Galectin-9 protects mice from the Shwartzman reaction by attracting prostaglandin E2-producing polymorphonuclear leukocytes

doi:10.1016/j.clim.2007.04.015

Clinical Immunology

Volume 124, Issue 2, August 2007, Pages 221-233

https://doi.org/10.1016/j.clim.2007.04.015 Get rights and content

Abstract

Galectins play a crucial role in the modulation of innate and adaptive immunity. Here we show that galectin-9 (Gal-9) exhibits an anti-inflammatory role in LPS-induced inflammation. Intraperitoneal LPS injection enhances Gal-9 levels as well as promotes the production of pro-inflammatory cytokines, e.g., TNF-alpha, IFN-gamma and IL-12. We found that Gal-9 administration results in the protection of mice from the Shwartzman reaction, and Gal-9-deficient mice became susceptible to the Shwartzman reaction, thus implying the anti-inflammatory activity of Gal-9 against LPS-induced inflammation. Indeed, Gal-9 treatment together with LPS suppresses production of these pro-inflammatory cytokines, while it rather enhances than suppresses IL-4 and IL-10 production. We also found that LPS-induced elevation of TNF-alpha, IFN-gamma, and IL-12 does not occur in Gal-9 transgenic mice. Moreover, Gal-9 induces Gr-1⁺ cell; probably polymorphonuclear leukocyte (PMN), as well as infiltration in to the peritoneal cavity, causing us to hypothesize PMNs are involved in Gal-9-mediated suppression. The fact that Gal-9 does not suppress LPS-induced TNF-alpha, IFN-gamma and IL-12 production in neutropenic mice, and that it does not protect those mice from the Shwartzman reaction, confirms the involvement of PMN in regulation. PMN attracted by Gal-9 produce PGE₂, which LPS-induced TNF-alpha production from the peritoneal macrophages is suppressed, while PMNs attracted by casein produce less PGE₂ and fail to suppress LPS-induced TNF-alpha production. Our data suggest that Gal-9 regulates LPS-induced inflammation and protects mice from the Shwartzman reaction by attracting PGE₂-producing PMN.

Introduction

Galectins are soluble metal-independent lectins bound to β-galactoside-containing glycoconjugates exhibiting a variety of biological activities, such as cell adhesion, proliferation [1], [2], [3], apoptosis [4], [5], and cell-cycle progression [6]. Galectins are expressed by a variety of immune and inflammatory cells besides epithelial cells and exhibit pleiotropic functions such as pro- and anti-inflammatory functions [7], [8], [9].

Galectin-9 (Gal-9) has been originally classified as an eosinophil chemoattractant, inducing superoxide production and prolonging cell survival [10], [11]. More recently, it has been shown that Gal-9 negatively regulates Th1 type immunity as a Tim-3 ligand [12]. We have also shown that Gal-9 expression is up-regulated by IFN-γ and IL-1β in various cell types [1], [13], [14], and that it induces maturation of immature dendritic cells (DC), suggesting that Gal-9 plays a crucial role in both innate and adaptive immunity [15]. As noted, Gal-1, Gal-3 and Gal-8 exhibit several important functions in PMN, the effector cells in the host's immune response against bacterial infection and inflammation, such as induction of superoxide and adhesion to endothelial cells [16], [17], [18], [19], [20], [21], [22]. Polymorphonuclear leukocytes (PMNs) have, thus, been long believed to play a major regulatory role in infectious disease by producing cytokines and chemokines, such as TNF-α, IL-1β, IL-8, IL-10, GM-CSF, RANTES, and MIP-1α by LPS stimulation [23].

Recently, it has been shown that there are several subsets of PMN through the analysis of their macrophage (Mϕ) modification activities [24], and that resident PMN may convert to pro-inflammatory or anti-inflammatory PMN in response to host circumstances. Furthermore, it has been shown that PMN produce an unidentified factor that inhibits the release of TNF-α and IL-6 from activated Mϕ [25], suggesting that PMNs are not a single and terminally differentiated population, but have a subset that also suppresses inflammation through Mϕ modification. The mechanistic details of this have yet to be clarified.

The purpose of the present investigation is to show a novel function of Gal-9 in the regulation of pro-inflammatory cytokine production in innate immunity.

Section snippets

Mice

Male BALB/c and C57BL/6 mice were purchased from Japan SLC. BALB/c, C57BL/6, Gal-9-transgenic (BALB/c) and Gal-9-deficient (C57BL/6) mice were maintained on a 12:12-h light–dark cycle in a conventional animal facility at the Kagawa University. The animals were fed a standard laboratory diet and water ad libitum. All experimental procedures were approved by the Animal Care and Use Committee, conforming to the Guidelines for Animal Experimentation, Kagawa University.

Induction of LPS-induced inflammation and lethal Shwartzman reaction

Male BALB/c mice (8 weeks old,

Up-regulation of Gal-9 level in LPS-induced inflammation

First, we assessed Gal-9 levels in PLF during LPS-induced peritoneal inflammation. Fig. 1A illustrates that Gal-9 was detectable in PLF even before LPS injection. The Gal-9 levels increased after LPS injection, peaking at 2 to 12 h, and returning to baseline 48 h after injection. Furthermore, we found that higher Gal-9 levels in PLF were observed in Gal-9-TG mice compared to wild-type (WT) mice during LPS-induced stimulation (Fig. 1B). These results raise the possibility that Gal-9 plays a role

Discussion

Here we describe a novel function of Gal-9, suppression of LPS-induced pro-inflammatory cytokine production, such as TNF-α, IL-12 and IFN-γ by attracting PGE₂-producing PMN, resulting in the protection of mice from the Shwartzman reaction. We have previously shown contradictory results whereby Gal-9 induced maturation of immature DC that secrete IL-12, resulting in enhanced TNF-α and IFN-γ cytokine production [15]. We also observed DC stimulation by low-dose Gal-9, enough for DC maturation and

Acknowledgments

We greatly appreciate and thank Dr. S. Narumiya for his helpful discussions and donation of PGE₂ antagonists.

This study was in part supported by a grant from the Japanese Ministry of Education, Culture, Sports, Science, and Technology.

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Citation Excerpt :
What's more, pckGal-9 induced the expression of anti-inflammatory cytokine— IL-10, initially known as cytokine synthesis inhibitory factor, is a multifunctional cytokine and demonstrated immunosuppressive function [41]. Likewise, a previous literature had found that Galectin-9 regulates LPS-induced inflammation and protects mice from shwartzman reaction, indicating the crucial role of Galectin-9 in bacterial infection [42]. Taken together, our results agreed well with the role of Galectin-9 in mediating pro-inflammatory responses.
Galectins are β-galactoside sugar binding proteins which function as important pattern recognition receptors (PRRs) in innate immunity. Here, we identified a galectin-9 gene from koi carp (Cyprinus carpio), named kGal-9. The ORF of kGal-9 is 963 bp in length, which encodes a polypeptide of 320 amino acids without either signal peptide. The predicted molecular weight is 36.25 kDa, and the isoelectric point is 8.3. Multiple sequence alignment showed that the putative kGal-9 contains two carbohydrate recognition domains (CRD), which are conserved in Galectin-9s. The phylogenetic tree showed that kGal-9 clustered to Galectin-9s from other teleosts, and shared the highest identity of 87.5% with Qihe crucian (Carassius auratus). kGal-9 mRNA was abundant in head kidney, gills, and gut, but low in liver and muscle. Further, the expression level of kGal-9 in the head kidney and liver increased significantly after Aeromonas veronii (abbreviated A.v) infection. Unexpectedly, kGal-9 showed a remarkable downregulation in the spleen at various time points post A.v infection. Intramuscular injection of pckGal-9 not merely reduced the bacterial load of spleen tissue, but also improved the survival rate of koi carp post A.v challenge. Besides, administration of pckGal-9 stimulated the expression of several immuno-related genes including proinflammatory cytokines (IL-1β, IL-6), anti-inflammatory cytokine (IL-10), complement components (C4, C9), with fluctuation in spleen and head kidney. Taken together, the obtained results suggest that kGal-9 occupies an important role in innate immunity and defense against bacterial infection in koi carp.
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¹: Y.T. and H.A. contributed equally to this work.

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Galectin-9 protects mice from the Shwartzman reaction by attracting prostaglandin E2-producing polymorphonuclear leukocytes

Abstract

Introduction

Section snippets

Mice

Induction of LPS-induced inflammation and lethal Shwartzman reaction

Up-regulation of Gal-9 level in LPS-induced inflammation

Discussion

Acknowledgments

J. Biol. Chem.

Clin. Immunol.

Trends Immunol.

J. Biol. Chem.

Blood

Immunol. Today

Immunity

J. Biol. Chem.

Blood

Selective eosinophil adhesion to fibroblast via IFN-gamma-induced galectin-9

J. Immunol.

Galectins: versatile modulators of cell adhesion, cell proliferation, and cell death

J. Mol. Med.

Role of galectin-8 as a modulator of cell adhesion and cell growth

Glycoconjugate J.

Galectin-9 induces apoptosis through the calcium-calpain-caspase-1 pathway

J. Immunol.

Apoptosis of T cells mediated by galectin-1

Nature

Shedding light on the immunomodulatory properties of galectins: novel regulators of innate and adaptive immune responses

Glycoconjugate J.

Biological activities of ecalectin: a novel eosinophil-activating factor

J. Immunol.

The Tim-3 ligand galectin-9 negatively regulates T helper type 1 immunity

Nat. Immunol.

Interferon-γ stimulates the expression of galectin-9 in cultured human endothelial cells

J. Leukocyte Biol.

Interleukin-1beta stimulates galectin-9 expression in human astrocytes

NeuroReport

Galectin-9 induces maturation of human monocyte-derived dendritic cells

J. Immunol.

Activation of the neutrophil nicotinamide adenine dinucleotide phosphate oxidase by galectin-1

J. Immunol.

Galectin-9 protects mice from the Shwartzman reaction by attracting prostaglandin E₂-producing polymorphonuclear leukocytes