Disordered macrophage cytokine secretion underlies impaired acute inflammation and bacterial clearance in Crohn's disease - PubMed (original) (raw)
. 2009 Aug 31;206(9):1883-97.
doi: 10.1084/jem.20091233. Epub 2009 Aug 3.
Farooq Z Rahman, Bu'Hussain Hayee, Simon J Graham, Daniel J B Marks, Gavin W Sewell, Christine D Palmer, Jonathan Wilde, Brian M J Foxwell, Israel S Gloger, Trevor Sweeting, Mark Marsh, Ann P Walker, Stuart L Bloom, Anthony W Segal
Affiliations
- PMID: 19652016
- PMCID: PMC2737162
- DOI: 10.1084/jem.20091233
Disordered macrophage cytokine secretion underlies impaired acute inflammation and bacterial clearance in Crohn's disease
Andrew M Smith et al. J Exp Med. 2009.
Erratum in
- J Exp Med. 2009 Sep 28;206(10):2301
Abstract
The cause of Crohn's disease (CD) remains poorly understood. Counterintuitively, these patients possess an impaired acute inflammatory response, which could result in delayed clearance of bacteria penetrating the lining of the bowel and predispose to granuloma formation and chronicity. We tested this hypothesis in human subjects by monitoring responses to killed Escherichia coli injected subcutaneously into the forearm. Accumulation of (111)In-labeled neutrophils at these sites and clearance of (32)P-labeled bacteria from them were markedly impaired in CD. Locally increased blood flow and bacterial clearance were dependent on the numbers of bacteria injected. Secretion of proinflammatory cytokines by CD macrophages was grossly impaired in response to E. coli or specific Toll-like receptor agonists. Despite normal levels and stability of cytokine messenger RNA, intracellular levels of tumor necrosis factor (TNF) were abnormally low in CD macrophages. Coupled with reduced secretion, these findings indicate accelerated intracellular breakdown. Differential transcription profiles identified disease-specific genes, notably including those encoding proteins involved in vesicle trafficking. Intracellular destruction of TNF was decreased by inhibitors of lysosomal function. Together, our findings suggest that in CD macrophages, an abnormal proportion of cytokines are routed to lysosomes and degraded rather than being released through the normal secretory pathway.
Figures
Figure 1.
Neutrophil accumulation and subsequent clearance of E. coli from the tissues is markedly delayed in a dose-dependent manner in CD. 111Indium-labeled autologous neutrophils were injected intravenously at the same time as killed E. coli were injected subcutaneously into each forearm. (a) Radioactivity measured over the injection sites showed a much smaller proportion of labeled cells accumulating in CD subjects. (b) γ-Camera image of a CD patient at 24 h after injection, demonstrating focal accumulations of radioactivity at bacterial injection sites (arrows) and confirming lack of bowel inflammation. (c) 32P-labeled killed E. coli were injected into the subcutaneous tissues of the forearm and radioactivity was measured at the skin surface. Clearance of radioactivity was much slower in CD than in HC or UC. Extrapolating these curves indicated that almost complete removal (99%) would take 10.2 and 7.1 d in HC and UC subjects, respectively, compared with 44.3 d in CD. (d and e) Effect of increasing bacterial dose from 105 to 108 on blood flow at injection site (d) and bacterial clearance (e). The numbers of subjects studied in the dose response experiment are depicted in e. All results are expressed as mean ± SEM (**, P < 0.01; ***, P < 0.001).
Figure 2.
Proinflammatory cytokine secretion by macrophages from CD patients is deficient in response to E. coli. Supernatants from macrophages stimulated for 24 h with HkEc were tested for the levels of cytokines and chemokines. (a) Macrophages from HC subjects released varying amounts of cytokines and chemokines after HkEc stimulation. (b) Cytokine and chemokine release expressed as a percentage of that secreted by HC cells (blue bar) from ileal and colonic CD patients. The numbers of subjects in each group are shown on left; each patient was used once. All results are expressed as mean ± SEM (*, P < 0.05; **, P < 0.01; ***, P < 0.001).
Figure 3.
Abnormal transcriptional profiles of CD macrophages on Affymetrix gene arrays. (a) Venn diagram shows the number of probe sets differing in expression (P < 0.01) between ileal and colonic CD versus HC macrophages in the unstimulated state and after exposure to HkEc. (b and c) As in a, but comparing differentially expressed probe sets between ileal and colonic CD and UC versus HC in unstimulated (b) and HkEc-stimulated (c) cells. (d) i-iv designate overlaps of differentially expressed genes of known function after HkEc stimulation between disease groups, as labeled in Venn diagrams.
Figure 4.
Intracellular levels of TNF are lower in CD, despite normal transcription and mRNA stability, but restored to normal in the presence of Bref-A and lysosome inhibitors. (a) Macrophages transfected with an adenoviral vector containing a TNF promoter and luciferase reporter demonstrated equivalent TNF transcription levels in HC (n = 8) and CD (n = 7) subjects. (b) TNF mRNA stability is comparable in HC and CD macrophages after stimulation with HkEc. (c) Intracellular levels of TNF after HkEc stimulation with or without Bref-A were determined by Western blotting. (d) CD contained significantly lower levels of intracellular TNF than HC macrophages after HkEc stimulation but returned to normal levels with the inclusion of Bref-A (n = 6 in all groups). (e) Intracellular levels of TNF after HkEc stimulation with or without lysosomal inhibitors were determined by Western blotting. (f) Inhibitors of lysosomal proteolysis increase intracellular TNF levels in HC and CD macrophages (n = 4 in all groups; gray line denotes level of HkEc alone). Significance levels are compared with HkEc alone. All patients in these studies had colonic CD and subjects were used once per assay. Results are shown as mean ± SEM (*, P < 0.05).
Figure 5.
Intracellular levels of chemokines and cytokines after HkEc stimulation in the presence of vesicle trafficking and lysosomal inhibitors. (a) Intracellular cytokine array profiles obtained from macrophages stimulated with HkEc in the presence of absence of either Bref-A or monensin. (b) Cytokine array map. (c) Various chemokines and cytokines demonstrate reduced intracellular levels in CD macrophages after HkEc stimulation and are either normalized or elevated after Bref-A or monensin treatment (n = 4 in all groups). All patients studied had colonic CD and subjects were used once per assay. Results shown are mean ± SEM (*, P < 0.05; ***, P < 0.001).
Comment in
- The 2-phase model of Crohn's disease: from immune defect to hyperresponse.
Salas A, Panés J. Salas A, et al. Gastroenterology. 2010 Mar;138(3):1204-6; discussion 1207. doi: 10.1053/j.gastro.2010.01.025. Epub 2010 Jan 25. Gastroenterology. 2010. PMID: 20100444 No abstract available.
Similar articles
- Optineurin deficiency in mice contributes to impaired cytokine secretion and neutrophil recruitment in bacteria-driven colitis.
Chew TS, O'Shea NR, Sewell GW, Oehlers SH, Mulvey CM, Crosier PS, Godovac-Zimmermann J, Bloom SL, Smith AM, Segal AW. Chew TS, et al. Dis Model Mech. 2015 Aug 1;8(8):817-29. doi: 10.1242/dmm.020362. Epub 2015 Jun 4. Dis Model Mech. 2015. PMID: 26044960 Free PMC article. - Monocyte-derived macrophages from Crohn's disease patients are impaired in the ability to control intracellular adherent-invasive Escherichia coli and exhibit disordered cytokine secretion profile.
Vazeille E, Buisson A, Bringer MA, Goutte M, Ouchchane L, Hugot JP, de Vallée A, Barnich N, Bommelaer G, Darfeuille-Michaud A. Vazeille E, et al. J Crohns Colitis. 2015 May;9(5):410-20. doi: 10.1093/ecco-jcc/jjv053. Epub 2015 Mar 24. J Crohns Colitis. 2015. PMID: 25805890 - Defective tumor necrosis factor release from Crohn's disease macrophages in response to Toll-like receptor activation: relationship to phenotype and genome-wide association susceptibility loci.
Sewell GW, Rahman FZ, Levine AP, Jostins L, Smith PJ, Walker AP, Bloom SL, Segal AW, Smith AM. Sewell GW, et al. Inflamm Bowel Dis. 2012 Nov;18(11):2120-7. doi: 10.1002/ibd.22952. Epub 2012 Mar 20. Inflamm Bowel Dis. 2012. PMID: 22434667 Free PMC article. - The role of neutrophils in the pathogenesis of Crohn's disease.
Segal AW. Segal AW. Eur J Clin Invest. 2018 Nov;48 Suppl 2:e12983. doi: 10.1111/eci.12983. Eur J Clin Invest. 2018. PMID: 29931668 Review. - What is wrong with granulocytes in inflammatory bowel diseases?
Levine AP, Segal AW. Levine AP, et al. Dig Dis. 2013;31(3-4):321-7. doi: 10.1159/000354686. Epub 2013 Nov 14. Dig Dis. 2013. PMID: 24246982 Free PMC article. Review.
Cited by
- Monocyte-macrophages modulate intestinal homeostasis in inflammatory bowel disease.
Lu H, Suo Z, Lin J, Cong Y, Liu Z. Lu H, et al. Biomark Res. 2024 Aug 2;12(1):76. doi: 10.1186/s40364-024-00612-x. Biomark Res. 2024. PMID: 39095853 Free PMC article. Review. - Elucidating the role of diet in maintaining gut health to reduce the risk of obesity, cardiovascular and other age-related inflammatory diseases: recent challenges and future recommendations.
Aziz T, Hussain N, Hameed Z, Lin L. Aziz T, et al. Gut Microbes. 2024 Jan-Dec;16(1):2297864. doi: 10.1080/19490976.2023.2297864. Epub 2024 Jan 4. Gut Microbes. 2024. PMID: 38174551 Free PMC article. Review. - Cutaneous manifestations of inflammatory bowel disease: basic characteristics, therapy, and potential pathophysiological associations.
He R, Zhao S, Cui M, Chen Y, Ma J, Li J, Wang X. He R, et al. Front Immunol. 2023 Oct 26;14:1234535. doi: 10.3389/fimmu.2023.1234535. eCollection 2023. Front Immunol. 2023. PMID: 37954590 Free PMC article. Review. - Metabolic and functional remodeling of colonic macrophages in response to high-fat diet-induced obesity.
Castoldi A, Sanin DE, van Teijlingen Bakker N, Aguiar CF, de Brito Monteiro L, Rana N, Grzes KM, Kabat AM, Curtis J, Cameron AM, Caputa G, Antônio de Souza T, Souto FO, Buescher JM, Edwards-Hicks J, Pearce EL, Pearce EJ, Saraiva Camara NO. Castoldi A, et al. iScience. 2023 Aug 25;26(10):107719. doi: 10.1016/j.isci.2023.107719. eCollection 2023 Oct 20. iScience. 2023. PMID: 37674984 Free PMC article. - Predicting diagnostic biomarkers associated with immune infiltration in Crohn's disease based on machine learning and bioinformatics.
Bao W, Wang L, Liu X, Li M. Bao W, et al. Eur J Med Res. 2023 Jul 26;28(1):255. doi: 10.1186/s40001-023-01200-9. Eur J Med Res. 2023. PMID: 37496049 Free PMC article.
References
- Aggarwal B.B., Kohr W.J., Hass P.E., Moffat B., Spencer S.A., Henzel W.J., Bringman T.S., Nedwin G.E., Goeddel D.V., Harkins R.N. 1985. Human tumor necrosis factor. Production, purification, and characterization.J. Biol. Chem. 260:2345–2354 - PubMed
- Ambrose N.S., Alexander-Williams J., Keighley M.R. 1984a. Audit of sepsis in operations for inflammatory bowel disease.Dis. Colon Rectum. 27:602–604 - PubMed
- Ambrose N.S., Johnson M., Burdon D.W., Keighley M.R. 1984b. Incidence of pathogenic bacteria from mesenteric lymph nodes and ileal serosa during Crohn's disease surgery.Br. J. Surg. 71:623–625 - PubMed
- Barrett J.C., Hansoul S., Nicolae D.L., Cho J.H., Duerr R.H., Rioux J.D., Brant S.R., Silverberg M.S., Taylor K.D., Barmada M.M., et al. ; NIDDK IBD Genetics Consortium; Belgian-French IBD Consortium; Wellcome Trust Case Control Consortium 2008. Genome-wide association defines more than 30 distinct susceptibility loci for Crohn's disease.Nat. Genet. 40:955–962 - PMC - PubMed
- Bazzoni F., Beutler B. 1996. The tumor necrosis factor ligand and receptor families.N. Engl. J. Med. 334:1717–1725 - PubMed
Publication types
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
Other Literature Sources
Medical