Generation of Mouse Small Intestinal Epithelial Cell Lines That Allow the Analysis of Specific Innate Immune Functions

Schwerk, Johannes; Köster, Mario; Hauser, Hansjörg; Rohde, Manfred; Fulde, Marcus; Hornef, Mathias W.; May, Tobias

doi:10.1371/journal.pone.0072700

Cited by 26 publications

(38 citation statements)

References 46 publications

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“…The mouse small intestine cell line (ICE-1) was a kind gift from InScreenex (InSCREENeX, Braunschweig, Germany) (Schwerk et al, 2013). Cells were cultured in a defined medium supplied by the manufacturer.…”

Section: Methodsmentioning

confidence: 99%

The Novel Type 1 Fimbriae FimH Receptor Calreticulin Plays a Role in Salmonella Host Specificity

Grzymajło

Ugorski

Suchański

et al. 2017

Front. Cell. Infect. Microbiol.

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It was suggested that minor differences in the structure of FimH are most likely associated with differences in its adhesion specificities and may determine the tropism of various Salmonella serovars to different species and tissues. We have recently shown that FimH adhesins from host-adapted serovars, e.g., Salmonella Choleraesuis (SCh), bind to other glycoprotein receptors compared to FimH from host-unrestricted Salmonella Enteritidis (SE). Here we identify porcine calreticulin expressed by swine intestinal cells as a host-specific receptor for SCh FimH adhesin, suggesting that such an interaction may contribute to SCh host specificity. Calreticulin was identified by 2D electrophoresis and mass spectrometry as a glycoprotein that was bound specifically by recombinant SCh FimH protein, but not by FimH from SE. The functionality of calreticulin as a specific receptor of SCh FimH adhesin was further confirmed by adhesion and invasion of mutated strains of SCh carrying different variants of FimH proteins to IPEC-J2 cells with overexpression and silenced expression of calreticulin. It was found that SCh carrying the active variant of FimH adhered and invaded IPEC-J2 cells with calreticulin overexpression at significantly higher numbers than those of SCh expressing the non-active variant or SE variant of FimH. Moreover, binding of SCh carrying the active variant of FimH to IPEC-J2 with silenced calreticulin expression was significantly weaker. Furthermore, we observed that SCh infection induces translocation of calreticulin to cell membrane. All of the aforementioned results lead to the general conclusion that Salmonella host specificity requires not only special mechanisms and proteins expressed by the pathogen but also specifically recognized receptors expressed by a specific host.

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Section: Methodsmentioning

confidence: 99%

The Novel Type 1 Fimbriae FimH Receptor Calreticulin Plays a Role in Salmonella Host Specificity

Grzymajło

Ugorski

Suchański

et al. 2017

Front. Cell. Infect. Microbiol.

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“…The mouse small intestine cell line IEC-1 (35) or the human Lenti-X 293T cell line was seeded into 6-well plates at a density of 2 3 10 5 cells per well in DMEM containing 10% fetal calf serum, with 1% L-glutamine and 50 mM 2-ME, in the case of IEC-1. The next day, the cells were transfected with either 25 nM of each murine small interfering RNA (siRNA) (Per1: s71484, Per2: s71485), 25 nM of each human siRNA (Per1: s10302, Per2: s16930), or 50 nM of negative control siRNA (4390844), for 48 h with the Lipofectamine 2000 Transfection Reagent (Thermo Fisher Scientific, Darmstadt, Germany) according to the manufacturer's instructions.…”

Section: Small Interfering Rna Transfectionmentioning

confidence: 99%

Circadian rhythm disruption impairs tissue homeostasis and exacerbates chronic inflammation in the intestine

et al. 2017

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Endogenous circadian clocks regulate 24-h rhythms of physiology and behavior. Circadian rhythm disruption (CRD) is suggested as a risk factor for inflammatory bowel disease. However, the underlying molecular mechanisms remain unknown. Intestinal biopsies from Per1/2 mutant and wild-type (WT) mice were investigated by electron microscopy, immunohistochemistry, and bromodeoxyuridine pulse-chase experiments. TNF-α was injected intraperitoneally, with or without necrostatin-1, into Per1/2 mice or rhythmic and externally desynchronized WT mice to study intestinal epithelial cell death. Experimental chronic colitis was induced by oral administration of dextran sodium sulfate. , caspase activity was assayed in Per1/2-specific small interfering RNA-transfected cells. was overexpressed to study antiapoptosis and the cell cycle. Genetic ablation of circadian clock function or environmental CRD in mice increased susceptibility to severe intestinal inflammation and epithelial dysregulation, accompanied by excessive necroptotic cell death and a reduced number of secretory epithelial cells. Receptor-interacting serine/threonine-protein kinase (RIP)-3-mediated intestinal necroptosis was linked to increased mitotic cell cycle arrest Per1/2-controlled Wee1, resulting in increased antiapoptosis cellular inhibitor of apoptosis-2. Together, our data suggest that circadian rhythm stability is pivotal for the maintenance of mucosal barrier function. CRD increases intestinal necroptosis, thus rendering the gut epithelium more susceptible to inflammatory processes.-Pagel, R., Bär, F., Schröder, T., Sünderhauf, A., Künstner, A., Ibrahim, S. M., Autenrieth, S. E., Kalies, K., König, P., Tsang, A. H., Bettenworth, D., Divanovic, S., Lehnert, H., Fellermann, K., Oster, H., Derer, S., Sina, C. Circadian rhythm disruption impairs tissue homeostasis and exacerbates chronic inflammation in the intestine.

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“…In the absence of crypts, epithelial proliferation and renewal are markedly diminished in the neonate animal. Cell lines and primary epithelial cells isolated from fetal intestinal tissue express innate immune receptors, respond to microbial ligands, and secrete pro-inflammatory chemoattractants ( 14 – 17 ). Regulatory mechanisms must, therefore, exist to prevent inappropriate immune stimulation.…”

Section: Innate Immune Signaling During Ontogeny: Age-dependent Recepmentioning

confidence: 99%

Ontogeny of Intestinal Epithelial Innate Immune Responses

Hornef

Fulde

2014

Front. Immunol.

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Emerging evidence indicates that processes during postnatal development might significantly influence the establishment of mucosal host-microbial homeostasis. Developmental and adaptive immunological processes but also environmental and microbial exposure early after birth might thus affect disease susceptibility and health during adult life. The present review aims at summarizing the current understanding of the intestinal epithelial innate immune system and its developmental and adaptive changes after birth.

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Generation of Mouse Small Intestinal Epithelial Cell Lines That Allow the Analysis of Specific Innate Immune Functions

Cited by 26 publications

References 46 publications

The Novel Type 1 Fimbriae FimH Receptor Calreticulin Plays a Role in Salmonella Host Specificity

The Novel Type 1 Fimbriae FimH Receptor Calreticulin Plays a Role in Salmonella Host Specificity

Circadian rhythm disruption impairs tissue homeostasis and exacerbates chronic inflammation in the intestine

Ontogeny of Intestinal Epithelial Innate Immune Responses

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