Unknown,Transcriptomics,Genomics,Proteomics

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Transcription profiling of wild type Listera monocytogenes EGD-e and an isogenic deltahlydeltaplcA mutant strain 1 hour or 4 hours post infection into human intestinal epithelial cells

ABSTRACT: We infected wild type L. monocytogenes EGD-e (1) and its isogenic deltahlydeltaplcA (2) (lacking the ability to breach the vacuolar compartment of host cells following uptake) mutant strain to human intestinal epithelial cell line (Caco-2) with an MOI of 100 and 500 respectively. Bacterial total RNA was isolated at 1 h (deltahlydeltaplcA) and 4 h (EGD-e) post infection, reverse transcribed, hybridised to whole genome microarray and microarray data was analysed as described previously (3) 1. Glaser et al. 2001. Comparative genomics of Listeria species. Science 294:849-852. 2. Paschen et al. 2000. Human dendritic cells infected by Listeria monocytogenes: induction of maturation, requirements for phagolysosomal escape and antigen presentation capacity. Eur.J.Immunol. 30:3447-3456. 3. Chatterjee et al. 2006. Intracellular gene expression profile of Listeria monocytogenes. Infect.Immun. 74:1323-1338.

INSTRUMENT(S): Generation III Array Scanner [Amersham]

ORGANISM(S): Listeria monocytogenes

SUBMITTER: Torsten Hain

PROVIDER: E-MEXP-1144 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:Objective: Reg3g has been proposed to have a protective role against infection due to its bactericidal effect on Gram-positive bacteria, but evidence from in vivo studies is lacking. Therefore we generated a Reg3g-/- mouse, to determine its role in intestinal homeostasis and protection against experimental infection. Methods: Reg3g-/- mice were phenotyped using histological methods and a range of innate and immune markers. To investigate the antimicrobial role of Reg3g we experimentally infected mice with Gram-positive Listeria monocytogenes and Gram-negative Salmonella entertitidis and measured translocated bacteria, mucosal and systemic markers of infection. Results: Reg3g-/- mice display altered ileal mucus distribution and increased bacterial contact with the epithelium. , concomitant with This increased the inflammatory status in of the ileal mucosa and increased expression of Il-22, myeloperoxidase (MPO) and serum chemokines in serum. In response to infection, Reg3g-/- mice showed transcriptome changes and elevated levels of mucosal MPO in the ileum, but no increased bacterial translocation to the organs. Conclusions: Reg3g is equally distributed throughout the mucus of wild type (wt) mice and its absence results in an altered distribution of the ileal mucus. Reg3g deficiency also results resulted in increased bacterial contact with the epithelium and heightened inflammatory responses in the ileal mucosa. We propose that Reg3g binds pathogens suggesting it and contributes to mucus barrier function by ensnaring bacteria. Compared to wt mice, Reg3g-/- mice infected with S. enteritidis and L. monocytogenes show an increase of mucosal inflammatory markers indicating the protective, anti-microbial roles of Reg3g in defence against both Gram-positive and -negative bacteria. This study was set up according to a one-treatment, one-control design; treatments were inoculation with either Listeria monocytogenes or Salmonella enteritidis bacterial pathogens. The study results contain transcriptional profiles from infected and sham-infected control C57Bl/6 mice. In total, this study includes data from 2 treatments and 1 control of (pooled) wild-type C57Bl/6 mice and Reg3g-/- KO mutant C57Bl/6 mice = 6 arrays.

Dataset Information

Transcription profiling of wild type Listera monocytogenes EGD-e and an isogenic deltahlydeltaplcA mutant strain 1 hour or 4 hours post infection into human intestinal epithelial cells

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