
Oct 31, 2007·Immunity·Thirumala-Devi KannegantiGabriel Núñez
Apr 24, 2009·Investigative Ophthalmology & Visual Science·Amit GhoshShukti Chakravarti
Nov 15, 2011·Nature Reviews. Immunology·Julien RoyetRoman Dziarski

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Methods Mentioned

BETA
dissecting
PCR
flow cytometry

Software Mentioned

FCAP Array TM
Image J

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