Aryl Hydrocarbon Receptor Ligands in Cigarette Smoke Induce Production of Interleukin-22 to Promote Pancreatic Fibrosis in Models of Chronic Pancreatitis

doi:10.1053/j.gastro.2016.09.064

Gastroenterology

Volume 151, Issue 6, December 2016, Pages 1206-1217

https://doi.org/10.1053/j.gastro.2016.09.064 Get rights and content

Background & Aims

Cigarette smoke has been identified as an independent risk factor for chronic pancreatitis (CP). Little is known about the mechanisms by which smoking promotes development of CP. We assessed the effects of aryl hydrocarbon receptor (AhR) ligands found in cigarette smoke on immune cell activation in humans and pancreatic fibrosis in animal models of CP.

Methods

We obtained serum samples from patients with CP treated at Stanford University hospital and healthy individuals (controls) and isolated CD4⁺ T cells. Levels of interleukin-22 (IL22) were measured by enzyme-linked immunosorbent assay and smoking histories were collected. T cells from healthy nonsmokers and smokers were stimulated and incubated with AhR agonists (2,3,7,8-tetrachlorodibenzo-p-dioxin or benzo[a]pyrene) or antagonists and analyzed by flow cytometry. Mice were given intraperitoneal injections of caerulein or saline, with or without lipopolysaccharide, to induce CP. Some mice were given intraperitoneal injections of AhR agonists at the start of caerulein injection, with or without an antibody against IL22 (anti-IL22) starting 2 weeks after the first caerulein injection, or recombinant mouse IL22 or vehicle (control) intraperitoneally 4 weeks after the first caerulein injection. Mice were exposed to normal air or cigarette smoke for 6 h/d for 7 weeks and expression of AhR gene targets was measured. Pancreata were collected from all mice and analyzed by histology and quantitative reverse transcription polymerase chain reaction. Pancreatic stellate cells and T cells were isolated and studied using immunoblot, immunofluorescence, flow cytometry, and enzyme-linked immunosorbent analyses.

Results

Mice given AhR agonists developed more severe pancreatic fibrosis (based on decreased pancreas size, histology, and increased expression of fibrosis-associated genes) than mice not given agonists after caerulein injection. In mice given saline instead of caerulein, AhR ligands did not induce fibrosis. Pancreatic T cells from mice given AhR agonists and caerulein were activated and expressed IL22, but not IL17 or interferon gamma. Human T cells exposed to AhR agonists up-regulated expression of IL22. In mice given anti-IL22, pancreatic fibrosis did not progress, whereas mice given recombinant IL22 had a smaller pancreas and increased fibrosis. Pancreatic stellate cells isolated from mouse and human pancreata expressed the IL22 receptor IL22RA1. Incubation of the pancreatic stellate cells with IL22 induced their expression of the extracellular matrix genes fibronectin 1 and collagen type I α1 chain, but not α2 smooth muscle actin or transforming growth factor−β. Serum samples from smokers had significantly higher levels of IL22 than those from nonsmokers.

Conclusions

AhR ligands found in cigarette smoke increase the severity of pancreatic fibrosis in mouse models of pancreatitis via up-regulation of IL22. This pathway might be targeted for treatment of CP and serve as a biomarker of disease.

Section snippets

Mice and Treatments

All animal protocols were approved by Institutional Animal Care and Use Committees of Stanford University. All mice including Balb/c, C57BL/6J, and AhR^d were purchased from Jackson Laboratory (Bar Harbor, ME) and were housed under pathogen-free conditions. Experimental mice were age- and sex-matched. Bone marrow (BM) chimeric mice were generated by lethally irradiating mice with 9.5 Gy γ-radiation in 2 doses approximately 3 hours apart, followed by intravenous injection of 5 × 10⁶ BM cells from

Aryl Hydrocarbon Receptor Activation Worsens Fibrosis in Chronic Pancreatitis

Cigarette smoking is an independent risk factor for accelerating CP,6, 9 however, the mechanism remains elusive. Cigarette smoke contains AhR agonists, such as dioxin and BaP10, 11; in addition, cigarette smoke was found to have an unexpectedly high dioxin-like potential that triggers AhR activation.¹² Therefore, we sought to investigate the role of cigarette smoke AhR ligands on immune activation and on the pathogenesis of CP.

The well-characterized potent AhR agonist TCDD was administrated to

Discussion

CP is a complex chronic inflammation disorder with links to genetic, metabolic, and environmental factors. Alcohol is the most established environmental risk factor for CP, while the independent effects and risks associated with smoking have not been elucidated until recently. In most populations, smoking is strongly associated with drinking alcohol,²⁵ therefore, the independent effect of smoking can be difficult to assess. However, data from case-control²⁶ and population-based studies5, 27

Acknowledgments

The authors thank Wenjun Ouyang from Genentech for providing anti-IL22 antibody. The authors thank Yi Wei for technical assistance.

Jing Xue and Qinglan Zhao contributed equally to this work. Walter Park and Aida Habtezion contributed equally to this work.

References (42)

H. Witt et al.
Chronic pancreatitis: challenges and advances in pathogenesis, genetics, diagnosis, and therapy
Gastroenterology
(2007)
H.Y. Gaisano et al.
New insights into the mechanisms of pancreatitis
Gastroenterology
(2009)
M.M. Lerch et al.
Models of acute and chronic pancreatitis
Gastroenterology
(2013)
J. Xue et al.
Aryl hydrocarbon receptor regulates pancreatic IL-22 production and protects mice from acute pancreatitis
Gastroenterology
(2012)
T. Zelante et al.
Tryptophan catabolites from microbiota engage aryl hydrocarbon receptor and balance mucosal reactivity via interleukin-22
Immunity
(2013)
H. Goecke et al.
Macrophages infiltrating the tissue in chronic pancreatitis express the chemokine receptor CCR5
Surgery
(2000)
J. Mimura et al.
Functional role of AhR in the expression of toxic effects by TCDD
Biochim Biophys Acta
(2003)
Y. Nagashio et al.
Inhibition of transforming growth factor beta decreases pancreatic fibrosis and protects the pancreas against chronic injury in mice
Lab Invest
(2004)
S.J. Rulyak et al.
Risk factors for the development of pancreatic cancer in familial pancreatic cancer kindreds
Gastroenterology
(2003)
M.V. Apte et al.
A starring role for stellate cells in the pancreatic cancer microenvironment
Gastroenterology
(2013)

C.E. Forsmark

The early diagnosis of chronic pancreatitis

Clin Gastroenterol Hepatol

(2008)

H. Sarles

Etiopathogenesis and definition of chronic pancreatitis

Dig Dis Sci

(1986)

D. Yadav et al.

The role of alcohol and smoking in pancreatitis

Nat Rev Gastroenterol Hepatol

(2010)

V.W. Setiawan et al.

Prospective study of alcohol drinking, smoking, and pancreatitis: the multiethnic cohort

Pancreas

(2016)

P. Maisonneuve et al.

Cigarette smoking accelerates progression of alcoholic chronic pancreatitis

Gut

(2005)

A. Andriulli et al.

Smoking as a cofactor for causation of chronic pancreatitis: a meta-analysis

Pancreas

(2010)

A.T. Lee et al.

Alcohol and cigarette smoke components activate human pancreatic stellate cells: implications for the progression of chronic pancreatitis

Alcohol Clin Exp Res

(2015)

J.S. Tolstrup et al.

Smoking and risk of acute and chronic pancreatitis among women and men: a population-based cohort study

Arch Intern Med

(2009)

H. Muto et al.

Dioxins in cigarette smoke

Arch Environ Health

(1989)

R.L. Stedman

The chemical composition of tobacco and tobacco smoke

Chem Rev

(1968)

A. Kasai et al.

High levels of dioxin-like potential in cigarette smoke evidenced by in vitro and in vivo biosensing

Cancer Res

(2006)

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: Conflicts of interest The authors disclose no conflicts.

: Funding This work was supported by the National Pancreas Foundation Grant, National Institute of Health (NIH) grants DK092421 and DK105263 (Aida Habtezion); National Natural Science Foundation of China grant, China State Key Laboratory of Oncogenes and Related Gene (no. 91-15-15), Program for Professor of Special Appointment (Eastern Scholar) at Shanghai Institutions of Higher Learning (no. TP2015007) (Jing Xue); Department of Veterans Affairs, and NIH CA163200 (Stephen J. Pandol), AA019996 and AA011999 (Stephen J. Pandol and Mouad Edderkaoui).

: Author names in bold designate shared co-first authorship.

^∗: Authors share co-first authorship.

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Original ResearchFull Report: Basic and Translational—PancreasAryl Hydrocarbon Receptor Ligands in Cigarette Smoke Induce Production of Interleukin-22 to Promote Pancreatic Fibrosis in Models of Chronic Pancreatitis

Background & Aims

Methods

Results

Conclusions

Section snippets

Mice and Treatments

Aryl Hydrocarbon Receptor Activation Worsens Fibrosis in Chronic Pancreatitis

Discussion

Acknowledgments

Gastroenterology

Gastroenterology

Gastroenterology

Gastroenterology

Immunity

Surgery

Biochim Biophys Acta

Lab Invest

Gastroenterology

Gastroenterology

Clin Gastroenterol Hepatol

Etiopathogenesis and definition of chronic pancreatitis

Dig Dis Sci

The role of alcohol and smoking in pancreatitis

Nat Rev Gastroenterol Hepatol

Prospective study of alcohol drinking, smoking, and pancreatitis: the multiethnic cohort

Pancreas

Cigarette smoking accelerates progression of alcoholic chronic pancreatitis

Gut

Smoking as a cofactor for causation of chronic pancreatitis: a meta-analysis

Pancreas

Alcohol and cigarette smoke components activate human pancreatic stellate cells: implications for the progression of chronic pancreatitis

Alcohol Clin Exp Res

Smoking and risk of acute and chronic pancreatitis among women and men: a population-based cohort study

Arch Intern Med

Dioxins in cigarette smoke

Arch Environ Health

The chemical composition of tobacco and tobacco smoke

Chem Rev

High levels of dioxin-like potential in cigarette smoke evidenced by in vitro and in vivo biosensing

Cancer Res

Original Research
Full Report: Basic and Translational—Pancreas
Aryl Hydrocarbon Receptor Ligands in Cigarette Smoke Induce Production of Interleukin-22 to Promote Pancreatic Fibrosis in Models of Chronic Pancreatitis