Unknown,Transcriptomics,Genomics,Proteomics

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Expression analysis of a full Lsr locus knockout in Haemophilus influenzae strain CZ4126/02

ABSTRACT: Investigation of whole genome gene expression differences in a full (nine-gene) Lsr locus knockout strain compared to the wild-type strain (CZ4126/02). This Lsr mutant is unable to import the autoinducer-2 (AI-2) quorum sensing molecule nor mediate any potential LsrR-based transcriptional regulation. WT and Lsr-KO strains were grown in two different media to two or three different time-points. RNA was extracted from these sampling conditions in two separate cultures (biological replicates). Each of these RNA samples was run on two different H. influenzae supragenome arrays (technical replicates). 3-13 probes represent each subcluster (gene) and two replicate probe sets are included on each custom NTHi supragenome chip. Thus, under each sampling condition, 8 separate values are obtained per strain and used for WT:Lsr-KO comparisons.

ORGANISM(S): Haemophilus influenzae

SUBMITTER: Benjamin Janto

PROVIDER: E-GEOD-41690 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:Pneumonia is a serious problem worldwide. We recently demonstrated that innate defense mechanisms of the lung are highly inducible against pneumococcal pneumonia. To determine the breadth of protection conferred by stimulation of lung mucosal innate immunity, and to identify cells and signaling pathways activated by this treatment, mice were treated with an aerosolized bacterial lysate, then challenged with lethal doses of bacterial and fungal pathogens. Mice were highly protected against a broad array of Gram-positive, Gram-negative, and Class A bioterror bacterial pathogens, and Aspergillus fumigatus. Protection was associated with rapid pathogen killing within the lungs, and this effect was recapitulated in vitro using a respiratory epithelial cell line. Gene expression analysis of lung tissue showed marked activation of NF-kappaB, Type I and II interferon, and antifungal Card9-Bcl10-Malt1 pathways. Cytokines were the most strongly induced genes, but the inflammatory cytokines TNF and IL-6 were not required for protection. Lung-expressed antimicrobial peptides were also highly upregulated. Taken together, stimulated innate resistance (StIR) appears to occur through the activation of multiple host defense signaling pathways in lung epithelial cells, inducing rapid pathogen killing, and conferring broad protection against virulent bacterial and fungal pathogens. Augmentation of innate antimicrobial defenses of the lungs might have therapeutic value for protection of patients with neutropenia or impaired adaptive immunity against opportunistic pneumonia, and for defense of immunocompetent subjects against a bioterror threat or epidemic respiratory infection. Keywords: differential gene expression; time course; innate immunity; pneumonia; immunocompromised host; lung epithelium Gene expression patterns in mouse lung homogenates were analyzed 2h after exposure to aerosolized PBS (Sham treatment), 2h after exposure to aerosolized NTHi lysate or 4h after exposure to aerosolized NTHi lysate. Each group consisted of six mice.

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Expression analysis of a full Lsr locus knockout in Haemophilus influenzae strain CZ4126/02

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