Splenic Immune Suppression in Sepsis: A Role for IL-10-Induced Changes in P38 MAPK Signaling

doi:10.1006/jsre.1998.5556

Journal of Surgical Research

Volume 83, Issue 1, 1 May 1999, Pages 36-43

Presented at the Annual Meeting of the Association for Academic Surgery, Seattle, Washington, November 18–22, 1998

https://doi.org/10.1006/jsre.1998.5556 Get rights and content

Abstract

Background.Studies have indicated that following the induction of sepsis, there is a late (24 h) generalized suppression of the immune response which is associated with increased anti-inflammatory mediator release (e.g., IL-10). However, the mechanisms by which this occurs are unknown. In this regard, recent studies indicate that p38 mitogen-activated protein kinase (p38 MAPK) may play a central role in transducing the signals from immunosuppressive agents which in turn may alter lymphoid cytokine release. The aim of this study, therefore, was to determine whether the anti-inflammatory mediator IL-10 alters splenocyte IL-2 and IFN-γ release, as well as the expression and activation of p38 MAPK in septic animals.

Materials and methods.Splenocytes (SPL) (or for some experiments purified T cells) were harvested from mice subjected 24 h earlier to either sepsis by cecal ligation and puncture (CLP) or Sham-CLP and stimulated with 2.5 μg concanavalin A (ConA)/ml in the presence or absence of either monoclonal antibody (Mab) to IL-10 (4 μg/ml) or IgG control. In subsequent studies, sepsis was induced in C57BL/6J and C57BL/6 IL-10 knockout mice, and SPL harvested and stimulated with ConA. SPL cytokine release was measured by ELISA, and the expression and phosphorylation of p38 MAPK were measured by Western analysis.

Results.The results indicate that Th1 cytokine (IL-2, IFN-γ) release was depressed by sepsis, while p38 MAPK expression and activity were increased in SPL as well as in T-cells. Neutralization of IL-10 byin vitrouse of anti-IL-10 Mab and in the IL-10 knockout animal restored the Th1 response and caused a downregulation of p38 MAPK expression and activity after CLP. Thus, IL-10 appears to contribute to the increase in p38 MAPK activity and expression and the corresponding suppression of Th1 response seen in late sepsis.

References (33)

A. Ayala et al.
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.
(1994)
J.L. Kelly et al.
Anti-interleukin-10 antibody restores burn-induced defects in T-cell function
Surgery
(1997)
S. Matsuda et al.
T lymphocyte activation signals for interleukin-2 production involve activation of MKK6-p38 and MKK7-SAPK/JNK signaling pathways sensitive to cyclosporin A
J. Biol. Chem.
(1998)
H. Niiro et al.
MAP kinase pathways as a route for regulatory mechanisms of IL-10 and IL-4 which inhibit COX-2 expression in human monocytes
Biochem. Biophys. Res. Commun.
(1998)
D.R. Meldrum et al.
Diltiazem restores IL-2, IL-3, IL-6 and IFN-gamma synthesis and decreases susceptibility to sepsis following hemorrhage
J. Surg. Res.
(1991)
J.B. Crawley et al.
T cell proliferation in response to interleukins 2 and 7 requires p38 MAP kinase activation
J. Biol. Chem.
(1997)
Z. Guan et al.
Induction of cyclooxgenase-2 by the activated MEKK1-SEK1/MKK4- p38 mitogen-activated protein kinase pathway
J. Biol. Chem.
(1998)
R.J.A. Goris
Sepsis and multiple organ failure: The result of whole body inflammation
Host Defense Dysfunction in Trauma, Shock and Sepsis
(1993)
J.A. Mannick
Trauma, sepsis and immune defects
A.E. Baue
Multiple Organ Failure. Patient Care and Prevention
(1990)

T.R. Mosmann et al.

TH1 and TH2 cells: Different patterns of lymphokine secretion lead to different functional properties

Annu. Rev. Immunol.

(1989)

S.T. O'Sullivan et al.

Major injury leads to predominance of the T helper-2 lymphocyte phenotype and diminished interleukin-12 production associated with decreased resistance to infection

Ann. Surg.

(1995)

J.T. DiPiro et al.

Association of interleukin-4 plasma levels with traumatic injury and clinical course

Arch. Surg.

(1998)

M. Howard et al.

Biological properties of interleukin 10

J. Clin. Immunol.

(1992)

C.H. Selzman et al.

Therapeutic implications of interleukin-10 in surgical disease

Shock

(1998)

A. Ayala et al.

Mechanism of enhanced susceptibility to sepsis following hemorrhage: Interleukin (IL)-10 suppression of T-cell response is mediated by eicosanoid induced IL-4 release

Arch. Surg.

(1994)

Cited by (30)

Cecal ligation and puncture: The gold standard model for polymicrobial sepsis?
2011, Trends in Microbiology
Citation Excerpt :
This is exemplified by neutralization of endogenously produced IL-10 which resulted in increased plasma levels of TNF and enhanced mortality [45]. However, a delayed therapy with anti-IL-10 monoclonal antibodies was successful after CLP-induced sepsis [46]. This might be explained by the contribution of the endogenous anti-inflammatory reaction to the initiation of immunosuppression.
Sepsis is a serious medical condition characterized by dysregulated systemic inflammatory responses followed by immunosuppression. To study the pathophysiology of sepsis, diverse animal models have been developed. Polymicrobial sepsis induced by cecal ligation and puncture (CLP) is the most frequently used model because it closely resembles the progression and characteristics of human sepsis. Here we summarize the role of several immune components in the pathogenesis of sepsis induced by CLP. However, several therapies proposed on the basis of promising results obtained by CLP could not be translated to the clinic. This demonstrates that experimental sepsis models do not completely mimic human sepsis. We propose several strategies to narrow the gap between experimental sepsis models and clinical sepsis, including targeting factors that contribute to the immunosuppressive phase of sepsis, and reproducing the heterogeneity of human patients.
Treatment with histone deacetylase inhibitor attenuates MAP kinase mediated liver injury in a lethal model of septic shock
2010, Journal of Surgical Research
Citation Excerpt :
Inducers of IL-10 synthesis include pro-inflammatory cytokines, such as TNF-α and IL-1 [34]. IL-10, in turn, can activate [48] or suppress [35, 49, 50] the p38 pathway. In our pretreatment model, the decrease in p38 and the altered transcription of the cytokine genes (decreased IL-6 and increased IL-10) may be a direct effect of the SAHA treatment, or a reflection of an overall attenuation of the inflammatory response.
Despite global efforts to improve the treatment of sepsis, it remains a leading cause of morbidity and mortality in intensive care units. We have previously shown that suberoylanilide hydroxamic acid (SAHA), a histone deacetylase inhibitor, markedly improves survival in a murine model of lipopolysaccharide (LPS)-induced shock. SAHA has anti-inflammatory properties that have not been fully characterized. The liver plays an important role in the production of acute phase reactants involved in the inflammatory cascade and is also one of the major organs that can become dysfunctional in septic shock. The purpose of this study was to assess the effect of SAHA treatment on MAP kinases and associated inflammatory markers in murine liver after LPS-induced injury.
C57B1/6J mice were randomly divided into three groups: (A) experimental-given intraperitoneal (i.p.) SAHA (50 mg/kg) in dimethyl sulfoxide (DMSO) vehicle solution (n = 12); (B) control- given vehicle only (n = 12), and; (C) sham-given no treatment (n = 7). Two hours later, experimental and control mice were injected with LPS (20 mg/kg, i.p.) and experimental mice received a second dose of SAHA. Livers were harvested at 3, 24, and 48 h for analysis of inflammatory markers using Western Blot, Polymerase Chain Reaction (PCR), and Enzyme-Linked Immunosorbent Assay (ELISA) techniques.
After 3 h, the livers of animals treated with SAHA showed significantly (P < 0.05) decreased expression of the pro-inflammatory MAP kinases phosphorylated p38, phosphorylated ERK, myeloperoxidase and interleukin-6, and increased levels of the anti-inflammatory interleukin-10 compared with controls. Phospho-p38 expression remained low in the SAHA treated groups at 24 and 48 h.
Administration of SAHA is associated with attenuation of MAPK activation and alteration of inflammatory and anti-inflammatory markers in murine liver after a lethal LPS insult. The suppression of MAPK activity is rapid (within 3 h), and is sustained for up to 48 h post-treatment. These results may in part account for the improvement in survival shown in this model.
NF-κB and P38 MAPK Inhibition Improve Survival in Endotoxin Shock and in a Cecal Ligation and Puncture Model of Sepsis in Combination With Antibiotic Therapy
2009, Journal of Surgical Research
Citation Excerpt :
Endotoxin or lipopolysaccharide (LPS) released by gram-negative bacteria also plays a major role in the pathogenesis of sepsis by inducing the release of a diverse range of proinflammatory mediators associated with bacterial sepsis [4]. The outcome from sepsis and its associated syndromes in the surgical intensive care unit has changed little despite major advances in antibiotic therapy, operative intervention, hyperalimentation, and invasive anesthesia [5]. Furthermore, there is also growing concern that antibiotic mediated endotoxin release in both sepsis and trauma may be a contributing factor in lethality from sepsis [6–8].
Nuclear factor-kappa B (NF-κB) and p38 mitogen-activated protein kinase (MAPK) are critical intracellular signal transduction pathways that mediate the systemic inflammatory response syndrome. Antibiotics induce bacterial lysis, which also contributes to cytokine production and the inflammatory response by activating NF-κB and p38 kinase. In this study, we set out to examine the effects of inhibition of p38 MAPK and NF-κB translation in in vivo models of sepsis.
Intraperitoneal lipopolysaccharide (30 mg/kg), tail vein injection of bacteria (Staphylococcus aureus + Salmonella Typhimurium, 5 × 10⁷ colony forming units/kg) and cecal ligation and puncture (CLP) with or without antibiotics (Augmentin, 100 mg/kg) were the septic models used. Animals received control, SB-202190 (a p38 inhibitor), or SN-50 (an NF-κB inhibitor), and mortality was assessed by log-rank analysis. Blood was collected at different time points for cytokine analysis, and splenic tissue was used for cytoplasmic protein extraction to assess kinase activation.
SB-202190 and SN-50 resulted in significant survival benefit in the lipopolysaccharide model (P = 0.0006) but not bacterial or CLP models (P = 0.9 and 0.3, respectively). SB-202190 and SN-50, in combination with antibiotic, resulted in a significant survival benefit in the CLP model (P = 0.0001 and 0.006, respectively). Circulating levels of both tumor necrosis factor-alpha and interleukin-6 were significantly reduced at 2 h (P = 0.047 and 0.036, respectively) and Western blot demonstrated down-regulation of p38 kinase 2 h after CLP in animals treated with p38MAPK and SN-50 inhibitors in combination with antibiotics.
We have demonstrated that p-38 and NF-κB inhibition improve survival in endotoxin shock, whereas the survival benefit in polymicrobial sepsis requires coexistent antibiotic treatment.
Combined alcohol and burn injury differentially regulate P-38 and ERK activation in mesenteric lymph node T cell
2004, Journal of Surgical Research
Recent studies from our laboratory have suggested that acute alcohol ingestion prior to burn injury enhances gut bacterial translocation by suppressing T cell-mediated intestinal immune defense. To determine the mechanism responsible for suppressed T cell function, we examined the activation of mitogen-activated protein kinases (MAPK) members p-38 and ERK-1/2. Both p-38 and ERK-1/2 are known to play a significant role in the T cell proliferation and their cytokine production. Rats were gavaged with ethanol to achieve a blood alcohol level of approximately 100 mg/dl, before they were subjected to a 25% total body surface area burn injury. Two days after injury, rats were sacrificed and mesenteric lymph node (MLN) T cells were isolated and their ability to proliferate in response to anti-CD3 was determined. For p-38 and ERK-1/2 determination, T cells were divided into two groups. Cells in one group were stimulated with anti-CD3 for 3 min and lysed. The cells in the second group were cultured for ∼18 h in the presence of anti-CD3 and lysed. MAPK status in 18-h cultured cells allowed us to determine whether or not the changes in p-38 and ERK-1/2 are transient or persist in the proliferating cells. Two days after injury, anti-CD3-mediated MLN T cell proliferation was more suppressed in rats gavaged with alcohol prior to burn injury compared to rats receiving either burn injury alone or sham-injured rats regardless of their exposure. Western blot analyses showed significant inhibition of ERK-1/2 phosphorylation in both freshly isolated and 18-h cultured T cells from alcohol and burn-injured rats compared to the sham rat T cells. The inhibition of p-38 phosphorylation in T cells derived from alcohol and burn-injured rats was found to be transient as no significant difference in p-38 phosphorylation was noted between the 18 h incubated MLN T cells of sham and alcohol and burn-injured rats. Taken together, our findings suggest that low levels of ERK-1/2 activation is likely to play a significant role in MLN T cell proliferative suppression in alcohol and burn-injured rats.
Interleukin-10 and the regulation of mitogen-activated protein kinases: Are these signalling modules targets for the anti-inflammatory action of this cytokine?
2003, Cellular Signalling
The many specific, yet overlapping and redundant activities of individual cytokines have been the basis for current concepts of therapeutical intervention. Cytokines are powerful two-edged weapons that can trigger a cascade of reactions and may show activities that often go beyond the single highly specific property that it is hoped they possess. Nevertheless, it can be stated that our new, though burgeoning, understanding of the biological mechanisms governing cytokine actions is an important contribution to medical knowledge. The crucial role of the anti-inflammatory cytokine, interleukin (IL)-10, in regulating potential molecular pathway mediating injury and cell death has attracted paramount attention in recent years. In this respect, the mitogen-activated protein kinase (MAPK) components have emerged as potential signalling cascades that regulate a plethora of cell functions, including inflammation and cell death. The biochemistry and molecular biology of cytokine actions, particularly IL-10, explain some well known and sometimes also some of the more obscure clinical aspects of the evolution of diseases.
IL-10 mediation of activation-induced Th<inf>1</inf> cell apoptosis and lymphoid dysfunction in polymicrobial sepsis
2001, Cytokine
Recent studies suggest that increased activation-induced lymphocyte apoptosis (AICD) is detected in mouse splenocytes during polymicrobial sepsis which may contribute to lymphocyte immune dysfunction [i.e., decreased interleukin (IL-)2 and interferon-γ (IFN-γ) production] leading to the associated morbidity seen in those animals. Thus, we wanted to examine the hypothesis that immune suppressive agents, such as IL-4, IL-10 or prostaglandin E₂(PGE₂), known to be elevated in septic animals, also contribute to this increase in AICD. Here we demonstrate that the inclusion of monoclonal antibody (mAb) to IL-10, but not anti-IL-4 or ibuprofen (IBU), blunted this sepsis induced increase in splenocyte AICD. Additionally, septic mice deficient in the IL-10 gene product (−/−) showed neither an increase in AICD nor a loss of IL-2/IFN-γ release capacity. Interestingly, mAb to IL-10 did not altered the extent of AICD in a Th₂-cell line, but exogenous IL-10 did potentiate Th₁-like cell line AICD. This was consistent with the finding that the increased AICD seen in septic mouse splenocytes was restricted largely to the CD4⁺cells producing IL-2 (Th₁-cells) and that mAb to IL-10 treatment suppressed this change. Furthermore, IL-10 appears to mediate its AICD effect by upregulation of the Fas receptor and Fas receptor signaling protein components, but not by altered expression ofBcl/Bax/Bad family members, in septic mouse splenocytes. To the extent that these processes contribute in a pathological fashion to the animal's capacity to survive sepsis we have previously observed that in vivo post-treatment of mice with mAb IL-10 markedly attenuated septic mortality. Collectively, these data indicate that in the septic mouse the Th₂cytokine IL-10 not only serves to actively induce Th₁lymphocyte immune dysfunction but also plays a role in their apoptotic depletion. These processes in turn appear to contribute to the animal's inability to ward off lethal septic challenge.

View all citing articles on Scopus

^☆: E. FaistJ. MeakinsF. W. Schildberg

¹: To whom correspondence and reprint requests should be addressed at Center for Surgical Research, Middle House 2, Rhode Island Hospital, 593 Eddy Street, Providence, RI 02903. Fax: (401) 444-3278. E-mail:[email protected].

View full text

Regular ArticleSplenic Immune Suppression in Sepsis: A Role for IL-10-Induced Changes in P38 MAPK Signaling☆

Abstract

J. Surg. Res.

Surgery

J. Biol. Chem.

Biochem. Biophys. Res. Commun.

J. Surg. Res.

J. Biol. Chem.

J. Biol. Chem.

Sepsis and multiple organ failure: The result of whole body inflammation

Host Defense Dysfunction in Trauma, Shock and Sepsis

Trauma, sepsis and immune defects

Multiple Organ Failure. Patient Care and Prevention

TH1 and TH2 cells: Different patterns of lymphokine secretion lead to different functional properties

Annu. Rev. Immunol.

Major injury leads to predominance of the T helper-2 lymphocyte phenotype and diminished interleukin-12 production associated with decreased resistance to infection

Ann. Surg.

Association of interleukin-4 plasma levels with traumatic injury and clinical course

Arch. Surg.

Biological properties of interleukin 10

J. Clin. Immunol.

Therapeutic implications of interleukin-10 in surgical disease

Shock

Mechanism of enhanced susceptibility to sepsis following hemorrhage: Interleukin (IL)-10 suppression of T-cell response is mediated by eicosanoid induced IL-4 release

Arch. Surg.

Regular Article
Splenic Immune Suppression in Sepsis: A Role for IL-10-Induced Changes in P38 MAPK Signaling☆