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Molecular Medicine

, Volume 14, Issue 5–6, pp 247–256 | Cite as

Diversity of Interferon γ and Granulocyte-Macrophage Colony-Stimulating Factor in Restoring Immune Dysfunction of Dendritic Cells and Macrophages During Polymicrobial Sepsis

  • Stefanie B. Flohé
  • Hemant Agrawal
  • Sascha Flohé
  • Meenakshi Rani
  • Jörg M. Bangen
  • F. Ulrich Schade
Research Article

Abstract

The development of immunosuppression during polymicrobial sepsis is associated with the failure of dendritic cells (DC) to promote the polarization of T helper (Th) cells toward a protective Th1 type. The aim of the study was to test potential immunomodulatory approaches to restore the capacity of splenic DC to secrete interleukin (IL) 12 that represents the key cytokine in Th1 cell polarization. Murine polymicrobial sepsis was induced by cecal ligation and puncture (CLP). Splenic DC were isolated at different time points after CLP or sham operation, and stimulated with bacterial components in the presence or absence of neutralizing anti-IL-10 antibodies, murine interferon (IFN) γ, and/or granulocyte macrophage colony-stimulating factor (GM-CSF). DC from septic mice showed an impaired capacity to release the pro-inflammatory and Th1-promoting cytokines tumor necrosis factor α, IFN-γ, and IL-12 in response to bacterial stimuli, but secreted IL-10. Endogenous IL-10 was not responsible for the impaired IL-12 secretion. Up to 6 h after CLP, the combined treatment of DC from septic mice with IFN-γ and GM-CSF increased the secretion of IL-12. Later, DC from septic mice responded to IFN-γ and GM-CSF with increased expression of the co-stimulatory molecule CD86, while IL-12 secretion was no more enhanced. In contrast, splenic macrophages from septic mice during late sepsis responded to GM-CSF with increased cytokine release. Thus, therapy of sepsis with IFN-γ/GM-CSF might be sufficient to restore the activity of macrophages, but fails to restore DC function adequate for the development of a protective Th1-like immune response.

Notes

Acknowledgments

This work is supported by DFG grant FL-353/2-1 (to Stefanie B Flohé). We are grateful to Michaela Bak for excellent technical assistance and to Ernst Kreuzfelder and to Bärbel Nyadu for support in flow cytometry.

References

  1. 1.
    Angus DC, Linde-Zwirble WT, Lidicker J, Clermont G, Carcillo J, Pinsky MR. (2001) Epidemiology of severe sepsis in the United States: analysis of incidence, outcome, and associated costs of care. Crit. Care Med. 29:1303–10.CrossRefPubMedGoogle Scholar
  2. 2.
    Munoz C, Carlet J, Fitting C, Misset B, Bleriot JP, Cavaillon JM. (1991) Dysregulation of in vitro cytokine production by monocytes during sepsis. J. Clin. Invest. 88:1747–54.CrossRefPubMedPubMedCentralGoogle Scholar
  3. 3.
    Ayala A, Chaudry IH. (1996) Immune dysfunction in murine polymicrobial sepsis: mediators, macrophages, lymphocytes and apoptosis. Shock. 6 Suppl. 1:S27–38.CrossRefGoogle Scholar
  4. 4.
    Ferguson NR, Galley HF, Webster NR. (1999) T helper cell subset ratios in patients with severe sepsis. Intensive Care Med. 25:106–9.CrossRefPubMedGoogle Scholar
  5. 5.
    Heidecke CD, et al. (1999) Selective defects of T lymphocyte function in patients with lethal intraabdominal infection. Am. J. Surg. 178:288–92.CrossRefGoogle Scholar
  6. 6.
    Ayala A, Deol ZK, Lehman DL, Herdon CD, Chaudry IH. (1994) Polymicrobial sepsis but not low-dose endotoxin infusion causes decreased splenocyte IL-2/IFN-gamma release while increasing IL-4/IL-10 production. J. Surg. Res. 56:579–85.CrossRefPubMedGoogle Scholar
  7. 7.
    Banchereau J, Steinman RM. (1998) Dendritic cells and the control of immunity. Nature. 392:245–52.CrossRefGoogle Scholar
  8. 8.
    Edwards AD, et al. (2002) Microbial recognition via Toll-like receptor-dependent and -independent pathways determines the cytokine response of murine dendritic cell subsets to CD40 triggering. J. Immunol. 169:3652–60.CrossRefPubMedGoogle Scholar
  9. 9.
    Cella M, Scheidegger D, Palmer-Lehmann K, Lane P, Lanzavecchia A, Alber G. (1996) Ligation of CD40 on dendritic cells triggers production of high levels of interleukin-12 and enhances T cell stimulatory capacity: T-T help via APC activation. J. Exp. Med. 184:747–52.CrossRefPubMedGoogle Scholar
  10. 10.
    Aste-Amezaga M, Ma X, Sartori A, Trinchieri G. (1998) Molecular mechanisms of the induction of IL-12 and its inhibition by IL-10. J. Immunol. 160:5936–44.PubMedGoogle Scholar
  11. 11.
    Corinti S, Albanesi C, la Sala A, Pastore S, Girolomoni G. (2001) Regulatory activity of autocrine IL-10 on dendritic cell functions. J. Immunol. 166:4312–8.CrossRefPubMedGoogle Scholar
  12. 12.
    Boonstra A, et al. (2006) Macrophages and myeloid dendritic cells, but not plasmacytoid dendritic cells, produce IL-10 in response to MyD88- and TRIF-dependent TLR signals, and TLR-independent signals. J. Immunol. 177:7551–8.CrossRefPubMedGoogle Scholar
  13. 13.
    Scumpia PO, et al. (2005) CD11c+ dendritic cells are required for survival in murine polymicrobial sepsis. J. Immunol. 175:3282–6.CrossRefPubMedGoogle Scholar
  14. 14.
    Hotchkiss RS, et al. (2002) Depletion of dendritic cells, but not macrophages, in patients with sepsis. J. Immunol. 168:2493–500.CrossRefGoogle Scholar
  15. 15.
    Hiramatsu M, Hotchkiss RS, Karl IE, Buchman TG. (1997) Cecal ligation and puncture (CLP) induces apoptosis in thymus, spleen, lung, and gut by an endotoxin and TNF-independent pathway. Shock. 7:247–53.CrossRefPubMedGoogle Scholar
  16. 16.
    Efron PA, et al. (2004) Characterization of the systemic loss of dendritic cells in murine lymph nodes during polymicrobial sepsis. J. Immunol. 173:3035–43.CrossRefPubMedGoogle Scholar
  17. 17.
    Tinsley KW, et al. (2003) Sepsis induces apoptosis and profound depletion of splenic interdigitating and follicular dendritic cells. J. Immunol. 171:909–14.CrossRefPubMedGoogle Scholar
  18. 18.
    Guisset O, et al. (2007) Decrease in circulating dendritic cells predicts fatal outcome in septic shock. Intensive Care Med. 33:148–52.CrossRefGoogle Scholar
  19. 19.
    Flohé SB, Agrawal H, Schmitz D, Gertz M, Flohé S, Schade FU. (2006) Dendritic cells during polymicrobial sepsis rapidly mature but fail to initiate a protective Th1-type immune response. J. Leukoc. Biol. 79:473–81.CrossRefPubMedGoogle Scholar
  20. 20.
    Benjamim CF, Lundy SK, Lukacs NW, Hogaboam CM, Kunkel SL. (2005) Reversal of long-term sepsis-induced immunosuppression by dendritic cells. Blood. 105:3588–95.CrossRefPubMedPubMedCentralGoogle Scholar
  21. 21.
    Moreno SE, Alves-Filho JC, Alfaya TM, da Silva JS, Ferreira SH, Liew FY. (2006) IL-12, but not IL-18, is critical to neutrophil activation and resistance to polymicrobial sepsis induced by cecal ligation and puncture. J. Immunol. 177:3218–24.CrossRefPubMedGoogle Scholar
  22. 22.
    Sparwasser T, et al. (1997) Bacterial DNA causes septic shock. Nature. 386:336–7.CrossRefPubMedGoogle Scholar
  23. 23.
    Deng J, Muthu K, Gamelli R, Shankar R, Jones SB. (2004) Adrenergic modulation of splenic macrophage cytokine release in polymicrobial sepsis. Am. J. Physiol. Cell Physiol. 287:C730–6.CrossRefPubMedGoogle Scholar
  24. 24.
    Ding Y, et al. (2004) Polymicrobial sepsis induces divergent effects on splenic and peritoneal dendritic cell function in mice. Shock. 22:137–44.CrossRefPubMedPubMedCentralGoogle Scholar
  25. 25.
    Emmanuilidis K, et al. (2001) Critical role of Kupffer cell-derived IL-10 for host defense in septic peritonitis. J. Immunol. 167:3919–27.CrossRefPubMedGoogle Scholar
  26. 26.
    Yang S, Koo DJ, Zhou M, Chaudry IH, Wang P. (2000) Gut-derived norepinephrine plays a critical role in producing hepatocellular dysfunction during early sepsis. Am. J. Physiol. Gastrointest. Liver Physiol. 279:G1274–81.CrossRefPubMedGoogle Scholar
  27. 27.
    Kalinski P, Hilkens CM, Wierenga EA, Kapsenberg ML. (1999) T-cell priming by type-1 and type-2 polarized dendritic cells: the concept of a third signal. Immunol. Today. 20:561–7.CrossRefGoogle Scholar
  28. 28.
    Strobl H, Knapp W. (1999) TGF-beta1 regulation of dendritic cells. Microbes. Infect. 1:1283–90.CrossRefPubMedGoogle Scholar
  29. 29.
    Maestroni GJ. (2002) Short exposure of maturing, bone marrow-derived dendritic cells to norepinephrine: impact on kinetics of cytokine production and Th development. J. Neuroimmunol. 129:106–14.CrossRefPubMedGoogle Scholar
  30. 30.
    De Smedt T, Van Mechelen M, De Becker G, Urbain J, Leo O, Moser M. (1997) Effect of interleukin-10 on dendritic cell maturation and function. Eur. J. Immunol. 27:1229–35.CrossRefPubMedGoogle Scholar
  31. 31.
    Wen H, Hogaboam CM, Gauldie J, Kunkel SL. (2006) Severe sepsis exacerbates cell-mediated immunity in the lung due to an altered dendritic cell cytokine profile. Am. J. Pathol. 168:1940–50.CrossRefPubMedPubMedCentralGoogle Scholar
  32. 32.
    Kovarik P, et al. (1999) Stress-induced phosphorylation of STAT1 at Ser727 requires p38 mitogen-activated protein kinase whereas IFN-gamma uses a different signaling pathway. Proc. Natl. Acad. Sci. U. S. A. 96:13956–61.CrossRefPubMedPubMedCentralGoogle Scholar
  33. 33.
    McLeish KR, et al. (1998) Activation of mitogen-activated protein kinase cascades during priming of human neutrophils by TNF-alpha and GM-CSF. J. Leukoc. Biol. 64:537–45.CrossRefPubMedGoogle Scholar
  34. 34.
    Rani M, Husain B, Lendemans S, Schade FU, Flohe S. (2006) Haemorrhagic shock in mice—intracellular signaling and immunomodulation of peritoneal macrophages’ LPS response. Immunobiology. 211:711–9.CrossRefPubMedGoogle Scholar
  35. 35.
    Welte T, Koch F, Schuler G, Lechner J, Doppler W, Heufler C. (1997) Granulocyte-macrophage colony-stimulating factor induces a unique set of STAT factors in murine dendritic cells. Eur. J. Immunol. 27:2737–40.CrossRefPubMedGoogle Scholar
  36. 36.
    Flohé S, et al. (1999) Influence of granulocyte-macrophage colony-stimulating factor (GM-CSF) on whole blood endotoxin responsiveness following trauma, cardiopulmonary bypass, and severe sepsis. Shock. 12:17–24.CrossRefPubMedGoogle Scholar
  37. 37.
    Williams MA, et al. (1998) Granulocyte-macrophage colony-stimulating factor induces activation and restores respiratory burst activity in monocytes from septic patients. J. Infect. Dis. 177:107–15.CrossRefPubMedGoogle Scholar
  38. 38.
    Nierhaus A, et al. (2003) Reversal of immunoparalysis by recombinant human granulocyte-macrophage colony-stimulating factor in patients with severe sepsis. Intensive Care Med. 29:646–51.CrossRefGoogle Scholar
  39. 39.
    Docke WD, et al. (1997) Monocyte deactivation in septic patients: restoration by IFN-gamma treatment. Nat. Med. 3:678–81.CrossRefGoogle Scholar
  40. 40.
    Dries DJ, et al. (1994) Effect of interferon gamma on infection-related death in patients with severe injuries. A randomized, double-blind, placebo-controlled trial. Arch. Surg. 129:1031–41; discussion 1042.CrossRefGoogle Scholar
  41. 41.
    Polk HC, Jr, et al. (1992) A randomized prospective clinical trial to determine the efficacy of interferon-gamma in severely injured patients. Am. J. Surg. 163:191–6.CrossRefGoogle Scholar
  42. 42.
    Mels AK, et al. (2001) Immune-stimulating effects of low-dose perioperative recombinant granulocyte-macrophage colony-stimulating factor in patients operated on for primary colorectal carcinoma. Br. J. Surg. 88:539–44.CrossRefPubMedGoogle Scholar
  43. 43.
    Rosenbloom AJ, Linden PK, Dorrance A, Penkosky N, Cohen-Melamed MH, Pinsky MR. (2005) Effect of granulocyte-monocyte colony-stimulating factor therapy on leukocyte function and clearance of serious infection in nonneutropenic patients. Chest. 127:2139–50.CrossRefGoogle Scholar
  44. 44.
    Toda H, et al. (1994) Effect of granulocyte-macrophage colony-stimulating factor on sepsis-induced organ injury in rats. Blood. 83:2893–8.PubMedGoogle Scholar
  45. 45.
    Kylanpaa ML, et al. (2005) Monocyte anergy is present in patients with severe acute pancreatitis and is significantly alleviated by granulocyte-macrophage colony-stimulating factor and interferon-gamma in vitro. Pancreas. 31:23–7.CrossRefPubMedGoogle Scholar

Copyright information

© Feinstein Institute for Medical Research 2008

Authors and Affiliations

  • Stefanie B. Flohé
    • 1
  • Hemant Agrawal
    • 1
    • 2
  • Sascha Flohé
    • 1
  • Meenakshi Rani
    • 1
  • Jörg M. Bangen
    • 1
  • F. Ulrich Schade
    • 1
  1. 1.Surgical Research, Department of Trauma Surgery, University Hospital EssenUniversity Duisburg-EssenEssenGermany
  2. 2.Arthritis and Immunology ProgramOklahoma Medical Research FoundationOklahoma CityUSA

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