Use of genome-wide expression profiling and mutagenesis to study the intestinal lifestyle of Campylobacter jejuni. Infect Immun

Department of Veterinary Pathobiology, College of Veterinary Medicine, Oklahoma State University, Stillwater, OK 74078, USA.
Infection and Immunity (Impact Factor: 4.16). 04/2005; 73(3):1797-810. DOI: 10.1128/IAI.73.3.1797-1810.2005
Source: PubMed

ABSTRACT Campylobacter jejuni is the most common bacterial cause of diarrhea worldwide. To colonize the gut and cause infection, C. jejuni must successfully compete with endogenous microbes for nutrients, resist host defenses, persist in the intestine, and ultimately infect the host. These challenges require the expression of a battery of colonization and virulence determinants. In this study, the intestinal lifestyle of C. jejuni was studied using whole-genome microarray, mutagenesis, and a rabbit ileal loop model. Genes associated with a wide range of metabolic, morphological, and pathological processes were expressed in vivo. The in vivo transcriptome of C. jejuni reflected its oxygen-limited, nutrient-poor, and hyperosmotic environment. Strikingly, the expression of several C. jejuni genes was found to be highly variable between individual rabbits. In particular, differential gene expression suggested that C. jejuni extensively remodels its envelope in vivo by differentially expressing its membrane proteins and by modifying its peptidoglycan and glycosylation composition. Furthermore, mutational analysis of seven genes, hspR, hrcA, spoT, Cj0571, Cj0178, Cj0341, and fliD, revealed an important role for the stringent and heat shock response in gut colonization. Overall, this study provides new insights on the mechanisms of gut colonization, as well as possible strategies employed by Campylobacter to resist or evade the host immune responses.

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    • "The outer membrane protein Cj0178 (CtuA) was characterized as a receptor for the iron-binding host glycoproteins ferri-transferrin, ferri-lactoferrin and ferri-ovotransferrin (Miller et al., 2008). A cj0178 mutant strain of C. jejuni NCTC 11168 showed a severe colonization defect in chicken infection experiments (Palyada et al., 2004) and a slightly attenuated phenotype in the rabbit ileal loop model (Stintzi et al., 2005). The chuABCDZ (cj1613c-cj1617) gene cluster is widespread in C. jejuni and encodes for an iron uptake system that facilitates the utilization of the host compounds like hemoglobin and hemin as iron sources (Pickett et al., 1992; Ridley et al., 2006). "
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    ABSTRACT: During the last decade Campylobacter jejuni has been recognized as the leading cause of bacterial gastroenteritis worldwide. This facultative intracellular pathogen is a member of the Epsilonproteobacteria and requires microaerobic atmosphere and nutrient rich media for efficient proliferation in vitro. Its catabolic capacity is highly restricted in contrast to Salmonella Typhimurium and other enteropathogenic bacteria because several common pathways for carbohydrate utilization are either missing or incomplete. Despite these metabolic limitations, C. jejuni efficiently colonizes various animal hosts as a commensal intestinal inhabitant. Moreover, C. jejuni is tremendously successful in competing with the human intestinal microbiota; an infectious dose of few hundreds bacteria is sufficient to overcome the colonization resistance of humans and can lead to campylobacteriosis. Besides the importance and clear clinical manifestation of this disease, the pathogenesis mechanisms of C. jejuni infections are still poorly understood. In recent years comparative genome sequence, transcriptome and metabolome analyses as well as mutagenesis studies combined with animal infection models have provided a new understanding of how the specific metabolic capacity of C. jejuni drives its persistence in the intestinal habitat of various hosts. Furthermore, new insights into the metabolic requirements that support the intracellular survival of C. jejuni were obtained. Because C. jejuni harbors distinct properties in establishing an infection in comparison to pathogenic Enterobacteriaceae, it represents an excellent organism for elucidating new aspects of the dynamic interaction and metabolic cross talk between a bacterial pathogen, the microbiota and the host.
    Frontiers in Cellular and Infection Microbiology 09/2014; 4. DOI:10.3389/fcimb.2014.00137 · 2.62 Impact Factor
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    • "Using omics data, information on cellular physiology can be obtained for microbes while they are present in the host. Such data provide crucial information on survival likelihood of pathogenic microbes and their expression of specific virulence genes (Resnik et al., 2006; Stinzi et al., 2005) as well as on beneficial effects of probiotic bacteria (van Baarlen et al., 2011). This information, combined with the host immune response, should facilitate the determination of the doseeresponse relationship in health and disease. "
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    • "To note, analysis and validation of gene expression by RT-PCR was not possible due to the low amount of isolated RNA, a problem especially challenging for in vivo experiments. Nevertheless, the same microarray platform and protocols were previously shown to produce data with high level of concordance with quantitative RT-PCR (Stintzi, 2003; Palyada et al., 2004; Stintzi et al., 2005; Reid et al., 2008a,b; Palyada et al., 2009). Overall, it was found that 449 genes were differentially expressed in vivo as compared to in vitro conditions in at least one of the experimental time points (2, 6, 24, or 48 h) using the RTC model (Figures 1 and 2). "
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