Leukocyte and endothelial adhesion molecule studies in knockout mice

doi:10.1016/j.coph.2004.01.003

Current Opinion in Pharmacology

Volume 4, Issue 2, April 2004, Pages 154-158

https://doi.org/10.1016/j.coph.2004.01.003 Get rights and content

Abstract

Ischemia and reperfusion of the myocardium initiate an inflammatory response directed against the myocardium, and many studies attribute a significant portion of this injury to leukocytes. Leukocyte and endothelial cell adhesion molecules are responsible for neutrophil–endothelial cell interactions in coronary vasculature following ischemia and reperfusion. Interactions between β₂-integrins and intercellular adhesion molecule-1 are responsible for firm adhesion of neutrophils to the coronary endothelium in acute cardiac inflammation. Leukocyte-expressed CD18 plays a crucial role, and genetic deficiency of CD18 significantly attenuates myocardial ischemia-reperfusion injury. Genetic deficiency of intercellular adhesion molecule-1 also minimizes myocardial necrosis following ischemia and reperfusion. The selectin family of adhesion glycoproteins also participates in various phases of leukocyte–endothelial interactions, and studies with P-selectin- and E-selectin-deficient mice have shown attenuation of both neutrophil accumulation and myocardial injury following myocardial ischemia and reperfusion.

Introduction

Coronary heart disease (CHD) remains the single leading cause of human mortality. According to the American Heart Association, nearly 500 000 Americans die each year from CHD, accounting for 20% of all deaths nationwide. Over one million Americans will have a recurrent coronary attack or develop a new episode this year. Accordingly, intensive investigative efforts are directed toward enhancing our understanding of heart disease and ultimately developing effective therapies to combat the deleterious consequences of CHD.

Ischemia-reperfusion (I-R) of the myocardium initiates an inflammatory response directed against the myocardium [1]. This sequence of events damages multiple cardiac cell types, resulting in profound contractile dysfunction and even death. A significant portion of this injury is attributed to leukocytes; however, acceptance of this idea is not universal. Several animal studies have provided solid foundation for the concept that neutrophils contribute to myocardial I-R injury. Current efforts have been directed at identification of the cellular and molecular determinants of neutrophil-mediated infiltration of the myocardium following I-R. Specifically, these studies address which leukocyte and endothelial cell adhesion molecules are responsible for neutrophil recruitment in the coronary vasculature. This review addresses the role of leukocyte–endothelial cell interactions in the development of myocardial injury subsequent to experimental myocardial I-R in gene-targeted mice that are deficient in various leukocyte or endothelial cell adhesion proteins.

Section snippets

Leukocyte β₂-integrins in myocardial ischemia-reperfusion injury

Many studies of myocardial I-R injury have focused on one or more of the three steps in leukocyte sequestration: rolling, firm adhesion and transmigration (Figure 1). Of these three key steps, firm adhesion has received greatest attention from both basic scientists and clinicians. We now know that adhesion of leukocytes to the vascular endothelium is largely dependent upon interactions between endothelial-bound and leukocyte-expressed adhesion molecules [2].

Leukocyte integrins are a subgroup of

Endothelial immunoglobulins in myocardial ischemia-reperfusion injury

The endothelial ligand for β₂-integrins, ICAM-1, has been extensively studied in several animal models. These studies have provided evidence of a role for ICAM-1 in neutrophil adherence. To fully elucidate the role of ICAM-1 in a variety of pathologies, genetically deficient mice have been developed [7]. We found that genetic deficiency of ICAM-1 significantly attenuated myocardial necrosis in mice in association with reductions in myocardial neutrophil infiltration, following both brief [5]

Selectins in myocardial ischemia-reperfusion injury

Endothelial selectins participate in the early (P-selectin) and late (P-selectin, E-selectin) phases of leukocyte–endothelial cell interactions (i.e. rolling) (Figure 1). P-selectin is stored as a preformed molecule in the Wiebel-Palade bodies in endothelial cells. Upon stimulation, stored P-selectin is translocated to the surface of endothelial cells within minutes. After several hours of inflammatory stimulation (e.g. myocardial I-R), de novo E-selectin and P-selectin become expressed on the

Cytokine mutant mice and myocardial reperfusion injury

Numerous cytokines, including tumour necrosis factor-α (TNF-α), interleukin (IL)-1β, IL-6 and IL-10, have been shown to initiate the inflammatory response that ensues following myocardial I-R. In contrast, IL-10 has been shown to diminish the inflammatory response in a number of disease states and is thought to be a beneficial cytokine. Interestingly, cytokines upregulate a number of endothelial cell adhesion molecules as well as activate leukocyte function [4]. A number of studies have

Conclusions

Inflammation and leukocyte infiltration subsequent to myocardial I-R clearly occur and contribute to myocardial injury in experimental models of myocardial I-R injury. Furthermore, clinical evidence exists for the involvement of leukocyte–endothelial cell interactions in the development of myocardial infarction in humans 20., 21., 22., 23.. However, questions still remain regarding the therapeutic benefit of anti-leukocyte agents in acute myocardial infarction in humans 24., 25., 26., 27., 28..

References and recommended reading

Papers of particular interest, published within the annual period of review, have been highlighted as:

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of special interest
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of outstanding interest

Acknowledgements

Experimental studies presented in this review were supported by the National Institutes of Health Grants RO1 HL-60849 and PO1 DK-43785 (to DJ Lefer).

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ROS-activated calcium signaling mechanisms regulating endothelial barrier function
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The renal injury and inflammation caused by ischemia-reperfusion are reduced by genetic inhibition of TNF-αR1: A comparison with infliximab treatment
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The role of the tumor necrosis factor (TNF)-α in the pathophysiology of renal ischemia/reperfusion (I/R) injury is unclear. We investigate the effects of TNF-αR1 gene deletion and infliximab administration on the degree of renal injury induced by I/R. TNF-αR1 knockout (TNF-αR1KO) and wild-type (TNF-αWT) mice were subjected to bilateral renal artery occlusion (30 min) and reperfusion (24 h). Infliximab (10 mg/kg subcutaneously, s.c.) was administered 1 h before ischemia. At the end of experiments, urea, creatinine, γGT, and AST were measured to assess renal function and reperfusion injury. Markers of oxidative stress, pro-inflammatory mediators, iNOS, COX-2, and NF-κB signaling pathway were measured. Kidney myeloperoxidase (MPO) activity and malondialdehyde (MDA) levels were measured to study polymorphonuclear cell infiltration and lipid peroxidation. TNF-αR1 gene deletion and infliximab administration prevented the increase of urea, creatinine, γGT, kidney AST levels, iNOS and COX-2 expression, NF-κB translocation, MPO activity and MDA levels. TNF-αR1 gene deletion and infliximab administration lowered the histological evidence of renal damage associated with I/R and caused a reduction of nitrotyrosine suggesting reduced nitrosative stress. Our results demonstrate that TNF-α plays an important role in I/R injury and put forward the hypothesis that modulation of TNF-α expression may represent a novel and possible strategy.
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Citation Excerpt :
ICAM-1 is expressed by many different cell types including EC, epithelial cells, cardiomyocytes, fibroblasts, smooth muscle cells, lymphocytes and monocytes (Benson et al., 2007; Niessen et al., 1999). In the heart, ICAM-1 has been shown to be expressed by cardiomyocytes following MI (Devaux et al., 1997; Hawkins et al., 1996; Kukielka et al., 1995b; Niessen et al., 1999) and there is good evidence that myocardial expression of ICAM-1 is detrimental in the MI setting, due to increased neutrophilic inflammation (Barnes, 2007; Graciano et al., 2001; Ioculano et al., 1994; Jones et al., 2000; Kakkar and Lefer, 2004; Lefer et al., 1996; Palazzo et al., 1998; Sato et al., 2006). The interaction between fibroblasts and neutrophils has been overlooked until recently, when it was demonstrated that neutrophils could adhere to CF or dermal fibroblasts via interactions with ICAM-1 (Couture et al., 2009).
Neutrophil attraction and adhesion to endothelial cells occurs via well defined mechanisms, yet the ability of other cell types to express neutrophil-binding adhesion molecules is not well studied. Cardiac fibroblasts (CF) are a key cell type involved in repair of the infarcted myocardium, a scenario in which neutrophil recruitment is perceived to be detrimental. Here we determined the effects of proinflammatory cytokines on expression of neutrophil-binding adhesion molecules and neutrophil-attracting chemokines in CF cultured from multiple patients, and explored the underlying regulatory mechanisms. An adhesion molecule focused RT-PCR array identified 5 transcripts that were increased markedly in human CF treated with the proinflammatory cytokine interleukin-1 (IL-1, 10 ng/ml, 6 h); including intercellular cell adhesion molecule (ICAM-1) and E-selectin. Real-time RT-PCR verified the array data and immunoblotting confirmed cytokine-induced ICAM-1 and E-selectin protein expression. Treatment with a panel of pharmacological inhibitors identified the NF-κB pathway as mediating IL-1-induced ICAM-1 and E-selectin mRNA and protein expression. Additionally, E-selectin expression in human CF was markedly potentiated by the JNK inhibitor SP600125, but this was not observed when a more selective inhibitor ((L)-JNKI-1) was used, or in human vascular endothelial cells. IL-1 also stimulated CF to secrete the neutrophil chemoattractant CXCL8 via a p38- and NF-κB-dependent mechanism, as well as inducing CXCL1, CXCL2 and CXCL5 mRNA expression. In conclusion, human CF express neutrophil-binding adhesion molecules and neutrophil chemoattractants in response to proinflammatory cytokines suggesting that, in addition to EC, CF may play an important role in regulating neutrophil recruitment into the infarcted myocardium.
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Leukocyte and endothelial adhesion molecule studies in knockout mice

Abstract

Introduction

Section snippets

Leukocyte β2-integrins in myocardial ischemia-reperfusion injury

Endothelial immunoglobulins in myocardial ischemia-reperfusion injury

Selectins in myocardial ischemia-reperfusion injury

Cytokine mutant mice and myocardial reperfusion injury

Conclusions

References and recommended reading

Acknowledgements

Pathophysiology

Cell

J Am Coll Cardiol

Am J Cardiol

Am J Cardiol

Myocardial reperfusion: a double edged sword

J Clin Invest

Modulation of leukocyte-mediated myocardial reperfusion injury

Annu Rev Physiol

Neutrophil adherence to isolated adult canine myocytes: evidence for a CD18-dependent mechanism

J Clin Invest

Inflammation in the course of early myocardial ischemia

FASEB J

Leukocyte and endothelial cell adhesion molecules in a chronic murine model of myocardial reperfusion injury

Am J Physiol

Inflammatory and immune response are impaired in mice deficient in intercellular adhesion molecule 1

Proc Natl Acad Sci U S A

Antibody to platelet/endothelial cell adhesion molecule-1 reduces myocardial infarct size in a rat model of ischemia-reperfusion injury

Circulation

Coronary endothelial cell P-selectin in the pathogenesis of ischemia-reperfusion injury

Am J Physiol

Reperfusion injury is not affected by blockade of P-selectin in the diabetic mouse heart

Am J Physiol

Leukocyte β₂-integrins in myocardial ischemia-reperfusion injury