Ca2+ release from host intracellular stores and related signal transduction during Campylobacter jejuni 81–176 internalization into human intestinal cell

Laboratory of Enteric and Sexually Transmitted Diseases, FDA-Center for Biologics Evaluation and Research, 29 Lincoln Drive, Bldg 29/420 HFM440, Bethesda, MD 20892, USA.
Microbiology (Impact Factor: 2.84). 10/2005; 151(Pt 9):3097-105. DOI: 10.1099/mic.0.27866-0
Source: PubMed

ABSTRACT Campylobacter jejuni is the leading bacterial cause of human diarrhoeal disease in many parts of the world, including the USA. The ability of C. jejuni to invade the host intestinal epithelium is an important determinant of virulence. A common theme among pathogenic invasive micro-organisms is their ability to usurp the eukaryotic cell-signalling systems both to allow for invasion and to trigger disease pathogenesis. Ca(2+) is very important in a great variety of eukaryotic cell-signalling processes (e.g. calmodulin-activated enzymes, nuclear transcriptional upregulation, and cytoskeletal rearrangements). This study analyses the effects of Ca(2+) availability on invasion of human INT407 intestinal epithelial cells by C. jejuni strain 81-176. The ability of C. jejuni to invade INT407 cells was not blocked by chelation of any remaining extracellular Ca(2+) from host cells incubated in Ca(2+)-free, serum-free media. In contrast, C. jejuni invasion was markedly reduced either by chelating host intracellular Ca(2+) with 1,2-bis-(2-)ethane-N,N,N',N'-tetraacetic acid acetoxymethyl ester (BAPTA, AM) or by blocking the release of Ca(2+) from intracellular stores with dantrolene or U73122. Moreover, Bay K8644, a plasma-membrane Ca(2+)-channel agonist, was observed to stimulate C. jejuni invasion, presumably by increasing host intracellular free Ca(2+) levels. Measurement of host-cell cytosolic Ca(2+) via spectrofluorimetry and fluorescence microscopy revealed an increase in Ca(2+) from 10 min post-infection. Monolayer pretreatment with either a calmodulin antagonist or a specific inhibitor of protein kinase C was found to cause a marked reduction in C. jejuni invasion, suggesting roles for these Ca(2+)-activated modulators in signal-transduction events involved in C. jejuni invasion. These results demonstrate that C. jejuni induces the mobilization of Ca(2+) from host intracellular stores, which is an essential step in the invasion of intestinal cells by this pathogen.

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    • "For instance, intracellular free calcium ion (Ca2+), an important intracellular messenger with multiple physiological functions, is increased when cells are infected with some bacterial pathogens [20]. Thus, Helicobacter pylori or Campylobacter jejuni cause an elevation of intracellular free Ca2+ concentration ([Ca2+]i) through Ca2+ release and/or influx mechanisms in gastric mucous epithelial cells or in intestinal epithelial cells during infection [21], [22]. The high [Ca2+]i in macrophages caused by infection with Listeria monocytogenes or Brucella abortus are involved in bacterial invasion and escape from phagocytotic vesicles for intracellular replication [23], [24]. "
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    ABSTRACT: Leptospira-induced macrophage death has been confirmed to play a crucial role in pathogenesis of leptospirosis, a worldwide zoonotic infectious disease. Intracellular free Ca(2+) concentration ([Ca(2+)]i) elevation induced by infection can cause cell death, but [Ca(2+)]i changes and high [Ca(2+)]i-induced death of macrophages due to infection of Leptospira have not been previously reported. We first used a Ca(2+)-specific fluorescence probe to confirm that the infection of L. interrogans strain Lai triggered a significant increase of [Ca(2+)]i in mouse J774A.1 or human THP-1 macrophages. Laser confocal microscopic examination showed that the [Ca(2+)]i elevation was caused by both extracellular Ca(2+) influx through the purinergic receptor, P2X7, and Ca(2+) release from the endoplasmic reticulum, as seen by suppression of [Ca(2+)]i elevation when receptor-gated calcium channels were blocked or P2X7 was depleted. The LB361 gene product of the spirochete exhibited phosphatidylinositol phospholipase C (L-PI-PLC) activity to hydrolyze phosphatidylinositol-4,5-bisphosphate (PIP2) into inositol-1,4,5-trisphosphate (IP3), which in turn induces intracellular Ca(2+) release from endoplasmic reticulum, with the Km of 199 µM and Kcat of 8.566E-5 S(-1). Secretion of L-PI-PLC from the spirochete into supernatants of leptospire-macrophage co-cultures and cytosol of infected macrophages was also observed by Western Blot assay. Lower [Ca(2+)]i elevation was induced by infection with a LB361-deficient leptospiral mutant, whereas transfection of the LB361 gene caused a mild increase in [Ca(2+)]i. Moreover, PI-PLCs (PI-PLC-β3 and PI-PLC-γ1) of the two macrophages were activated by phosphorylation during infection. Flow cytometric detection demonstrated that high [Ca(2+)]i increases induced apoptosis and necrosis of macrophages, while mild [Ca(2+)]i elevation only caused apoptosis. This study demonstrated that L. interrogans infection induced [Ca(2+)]i elevation through extracellular Ca(2+) influx and intracellular Ca(2+) release cause macrophage apoptosis and necrosis, and the LB361 gene product was shown to be a novel PI-PLC of L. interrogans responsible for the [Ca(2+)]i elevation.
    PLoS ONE 10/2013; 8(10):e75652. DOI:10.1371/journal.pone.0075652 · 3.23 Impact Factor
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    • "). Invasion of human intestinal epithelial cells by C. jejuni in vitro requires release of Ca 2+ from intracellular stores and inhibition of calmodulin using a calmodulin agonist inhibits C. jejuni intracellular epithelial cell invasion in vitro (Hu et al. 2005). This finding suggests there may be a relationship with the association observed surrounding the T-cadherin gene, although no correlation between SNP genotypes from the two regions was found. "
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    ABSTRACT: The enteropathogen Campylobacter jejuni is a major worldwide health and economic burden, being one of the leading causes of bacterial gastroenteritis and commonly linked to postinfectious onset of autoimmune disease. Chickens are a major vector for human infection and even though variation in avian colonization level is heritable, no previous studies have identified regions of the genome associated with colonization resistance. We performed a genome-wide association study of resistance to C. jejuni colonization in the avian intestine, controlling for population structure which revealed a risk locus with genome-wide significance spanning the T-cadherin (CDH13) gene. A second possible risk locus was also identified close to calmodulin (CALM1), a calcium-activated modulator of cadherin function. In addition, gene expression analysis of mRNA sequencing profiles revealed that the relative expression of the two genes is significantly associated with colonization resistance. Functional studies have previously demonstrated involvement of cadherins and calmodulin in C. jejuni intracellular invasion and colonization of human intestinal epithelial cells in vitro. Consistent with this, our analysis reveals that variation surrounding these genes is associated with avian colonization resistance in vivo and highlights their potential as possible targets for control of the bacterium in avian and human populations.
    G3-Genes Genomes Genetics 03/2013; 3(5). DOI:10.1534/g3.113.006031 · 2.51 Impact Factor
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    • "For example, invasion of Shigella flexneri in epithelial cells induces an increase in intracellular Ca 2þ , but cytoskeletal rearrangements are also observed in cells with no detectable Ca 2þ response [20]. Listeria monocytogenes and Campylobacter jejuni induce intracellular Ca 2þ changes in host cells [1] [21], but while chelation of intracellular Ca 2þ interferes with C. jejuni internalization [1], it has no effect on L. monocytogenes invasion [21]. Contradictory results regarding the dependence on cytosolic Ca 2þ changes for intracellular invasion by Salmonella typhimurium and stimulation of IL-8 production have been also reported [2] [22]. "
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    ABSTRACT: Calcium signaling has been implicated in various steps in bacterial pathogenesis. Here, we investigated the role of Ca(2+) signaling in intracellular invasion of non-phagocytic host cells infected with Orientia tsutsugamushi, the causative agent of scrub typhus. The bacteria induced a transient Ca(2+) increase in HeLa cells immediately after infection and the source of the mobilized Ca(2+) appears to be intracellular stores, not the extracellular milieu, as determined using extracellular (EGTA) or intracellular (BAPTA-AM) Ca(2+) chelators. O. tsutsugamushi rapidly induced activation of PLC-γ1, as indicated by tyrosine phosphorylation. PLC-γ1 activity increased within 1 min of infection and returned to the basal level by 5 min. Pretreatment of host cells with inhibitors of PLC-γ1 (U73122) or IP3R channel activity (2-APB) significantly blocked bacterial entry without affecting bacterial attachment. In addition, these chemical inhibitors were effective in suppressing not only the activation of ERK MAP kinase but also the expression of the chemokine MCP-1. Taken together, Ca(2+) signaling induced by O. tsutsugamushi may play a crucial role in bacterial pathogenesis including inflammatory reactions as well as intracellular invasion into non-phagocytic host cells.
    Microbial Pathogenesis 02/2011; 50(6):326-30. DOI:10.1016/j.micpath.2011.02.007 · 2.00 Impact Factor
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