Differential regulation of neutrophil-activating chemokines by IL-6 and its soluble receptor isoforms.

Interleukin-6 signaling via its soluble receptor (sIL-6R) differentially regulates inflammatory chemokine expression and leukocyte apoptosis to coordinate transition from neutrophil to mononuclear cell infiltration. sIL-6R activities may, however, be influenced in vivo by the occurrence of two sIL-6R isoforms that are released as a consequence of differential mRNA splicing (DS) or proteolytic cleavage (PC) of the cognate IL-6R (termed DS- and PC-sIL-6R). Using human peritoneal mesothelial cells and a murine model of peritoneal inflammation, studies described in this work have compared the ability of both isoforms to regulate neutrophil recruitment. In this respect, DS- and PC-sIL-6R were comparable in their activities; however, these studies emphasized that IL-6 trans signaling differentially controls neutrophil-activating CXC chemokine expression. In vitro, stimulation of mesothelial cells with IL-6 in combination with either DS-sIL-6R or PC-sIL-6R showed no induction of CXC chemokine ligand (CXCL)1 (GRO alpha) and CXCL8 (IL-8), whereas both isoforms enhanced CXCL5 (ENA-78) and CXCL6 (granulocyte chemotactic protein-2) expression. Moreover, when complexed with IL-6, both isoforms specifically inhibited the IL-1 beta-induced secretion of CXCL8. These findings were paralleled in vivo, in which induction of peritoneal inflammation in IL-6-deficient (IL-6(-/-)) mice resulted in enhanced keratinocyte-derived chemokine and macrophage-inflammatory protein-2 (the murine equivalent of CXCL1 and CXCL8) levels, but reduced LPS-induced CXC chemokine (the murine equivalent of CXCL5) expression. Reconstitution of IL-6 signaling in IL-6(-/-) mice with IL-6 and its soluble receptor isoforms corrected this chemokine imbalance and suppressed overall neutrophil infiltration. These data confirm that sIL-6R-mediated signaling primarily limits neutrophil influx; however, induction of CXCL5 and CXCL6 may regulate other neutrophil responses.

Glucose degradation products (GDP) retard remesothelialization independently of D-glucose concentration.

BACKGROUND: Glucose degradation products (GDP) present in heat-sterilized dialysis fluids are thought to contribute to cellular dysfunction and membrane damage during peritoneal dialysis. To examine the effects of specific GDP on the remesothelialization process, the impact of conventional and low GDP peritoneal dialysis solutions, D-glucose, and individual GDP in a scratch-wounding model was assessed. METHODS: Scratch (0.5 to 0.6 mm)-wounded human peritoneal mesothelial cells (HPMC) were treated, at pH 7.4, with either (1) control medium (M199), (2) laboratory-prepared heat or filter-sterilized solutions, (3) 10% to 80% vol/vol solution of Gambrosol or Gambrosol-trio (1.5% and 4.0% glucose), (4) D-glucose (5 to 80 mmol/L), or (5) individual or combined GDP [acetaldehyde, formaldehyde, glyoxal, methylglyoxal, 3-deoxyglucosone (3-DG), 5-hydroxy methylfufural (5-HMF), or 3,4-di-deoxyglucosone-3-ene (3,4-DGE)]. Wound closure was recorded by time-lapse photomicroscopy. RESULTS: In untreated HPMC, the rate of wound closure was linear and the process was complete by 18.4 +/- 3.6 hours (N = 16). In wounded HPMC exposed to dilutions of heat-sterilized but not filtered laboratory solutions (1.5% or 4.0% glucose, pH 7.4), remesothelialization was significantly retarded (P = 0.04 and P = 0.009 vs. M199, respectively). In Gambrosol, remesothelialization was significantly retarded in both 1.5% and 4.0% solutions. In contrast in Gambrosol-trio-treated HPMC, this rate was not significantly reduced in either 1.5% or 4.0% glucose peritoneal dialysis fluids. Remesothelialization was dose-dependently retarded in HPMC exposed to 3,4-DGE (>10 microl/L), formaldehyde (>5 micromol/L) but not by exposure to the other GDP tested even at 5 times the concentration present in low glucose solutions. The rate of remesothelialization was not significantly altered by exposure to D-glucose concentrations up to 80 mmol/L. CONCLUSION: These data identify that the formaldehyde and 3,4-DGE present in heat-sterilized peritoneal dialysis solutions are important in reducing mesothelial cell regeneration. Specifically targeting their removal may have major benefits in preserving the mesothelium during long-term peritoneal dialysis.

Secretion of oncostatin M by infiltrating neutrophils: regulation of IL-6 and chemokine expression in human mesothelial cells.

Recently, we identified that regulation of leukocyte recruitment by IL-6 requires shedding of the IL-6R from infiltrating neutrophils. In this study, experiments have examined whether other IL-6-related cytokines possess similar properties. Levels of oncostatin M (OSM) and leukemia inhibitory factor were analyzed in patients with overt bacterial peritonitis during the first 5 days of infection. Although no change in leukemia inhibitory factor was observed throughout the duration of infection, OSM was significantly elevated on day 1 and rapidly returned to baseline by days 2-3. The source of OSM was identified as the infiltrating neutrophils, and OSM levels correlated both with leukocyte numbers and i.p. soluble IL-6R (sIL-6R) levels. FACS analysis revealed that OSM receptor beta expression was restricted to human peritoneal mesothelial cells. Stimulation of human peritoneal mesothelial cells with OSM induced phosphorylation of gp130 and OSM receptor beta, which was accompanied by activation of STAT3 and secretion of CC chemokine ligand 2/monocyte chemoattractant protein-1 and IL-6. Although OSM itself did not modulate CXC chemokine ligand 8/IL-8 release, it effectively suppressed IL-1beta-mediated expression of this neutrophil-activating CXC chemokine. Moreover, OSM synergistically blocked IL-1beta-induced CXC chemokine ligand 8 secretion in combination with the IL-6/sIL-6R complex. Thus suggesting that OSM and sIL-6R release from infiltrating neutrophils may contribute to the temporal switch between neutrophil influx and mononuclear cell recruitment seen during acute inflammation.