IL-22 and IL-22-Binding Protein Are Associated With Development of and Mortality From Acute-on-Chronic Liver Failure - PubMed (original) (raw)
. 2019 Jan 17;3(3):392-405.
doi: 10.1002/hep4.1303. eCollection 2019 Mar.
Affiliations
- PMID: 30859151
- PMCID: PMC6396350
- DOI: 10.1002/hep4.1303
IL-22 and IL-22-Binding Protein Are Associated With Development of and Mortality From Acute-on-Chronic Liver Failure
Katharina Schwarzkopf et al. Hepatol Commun. 2019.
Abstract
Interleukin-22 (IL-22) has context-dependent hepatoprotective or adverse properties in vitro and in animal models. IL-22 binding protein (IL-22BP) is a soluble inhibitor of IL-22 signaling. The role of IL-22 and IL-22BP in patients with acute-on-chronic liver failure (ACLF) is unclear. Beginning in August 2013, patients with liver cirrhosis with and without ACLF were prospectively enrolled and followed at predefined time points. IL-22 and IL-22BP concentrations were quantified and associated with clinical endpoints. The impact of IL-22BP on hepatocellular IL-22 signaling was assessed by functional experiments. A total of 139 patients were analyzed, including 45 (32%), 52 (37%), and 42 (30%) patients with compensated/stable decompensated liver cirrhosis, acute decompensation of liver cirrhosis, and ACLF at baseline, respectively. Serum levels of IL-22 and IL-22BP were strongly associated with the presence of, or progression to, ACLF (P < 0.001), and with mortality (P < 0.01). Importantly, the mean IL-22BP levels exceeded IL-22 levels more than 300-fold. Furthermore, IL-22BP/IL-22 ratios were lowest in patients with adverse outcomes (i.e., ACLF and death). In vitro experiments showed that IL-22BP at these concentrations inhibits hepatocellular IL-22 signaling, including the induction of acute-phase proteins. The capacity of patient serum to induce signal transducer and activator of transcription 3 phosphorylation was substantially higher in the presence of low versus high IL-22BP/IL-22 ratios. Conclusion: Our study reveals that high IL-22 levels and low ratios of IL-22BP/IL-22 are associated with ACLF and mortality of patients with cirrhosis. Excessive secretion of IL-22BP can neutralize IL-22 in vitro and may prevent-likely in a context-specific manner-hepatoprotective, but also adverse effects, of IL-22 in patients with cirrhosis.
Figures
Figure 1
IL‐22 serum concentrations are associated with adverse outcomes of liver cirrhosis. (A) IL‐22 serum concentrations are shown according to the stage of liver cirrhosis. (B) Receiver operating characteristic analysis for IL‐22 serum concentration for the presence of ACLF. An IL‐22 serum concentration of 47.68 pg/mL has been identified as an optimal cutoff to predict ACLF. (C) IL‐22 serum concentration according to the grade of ACLF, as defined by the CLIF‐EASL consortium. (D) Association between serum concentration of IL‐22 and short‐term outcome of patients with liver cirrhosis. IL‐22 concentrations were quantified at baseline and grouped according to the respective outcomes until day 28 of follow‐up as follows: No ACLF: patients without ACLF at baseline and without progression to ACLF until follow‐up; progression (prog.) to ACLF: patients without ACLF at baseline but with progression to ACLF until follow‐up; regression (regr.) of ACLF: patients with ACLF at baseline but with resolution of ACLF until follow‐up; stable ACLF: patients with persistent ACLF from baseline until follow‐up. (E) IL‐22 concentrations were quantified at baseline and compared between patients who died or who survived until day 28 of follow‐up. Abbreviations: AUC, area under the curve; comp., compensated; decomp., decompensated; gr., grade; and ROC, receiver operating characteristic.
Figure 2
IL‐22 serum concentrations are associated with infections in patients without ACLF. (A) IL‐22 serum concentrations were quantified at baseline in patients without (A) or with (B) ACLF and compared within these groups according to the presence or absence of infection, as indicated.
Figure 3
IL‐17A serum concentrations are not associated with ACLF. (A) IL‐17A serum concentrations are shown according to the indicated stage of liver cirrhosis. (B) IL‐17A serum concentrations are shown according to the grade of ACLF, as defined by the CLIF‐EASL consortium. (C) IL‐17A concentrations were quantified at baseline and compared between patients who died or who survived until day 28 of follow‐up. Abbreviations: comp., compensated; decomp., decompensated; and gr., grade.
Figure 4
Liver cirrhosis stage‐dependent excess of IL‐22BP. (A) IL‐22BP serum concentrations are shown according to the indicated stage of liver cirrhosis. (B) The ratio of serum concentrations of IL‐22BP divided through serum concentrations of IL‐22 (shown in Fig. 1) were calculated and compared among patients with the indicated stages of liver cirrhosis. Significantly lower ratios of IL‐22BP/IL‐22 are observed in patients with decompensated liver cirrhosis and ACLF compared with patients with compensated liver cirrhosis or healthy controls. (C) IL‐22BP concentrations were quantified at baseline and compared between patients who died or who survived until day 28 of follow‐up. (D) The ratio of serum concentrations of IL‐22BP divided through serum concentrations of IL‐22 were calculated and compared between patients who died or who survived until day 28 of follow‐up. (E) The ratio of IL‐22BP to IL‐22 serum concentration is associated with liver synthesis capacity. The ratio of serum concentrations of IL‐22BP divided through serum concentrations of IL‐22 were calculated in all patients with liver cirrhosis and correlated with INR (left panel) and with serum albumin concentration (right panel). Abbreviations: comp., compensated; decomp., decompensated; and gr., grade.
Figure 5
IL‐22BP inhibits hepatocellular IL‐22 signaling. (A) Expression of IL‐22Rα on Huh‐7 and HepG2 cells. IL‐22Rα was labeled with AF488 and nuclei were stained with DAPI (4´,6‐diamidino‐2‐phenylindole). (B) IL‐22 induces Stat1 and Stat3 phosphorylation in Huh‐7 and HepG2 cells. (C) IL‐22BP inhibits IL‐22‐induced Stat1 and Stat3 phosphorylation at physiological dose ranges. (D) IL‐22BP inhibits IL‐22 downstream signaling in primary human hepatocytes. Primary human hepatocytes were stimulated with or without 5 ng/mL of IL‐22 and ascending concentrations of IL‐22BP for 15 minutes, as indicated, and cellular phospho‐Stat3, total Stat3, and β‐actin protein levels were quantified by immunoblot analysis.
Figure 6
Ability of patient serum to induced hepatocellular IL‐22 signaling depends on the IL‐22BP/IL‐22 ratio. Huh‐7 cells were incubated with representative serum samples of patients with low and high IL‐22BP/IL‐22 ratios for 15 minutes. (A) Western blot analyses for the indicated antigens were performed. IL‐22BP and IL‐22 concentrations as well as IL‐22BP/IL‐22 ratios of the patient samples are shown. (B) Cells were pretreated with or without a neutralizing antibody against the IL‐22 receptor before exposure to patient serum with high IL‐22 activity (1 and 2 from [A]) for 15 minutes.
Figure 7
Effect of IL‐22BP on IL‐22‐induced production of proinflammatory mediators and cell viability. (A) Cell viability was assessed in Huh‐7 cells using the WST‐1 assay 6 hours after stimulation with IL‐22 with or without IL‐22BP at the indicated concentrations. (B) Induction of TNF‐α mRNA in Huh‐7.5 cells treated for 6 hours with the indicated concentrations of IL‐22. TNF‐α mRNA levels were quantified by quantitative polymerase chain reaction (PCR) and expressed relative to GAPDH (glyceraldehyde 3‐phosphate dehydrogenase). (C) Huh‐7 cells and HepG2 cells were stimulated with IL‐22 with or without IL‐22BP at the indicated concentrations for 6 hours. mRNA levels of LBP were quantified by quantitative PCR and expressed relative to GAPDH. (D) Quantification of LBP protein levels in HepG2 cells stimulated with IL‐22 for the indicated time points. (E) IL‐22BP abolishes IL‐22‐mediated LBP protein production in HepG2 cells and primary human hepatocytes. Cells were stimulated with 5 ng/mL of IL‐22 and ascending concentrations of IL‐22BP for 24 hours. Protein levels were analyzed by immunoblotting using β‐actin as a control.
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