Remote Ischemic Preconditioning Decreases Adhesion and Selectively Modifies Functional Responses of Human Neutrophils

doi:10.1016/j.jss.2008.08.010

Journal of Surgical Research

Volume 158, Issue 1, January 2010, Pages 155-161

https://doi.org/10.1016/j.jss.2008.08.010 Get rights and content

Objectives

Preconditioning of cells or organs by transient sublethal ischemia-reperfusion (IR), termed ischemic preconditioning (IPC), protects the cell or organ from a subsequent prolonged ischemic insult. The mechanisms of this effect remain to be fully elucidated. We have recently reported that IPC of a forearm results in alterations in gene expression profiles of circulating polymorphonuclear leukocytes. The goal of the current study was to determine if the observed changes in gene expression lead to functional changes in neutrophils.

Methods

We examined the effect of repetitive transient human forearm ischemia (three cycles of 5 min ischemia, followed by 5 min of reperfusion) on the function of circulating neutrophils. Neutrophil functions were examined before, after 1 d, and after 10 d of daily transient forearm ischemia. To modulate IR-induced inflammation the neutrophils were stimulated with N-formyl-methionyl-leucyl phenylalanine (FMLP) and lipopolysaccharide (LPS).

Results

Neutrophil adhesion was significantly decreased on day 1 and remained low on day 10 (P = 0.0149) without significant change in CD11b expression. Phagocytosis was significantly suppressed on day 10 compared with day 0 (P < 0.0001). Extracellular cytokine levels were low in the absence of an exogenous stimulus but stimulation with LPS induced significant changes on day 10. We observed a trend in reduction of apoptosis on day 1 and day 10 that did not reach statistical significance (P < 0.08).

Conclusion

This study indicates that repetitive IPC of the forearm results in substantial alterations in neutrophil function, including reduced adhesion, exocytosis, phagocytosis, and modified cytokine secretion.

Introduction

Ischemic preconditioning (IPC), induced by transient sublethal ischemia, protects the cell or organ from a subsequent prolonged ischemic insult [1]. More recently, transient tissue ischemia in a remote area has been shown to confer protection of another organ subsequently subjected to potentially lethal ischemia [2]. This intriguing mode of protection against ischemia-reperfusion (IR) injury is termed remote ischemic preconditioning (rIPC). We have previously shown that transient limb ischemia protects against a systemic inflammatory response consequent to cardiopulmonary bypass [3], substantially reduces the extent of myocardial infarction after coronary occlusion [4], modifies coronary blood flow and resistance [5], and reduces myocardial IR injury after heart transplantation [6] in porcine models. More recently, we have reported the first application of rIPC in patients undergoing open-heart surgery [7], with evidence of reduced cardiac damage and systemic inflammatory response in treated patients. Although the exact mechanism of the protection afforded by rIPC remains incomplete, we have also demonstrated, in humans, that the rIPC stimulus decreases expression of a portfolio of proinflammatory genes in circulating leukocytes [8].

Neutrophils play a key role in the IR injury 9, 10. Neutrophil-mediated tissue damage is dependent on the number of neutrophils infiltrating the postischemic tissue via the process known as transendothelial migration. Neutrophil transendothelial migration, in turn, is influenced by the ability of circulating neutrophils to adhere to the damaged endothelium. Neutrophil adhesion, a crucial element of IR injury, is a two-stage process of selectin-mediated loose adhesion and integrin-mediated firm adhesion. One of the key integrins in the firm adhesion is the CD11b receptor. Expression of the CD11b receptor is directly related to the extent of the post-operative IR injury 11, 12. Interestingly, in a previous experimental study, rIPC attenuated both endothelial dysfunction (abnormal flow mediated dilation) and prevented the increase in neutrophil CD11b expression in humans subjected to 40 min of forearm ischemia followed by reperfusion [13].

The aim of the current study was to determine if our previous observation of the effect of rIPC on neutrophil gene expression correlated with functional changes in the leukocytes. We therefore examined the effect of repetitive transient human forearm ischemia on selected aspects of neutrophil function.

Section snippets

Subjects and Blood Samples

We performed a longitudinal study using the rIPC protocol as previously described [13] in five healthy adult male volunteers who were taking no medications (mean age, 37.5 y). Briefly, one forearm was made ischemic by inflating a blood pressure cuff to 200 mm Hg for three 5-min periods, separated by 5 min of reperfusion. This protocol was repeated daily for 10 d. Venous blood was drawn from the contralateral arm prior to the ischemic stimulus (day 0) and on days 1 and 10 after the stimulus. All

Adhesion

Adhesion of nonstimulated neutrophils decreased from 13.0% ± 4.3% to 0.81% ± 0.2% within 24 h and remained low on day 10 (2.61% ± 0.7%); these changes were statistically significant (P < 0.015) (Fig. 1). Stimulation with FMLP mirrored the pattern observed in nonstimulated neutrophils and increased adhesion of neutrophils to 34.7% ± 4.7% in cells studied on day 0, but only to 18.6% ± 5.4% on day 1 and only to 2.9% ± 1.9% on day 10 (P = 0.0167).

CD11b Surface Expression

The surface expression of CD11b, expressed as median

Discussion

Remote ischemic preconditioning has been shown to provide important protection against IR injury in the heart and other organs in both animal models and in humans. The data from the current study suggests that part of this effect may be attributable to modulation of the inflammatory response consequent upon altered functional responsiveness of circulating leukocytes. In this regard, we have previously reported marked down-regulation of expression in a portfolio of proinflammatory genes in

Acknowledgments

This work was supported by operating grants from the Canadian Institutes of Health Research and Physicians Services Incorporated. GPD is the recipient of a Tier 1 Canada Research Chair. ANR is supported by a Transatlantic Network Grant from the Leducq Foundation.

References (30)

M.M. Cheung et al.
Randomized controlled trial of the effects of remote ischemic preconditioning on children undergoing cardiac surgery: First clinical application in humans
J Am Coll Cardiol
(2006)
G. Gross et al.
Mechanisms of postischemic contractile dysfunction
Ann Thorac Surg
(1999)
J. Park et al.
Mechanisms of myocardial reperfusion injury
Ann Thorac Surg
(1999)
C. Rinder et al.
Neutrophil CD11b upregulation during cardiopulmonary bypass is associated with postoperative renal injury
Ann Thorac Surg
(2003)
I. Vermes et al.
Flow cytometry of apoptotic cell death
J Immunol Methods
(2000)
E. Vachon et al.
CD44 is a phagocytic receptor
Blood
(2006)
Y.Y. Wang et al.
Proinflammatory cytokines may induce late preconditioning in unstable angina patients
Med Hypotheses
(2006)
B. Dawn et al.
Tumor necrosis factor-α does not modulate ischemia/reperfusion injury in naive myocardium but is essential for the development of late preconditioning
J Mol Cell Cardiol
(2004)
N. Teoh et al.
Low-dose TNF-α protects against hepatic ischemia-reperfusion injury in mice: Implications for preconditioning
Hepatology
(2003)
L.J. Eddy et al.
Tumor necrosis factor-α pretreatment is protective in a rat model of myocardial ischemia-reperfusion injury
Biochem Biophys Res Commun
(1992)

S.C. Glasgow et al.

Interleukin-1beta is prominent in the early pulmonary inflammatory response after hepatic injury

Surgery

(2005)

E.T. Clark et al.

Endothelial cell response to hypoxia-reoxygenation is mediated by IL-1

J Surg Res

(1995)

C.E. Murry et al.

Preconditioning with ischemia: A delay of lethal cell injury in ischemic myocardium

Circulation

(1986)

K. Przyklenk et al.

Regional ischemic “preconditioning” protects remote virgin myocardium from subsequent sustained coronary occlusion

Circulation

(1993)

R.K. Kharbanda et al.

Remote ischemic preconditioning protects against cardiopulmonary bypass-induced tissue injury: A preclinical study

Heart

(2006)

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Citation Excerpt :
However, the following normalization of circulating levels within 22 h may suggest that this is an event regulated in time, or even that this aggression generates a defective response, with cells presenting impaired migratory capacities. In fact, RIC alters neutrophil functional responsiveness, impairing phagocytosis, exocytosis and cytokine secretion, but also negatively modulating chemotaxis and cell adhesion (Konstantinov et al., 2004; Shimizu et al., 2010). Moreover, the assumption that the protective effect of RIC towards succeeding ischemic events also relates to the modulation of neutrophil functions, favors the growing field of innate immune training or innate immune memory in the context of both acute and chronic diseases (Kalafati et al., 2020).
Remote ischemic conditioning (RIC) is a novel promising therapy for treatment of neurological diseases, including ischemic stroke. RIC consists of short cycles of ischemia in a distant non-vital organ that may protect other organs against ischemia. Extensive experimental data and some few clinical trials support the neuroprotective role of RIC in ischemic stroke. Nevertheless, the circulating factors involved in this inter-organ communication and neuroprotection are not clarified. This pilot study in humans characterized the innate and adaptive circulating immune cell populations following RIC. This analysis has a particular focus at 24 h after RIC to avoid circadian influence. In silico functional analysis of mass spectrometry data identified 15 immune-related proteins. Our results reveal an immune response following RIC.
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VascularRemote Ischemic Preconditioning Decreases Adhesion and Selectively Modifies Functional Responses of Human Neutrophils

Objectives

Methods

Results

Conclusion

Introduction

Section snippets

Subjects and Blood Samples

Adhesion

CD11b Surface Expression

Discussion

Acknowledgments

J Am Coll Cardiol

Ann Thorac Surg

Ann Thorac Surg

Ann Thorac Surg

J Immunol Methods

Blood

Med Hypotheses

J Mol Cell Cardiol

Hepatology

Biochem Biophys Res Commun

Surgery

J Surg Res

Preconditioning with ischemia: A delay of lethal cell injury in ischemic myocardium

Circulation

Regional ischemic “preconditioning” protects remote virgin myocardium from subsequent sustained coronary occlusion

Circulation

Remote ischemic preconditioning protects against cardiopulmonary bypass-induced tissue injury: A preclinical study

Heart

Vascular
Remote Ischemic Preconditioning Decreases Adhesion and Selectively Modifies Functional Responses of Human Neutrophils