Blockade of CCL2/CCR2 signaling pathway prevents inflammatory monocyte recruitment and attenuates OVA-Induced allergic asthma in mice

doi:10.1016/j.imlet.2019.08.006

Immunology Letters

Volume 214, October 2019, Pages 30-36

https://doi.org/10.1016/j.imlet.2019.08.006 Get rights and content

Highlights

•
CCL2 and CCR2 expressions are increased in OVA mice.
•
Blockade of CCL2/CCR2 signaling suppresses Th2 cytokines and enhances Th1 cytokines.
•
Inhibition of Th2 inflammatory responses prevents allergic asthma.

Abstract

Recent studies have reported recruitment of inflammatory monocytes by cytokines including chemokine (C-C motif) ligand 2 (CCL2) are critical in allergic responses. We aimed to investigate the role of inflammatory monocytes and CCL2 in mouse model with ovalbumin (OVA)-induced allergic asthma. Mice were sensitized with OVA to induce allergic asthma. The proportion of inflammatory cells in bronchoalveolar lavage fluid (BALF) and peritoneal lavage fluid (PLF) were measured by flow cytometry. The expression of CCL2 and CCL2 receptor (CCR2) were determined by qPCR and western blot. The concentrations of Type 1 helper T (Th1) and Type 2 helper T (Th2) cytokines in PLF were detected by ELISA. Inflammatory monocytes are recruited in PLF, and expression of CCL2 and CCR2 were elevated in OVA-induced mice. In addition, transfer of CCR2 knockdown inflammatory monocytes decreased the levels of allergic asthma biomarkers. Injection of anti−CCL2 or anti−CCR2 antibody decreased the proportion of eosinophils and inflammatory monocytes in BALF. Blockade of CCL2/CCR2 signaling pathway suppressed the allergen-induced Th2 cytokines and enhanced the levels of Th1-associated cytokines. Blockade of CCL2/CCR2 signaling pathway in sensitization-recruited inflammatory monocytes exhibits protective effects in mouse model of OVA-induced allergic asthma by inhibiting the Th2 inflammatory responses.

Introduction

Bronchial asthma (referred to as asthma) is a common and frequent chronic respiratory disease affecting approximately 300 million people worldwide [1]. The main symptoms conclude paroxysmal wheezing, chest tightness, shortness of breath, and cough [2]. In recent years, the prevalence of asthma has been increasing year by year, seriously affecting the physical and mental health of patients and bringing a heavy economic and social burden to the society [3]. Therefore, in-depth research on the pathogenesis of asthma is of great significance for the control and treatment of asthma.

Allergic asthma is the most common type of asthma and is often associated with allergies and allergens [4], producing a range of allergen exposure associated symptoms including chest tightness, cough, and wheezing. It is an airway chronic inflammatory disease involving a variety of inflammatory cells (such as lymphocytes, eosinophils, mast cells, macrophages, airway epithelial cells, etc.) and inflammatory mediators [5], often characterized by the increase of biomarkers such as eosinophils, serum Immunoglobulin E (IgE), and Type 2 helper T (Th2) cell type cytokines (such as interleukin [IL] 4, IL-5, and IL-13) [[6], [7], [8], [9]]. IL-4 promotes the production of IgE, which binds to and activates the surface receptors of mast cells to release a variety of inflammatory factors, leading to bronchial smooth muscle spasm and luminal stenosis [10,11]. IL-5 regulates the growth, differentiation, recruitment and survival of eosinophils, which plays an important role in asthma inflammation and airway remodeling [12]. IL-13 is critical in airway hyperactivity (AHR) and airway remodeling, mucus hypersecretion, and eosinophilic inflammation [13]. Therefore, research related to Th2 type cytokines has become a focus area of allergic asthma research [14,15]. However, in recent years, with the further study of asthma, the role of other immune cells (such as macrophages, inflammatory monocytes, etc.) in allergic asthma has gradually been revealed [16,17].

Recent studies have shown that inflammatory monocytes play a decisive role in the development of ovalbumin (OVA)-induced allergic asthma [16]. Therefore, reducing the number of inflammatory monocytes in the abdominal cavity might be an effective treatment strategy for allergic asthma. The chemokine (C-C motif) ligand 2 (CCL2) is a small cytokine that belongs to the CC chemokine family, which functions in the recruitment of monocytes to inflammation sites [18]. Previous studies have demonstrated that either blocking the CCL2 receptor C-C chemokine receptor type 2 (CCR2) or neutralizing CCL2 could effectively block the recruitment of inflammatory monocytes to inflammatory tissues [19]. Our present study aimed to investigate the role of inflammatory monocytes and CCL2 in mouse model with OVA-induced allergic asthma.

Section snippets

Mouse model of OVA-induced allergic asthma

Mice were initially sensitized by peritoneal administration at day 0 and day 7 with 100 μg of OVA (grade V; Sigma-Aldrich, St. Louis, MO) in 100 μL of sterile saline and adsorbed in 50 μL of Imject alum (Thermo Scientific, Waltham, MA USA). Mice were subsequently challenged by intranasal administration with 100 μg of OVA in 50 μL of sterile saline after anesthetized at day 14, 15, and 16. Mice were euthanized at day 21. Animal study was reviewed and approved by Yuhuangding Hospital.

Collection of BALF

Before the

Mouse model of OVA-induced allergic asthma

To explore the underlying mechanism and efficacious treatment of allergic asthma, we first generated the OVA-induced mouse model (Fig. 1A). Mice were initially sensitized by peritoneal administration with OVA/alum at day 0 and day 7, and subsequently challenged by intranasal administration with OVA protein at day 14, 15, and 16 to induce allergic asthma. In order to verify that the model was successfully built, we collected the bronchoalveolar lavage fluid (BALF) from normal and asthmatic mice

Discussion

Currently, symptoms of allergic asthma can be prevented by avoiding exposure to allergens and irritants and inhaling corticosteroids, but there is no cure for allergic asthma [20]. Therefore, in-depth research to elucidate the pathogenesis of allergic asthma is of great significance for the control and treatment of this disease. In order to explore the underlying mechanism and efficacious treatment, we first generated the OVA-induced mouse model. The OVA-induced mice exhibited an increase of

Conclusion

In conclusion, we successfully constructed a mouse model of allergic asthma induced by OVA and found that the number of inflammatory monocytes in PLF of asthmatic mice was significantly higher than that of normal mice. Down-regulation of CCR2 can significantly reduce the recruitment of inflammatory monocytes into the peritoneal cavity and alleviate the inflammatory response in mice. Blocking CCL2/CCR2 significantly reduced the expression of T2 cytokines and increased the expression of Th1

Declaration of Competing Interest

The authors declare that they have no conflict of interest.

Acknowledgments

None.

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¹: These authors contributed equally to this work.

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Blockade of CCL2/CCR2 signaling pathway prevents inflammatory monocyte recruitment and attenuates OVA-Induced allergic asthma in mice

Highlights

Abstract

Introduction

Section snippets

Mouse model of OVA-induced allergic asthma

Collection of BALF

Mouse model of OVA-induced allergic asthma

Discussion

Conclusion

Declaration of Competing Interest

Acknowledgments

J. Allergy Clin. Immunol. Pract.

Ann. Allergy Asthma Immunol.

Ann. Allergy Asthma Immunol.

J. Allergy Clin. Immunol.

Curr. Opin. Immunol.

J. Autoimmun.

Cytokine

Lancet Oncol.

Biomed. Pharmacother.

Hum. Immunol.

Blood

Summary health statistics for u.s. Adults: national health interview survey, 2011

Vital Health Stat.

Scottish Intercollegiate Guidelines N. British guideline on the management of asthma

Thorax

Epidemiology and economic burden of asthma

Int. Forum Allergy Rhinol.

Allergic vs nonallergic asthma: what makes the difference?

Allergy

Exhaled nitric oxide predicts lung function decline in difficult-to-treat asthma

Eur. Respir. J.

Regulation of IL4 gene expression by T cells and therapeutic perspectives

Nat. Rev. Immunol.