Elsevier

Virus Research

Volume 286, September 2020, 198062
Virus Research

Increased serum levels of galectin-9 in patients with chikungunya fever

https://doi.org/10.1016/j.virusres.2020.198062Get rights and content

Highlights

  • Chikungunya Fever is an arboviral disease that often evolves to a chronic inflammatory musculoskeletal symptoms.

  • Increased levels of galectin-9 have implications in delaying viral clearance, by reducing TCD8+ response.

  • Galectins 9 in involved in the physiopathology of arboviral diseases, no previous information in Chikungunya Fever is available.

  • Galectin-9 is increased in chikungunya chronic disease and its levels are associated with inflammatory symptoms.

  • Galectin-3 showed lower levels in CHIK patients, while other galectins (1,4, and 7) were similar in patients and controls.

Abstract

Chikungunya fever (CHIKF) is an arboviral disease that has caused an epidemic burst of chronic inflammatory joint disease in Latin America in the last few years. Efforts are being spent in understanding the mechanisms by which it may cause such articular damage and in determining possible biomarkers of the disease. Galectins (GAL) are a family of animal lectins with an affinity for beta-galactosides. They have multiple functions including working as receptors in innate immunity and as a control for inflammatory responses in both innate and adaptive immunity. They regulate functions of immune cells, such as lymphocytes and macrophages, which have a main role in the chikungunya inflammatory process. Galectins are also involved in chronification of viral diseases, participate in the immunopathogenesis of chronic joint diseases such as rheumatoid arthritis, and have a role in inflammation in other arboviral diseases, such as dengue. Thus, we intended to determine the serum levels of galectin-1, -3, -4, -7, and -9 in patients with subacute and chronic articular manifestations of CHIKF and to evaluate their associations with clinical manifestations. We evaluated 44 patients with clinical manifestations of CHIKF and serological confirmation with IgM and/or IgG chikungunya virus (CHIKV) antibodies. Forty-nine age- and gender-matched healthy individuals served as controls. Anti−CHIKV IgM and IgG antibodies and galectins serum levels were measured by ELISA. We found higher levels of GAL-9 (patients median 2192 [1500–2631] pg/mL, controls median 46.88 [46.88−46.88] pg/mL, p < 0.0001) and lower levels of GAL-3 (patients median 235.5 [175.5–351.8] pg/mL, controls median 2236.0 [1256.0–2236.0] pg/mL, p < 0.0001) in patients than in controls. There was no statistical difference in levels of GAL-1, -4 and -7 between patients and control groups. There was no difference in GAL-9 serum levels between patients with subacute or chronic symptoms (median 2148 [1500–2722] pg/mL x 2212 [1844–2500] pg/mL, p = 0.3626). A significant association of GAL-9 with joint stiffness, both in its duration and intensity, was found. These results may reflect the participation of GAL-9 in the immunopathogenesis of the inflammatory process in chikungunya fever, as morning stiffness may reflect the systemic inflammatory process.

Introduction

Chikungunya fever (CHIKF) is an arboviral disease caused by the chikungunya virus (CHIKV), and it recently caused an outbreak of continental proportions. The epidemic burst spanned all Latin America, including Brazil, where 700,000 individuals were infected from 2014 to 2019 (Ministério da Saúde, Brasil, 2020). These arboviruses are characterized by acute, intense inflammatory manifestations, evolving with a high symptom chronification rate, with musculoskeletal manifestation predominance, causing critical physical disability and significantly impacting patients’ quality of life.

In the physiopathologic mechanisms of CHIKV disease, the following participate: pro-inflammatory cytokines (IFNs I α and β, IFNγ, IL-4, IL-6, IL-2, TNF-α, TGF-β, IL-12, IL-17A, IL-18, and IL-27), anti-inflammatory cytokines (IL-1Ra, IL-10, and IL-13), chemokines (MCP-1,-2 and -3, MIP-1α and MIP-1β), and growth factors (GM-CSF) (Teng et al., 2015; Cavalcanti et al., 2019). From a cellular point of view, the participation of fibroblastic, neutrophilic, and monocytic-macrophagic lineages T CD4, CD8, NK, and B lymphocytes and osteoclastic activation occur in different degrees of intensity, according to the phase of the disease (Thanapati et al., 2017; Thangamani et al., 2010; Teo et al., 2013; Tanabe et al., 2018).

The chronification process is not completely understood. Autoimmune and auto-inflammatory mechanisms are considered to take part in its development, possibly induced by molecular mimicry to viral antigens. This hypothesis is supported by clinical and physiopathological resemblances between chronic chikungunya and rheumatologic diseases such as rheumatoid arthritis and ankylosing spondylitis (Tanay, 2017; Arroyo-Avila and Vila, 2015).

Galectins (GAL) are of the family of animal origin Lectins, with high affinity for beta-galactosides. They have multiple functions in the regulation of cellular and tissular homeostasis and are capable of recognizing glycidic chains in glycolipids and glycoproteins. They are, for instance, involved in the modulation of cell-cell and cell-matrix interactions (adhesion and signaling) (10). They participate in the process of embryonic development, angiogenesis, tissue regeneration, regulation of cell cycle, adipogenesis, and homeostatic control of tissues. Such diversity of function is dependent on the biochemical conditions of the medium and presence or absence of specific ligands in the tissue microenvironment. The galectin concentration is also relevant for its function, as in high concentration a galectin may bind to an increased number of different ligands, and in low concentration the possible ligands on the cell surface and tissue matrix are more limited (Almkvist and Karlsson, 2002; Barondes et al., 1994; Boscher et al., 2011; Cummings et al., 2015). Galectins play multiple roles in the immunological system. They may function as pathogen- or damage-associated molecular pattern (PAMP/DAMP) receptors, taking part in the antiviral and antimicrobial innate response. Galectins may also exert a regulatory role over innate and adaptive immune responses (Vasta, 2009), including the participation of GAL-9 in Th1/Th17 mediated autoimmunity mechanisms (de la Fuente et al., 2012) and involvement of GAL-1, -3 e -9 in viral infections (Merani et al., 2015).

The participation of galectins in the physiopathological process of rheumatologic inflammatory diseases such as RA (Xie et al., 2018; Pearson et al., 2018; Issa et al., 2017a) and other arboviral diseases such as dengue (Dapat et al., 2017; Liu et al., 2016) drives the interest in evaluating their involvement in the pathogenesis of CHIKF. Beyond that, galectins play an established role in the control of diverse macrophagic and lymphocytic functions (Almkvist and Karlsson, 2002; Sano et al., 2000; Liu et al., 2012; Sano et al., 2003), cells that are intensely involved in the pathogenic process of chikungunya arthropathy (Gardner et al., 2010; Kumar et al., 2012). Being a key regulator of apoptosis, galectins may have a role in the process of chikungunya chronic arthropathy, as experimental evidence points to a deregulated apoptotic process in the genesis of this chronic viral infection (Dhanwani et al., 2012, 2011; Hussain et al., 2016; Sharma et al., 2015).

Section snippets

Methods

We conducted a cross-sectional study to examine the serum levels of galectin-1, -3, -4, -7, and -9 in patients with subacute and chronic articular manifestations of CHIKF and to evaluate their associations with clinical manifestations. The recruited individuals had visited the rheumatology outpatient clinic at the Hospital das Clínicas da Universidade Federal de Pernambuco, from April 2016 to February 2017, with complaints of chronic musculoskeletal symptoms or persistent arthralgia/arthritis.

Clinic-epidemiological data

We evaluated 44 patients over 18 years old with musculoskeletal manifestations of CHIKF in the subacute (3–12 weeks) and chronic phases (>12 weeks), and 49 controls, age- and sex-paired. The clinical and epidemiologic data of patients and controls are exhibited in Table 1.

GAL-9 was detected in most patients (median 2192 [1500–2631] pg/mL) but not in the controls (median 46.88 [46.88−46.88] pg/mL), p < 0.0001. We found decreased serum levels of GAL-3 in patients (median 235.5 [175.5–351.8]

Discussion

We found increased GAL-9 serum levels positively correlated with morning stiffness and associated with greater than 30 min of morning stiffness. Morning stiffness is considered a symptom related to the intensity of joint inflammation and is used as an index sign of disease activity for inflammatory joint diseases such as rheumatoid arthritis (Sierakowski and Cutolo, 2011); therefore, this positive correlation indirectly reflects the liaison of GAL-9 with joint inflammation.

Galectin-9 is

Conclusions

Compared with healthy individuals, we found an increase in GAL-9 levels in chikungunya patients with subacute and chronic musculoskeletal manifestations, with higher levels among chronic phase patients. GAL-3 levels were decreased compared to controls in our analysis. GAL-9 levels correlated with the intensity of morning stiffness and were associated with its duration. As a link between innate and adaptive immunity, and by its relation to the chronicity of viral diseases, galectin-9 may be an

CRediT authorship contribution statement

Nara Gualberto Cavalcanti: Conceptualization, Methodology, Investigation, Methodology, Writing - original draft, Project administration, Software. Kamila Melo Vilar: Investigation, Methodology. Angela Luzia Branco Pinto Duarte: Investigation. Moacyr Jesus Barreto de Melo Rêgo: Resources, Visualization, Validation. Michelly Cristiny Pereira: Resources, Visualization, Writing - review & editing. Ivan da Rocha Pitta: Funding acquisition. Claudia Diniz Lopes Marques: Investigation, Writing - review

Declaration of Competing Interest

The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Acknowledgments

The authors would like to thank the research funding agencies and CNPq, CAPES and FACEPE for the scholarships granted to the researchers participating in the study,

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