ABSTRACT
The host immune responses to Staphylococcus epidermidis, a frequent cause of nosocomial infections, are not well understood. We have established a bath immersion model of this infection in zebrafish (Danio rerio) larvae. Macrophages play a primary role in the host immune response and are involved in clearance of infection in the larvae. S. epidermidis infection results in upregulation of tlr-2 There is marked inflammation characterized by heightened NF-κB signaling and elevation of several proinflammatory cytokines. There is rapid upregulation of il-1b and tnf-a transcripts, whereas an increase in il-6 levels is relatively more delayed. The IL-6 signaling pathway is further amplified by elevation of IL-6 signal transducer (il-6st) levels, which negatively correlates with miRNA dre-miR-142a-5p. Enhanced IL-6 signaling is protective to the host in this model as inhibition of the signaling pathway resulted in increased mortality upon S. epidermidis infection. Our study describes the host immune responses to S. epidermidis infection, establishes the importance of IL-6 signaling, and identifies a potential role of miR-142-5p-il-6st interaction in this infection model.
Subject(s)
Interleukin-6/metabolism , NF-kappa B/metabolism , Staphylococcal Infections/immunology , Staphylococcus epidermidis/physiology , Zebrafish Proteins/metabolism , Animals , Animals, Genetically Modified , Cells, Cultured , Cross Infection , Disease Models, Animal , Disease Resistance , Humans , Larva , MicroRNAs/genetics , NF-kappa B/genetics , Signal Transduction , Toll-Like Receptor 2/metabolism , Zebrafish , Zebrafish Proteins/geneticsABSTRACT
Ultraviolet radiation is a pervasive stimulus with wide-ranging effects on all living forms. The effects of UV vary from physiological to pathological, depending on levels of exposure, but the immune response at the organismal level is not well understood. We use the zebrafish embryo and larva to study immune responses to UV stress in vivo. UV exposure causes inflammation characterized by systemic induction of proinflammatory cytokines. Leukocytes are an important component of this systemic response and upregulate IL-1ß expression proportional to the dose of UV exposure. Increased levels of this proinflammatory cytokine counteract the lethal effect of high doses of UV.
Subject(s)
Inflammation Mediators/metabolism , Interleukin-1beta/biosynthesis , Interleukin-1beta/radiation effects , Leukocytes/radiation effects , Zebrafish/immunology , Animals , Animals, Genetically Modified , Disease Models, Animal , Embryo, Nonmammalian/immunology , Embryo, Nonmammalian/pathology , Embryo, Nonmammalian/radiation effects , Inflammation/etiology , Inflammation/genetics , Inflammation/mortality , Inflammation Mediators/radiation effects , Interleukin-1beta/genetics , Larva/genetics , Larva/immunology , Larva/radiation effects , Leukocytes/immunology , Leukocytes/pathology , NF-kappa B/genetics , NF-kappa B/metabolism , NF-kappa B/radiation effects , Signal Transduction/genetics , Signal Transduction/immunology , Signal Transduction/radiation effects , Ultraviolet Rays/adverse effects , Zebrafish/geneticsABSTRACT
MEP1A, which encodes the α subunit of meprin metalloproteinases, is a susceptibility gene for inflammatory bowel disease (IBD), and decreased intestinal meprin-α expression is associated with enhanced IBD in humans. Mice lacking meprin α (α knockout, αKO) have more severe colitis induced by dextran sulfate sodium (DSS) than wild-type (WT) mice, indicating an anti-inflammatory role for meprin A. Previous studies and those herein indicate the meprin B has proinflammatory activities. Therefore, mice lacking both meprin A and B (dKO mice) were generated to determine how their combined absence alters the inflammatory response to DSS. Unchallenged dKO mice grow and reproduce normally and have no obvious abnormal phenotype, except for a slightly elevated plasma albumin in both males and females and a lower urine creatinine level in dKO males. Upon oral administration of 3.5% DSS, the dKO mice have more severe colitis than the WT and ßKO mice but significantly less than the αKO mice. The dKO mice lose more weight and have elevated MPO and IL-6 activities in the colon compared with WT mice. Systemic inflammation, monitored by plasma nitric oxide levels, is absent in DSS-treated dKO mice, unlike WT mice. The severity of experimental IBD in dKO mice is intermediate between αKO and WT mice. The data indicate that the absence of meprin A aggravates chronic inflammation and the lack of meprin B affords some protection from injury. Manipulation of the expression of meprin gene products may have therapeutic potential.
Subject(s)
Inflammatory Bowel Diseases/metabolism , Inflammatory Bowel Diseases/pathology , Metalloendopeptidases/metabolism , Administration, Oral , Animals , Anti-Inflammatory Agents/metabolism , Chronic Disease , Colitis/metabolism , Colitis/pathology , Dextran Sulfate/administration & dosage , Disease Progression , Female , Genetic Predisposition to Disease , Inflammation Mediators/metabolism , Inflammatory Bowel Diseases/chemically induced , Inflammatory Bowel Diseases/genetics , Interleukin-6/metabolism , Intestinal Mucosa/metabolism , Male , Metalloendopeptidases/deficiency , Mice , Mice, Knockout , Permeability , Peroxidase/metabolism , Severity of Illness IndexABSTRACT
Interleukin-18 (IL-18), a pro-inflammatory cytokine, is a key factor in inflammatory bowel disease (IBD). Caspase-1 activates this cytokine, but other proteases are likely involved in maturation. Because meprin metalloproteinases have been implicated in IBD, the interaction of these proteases with proIL-18 was studied. The results demonstrate that the meprin beta subunit of meprins A and B cleaves proIL-18 into a smaller 17-kDa product. The cleavage is at the Asn51-Asp52 bond, a site C-terminal to caspase-1 cleavage. The cleavage occurred in vitro with a Km of 1.3 microm and in Madin-Darby canine kidney cells transfected with meprin beta when proIL-18 was added to the culture medium. The product of meprin B cleavage of proIL-18 activated NF-kappaB in EL-4 cells, indicating that it was biologically active. To determine the physiological significance of the interactions of meprins with proIL-18, an experimental model of IBD was produced by administering dextran sulfate sodium (DSS) to wild-type and meprin beta knock-out (betaKO) mice, and the serum levels of active IL-18 were determined. DSS-treated meprin betaKO mice had lower levels of the active cytokine in the serum compared with wild-type mice. Furthermore, in meprin alphaKO mice, which express meprin beta but not alpha, active IL-18 was elevated in the serum of DSS-treated mice compared with wild-type mice, indicating that the meprin isoforms have opposing effects on the IL-18 levels in vivo. This study identifies proIL-18 as a biologically important substrate for meprin beta and implicates meprins in the modulation of inflammation.
Subject(s)
Colitis/metabolism , Interleukin-18/metabolism , Metalloendopeptidases/metabolism , Protein Precursors/metabolism , Amino Acid Sequence , Animals , Cell Line , Colitis/chemically induced , Dextran Sulfate/pharmacology , Dogs , Humans , Interleukin-18/chemistry , Interleukin-18/genetics , Isoenzymes/metabolism , Kinetics , Male , Metalloendopeptidases/deficiency , Metalloendopeptidases/genetics , Mice , Mice, Knockout , Molecular Sequence Data , Protein Binding , Protein Precursors/genetics , RatsABSTRACT
Meprins are unique plasma membrane and secreted metalloproteinases that are highly regulated at the transcriptional and post-translational levels. Meprin alpha and beta subunits are abundantly expressed in kidney and intestinal epithelial cells, are secreted into the urinary tract and intestinal lumen, and are found in leukocytes and cancer cells under certain conditions. Their location and proteolytic activities indicate functions at the interface of the host and the external environment, and in trafficking of macrophages and metastases of cancer cells. These proteases can be detrimental when there is tissue damage or disruption, as in acute renal injury or intestinal inflammation, and there is evidence they are involved in movement of leukocytes and cancer cells to sites of infection or in metastasis, respectively.
