Dengue is caused by one of four serotypes of dengue virus (DENV1-4) transmitted by Aedes Aegypti and A. Albopictus mosquitoes, affecting around 400 million people in 128 countries (Bhatt et al., 2013). Occasionally, dengue infection develops into a potentially lethal complication identified as severe dengue, typified by exacerbated systemic inflammation, vascular leakage, fluid accumulation, respiratory distress, severe bleeding, and/or organ impairment (WHO, 2009). No antiviral drug for dengue treatment is available, and the use of Dengvaxia (CYD-TDV), the first dengue vaccine approved by the US Food and Drug Administration, has its limitations, such as the increased risk for development of severe dengue in the immune populations (Huang et al., 2021; WHO, 2018; The Lancet Infectious Diseases, 2018). Thus, the combination of these factors points to dengue as a major unmet clinical problem in countries affected by this disease (Shepard et al., 2016; Stanaway et al., 2016).

The pathogenesis of severe dengue results from exacerbated host innate and adaptative immune responses to DENV. Among the target cells for DENV in humans, mast cells (MCs) lining blood vessels undergo dramatic cellular activation in response to DENV, despite their high resistance to infection (; St John et al., 2013bSt John et al., 2011). As essential regulators of vascular integrity, MC activation evoked by DENV triggers the production of inflammatory cytokines (cytokine storm) and vascular leakage, which ultimately result in hypovolemic shock in severe dengue (St John et al., 2013b; Syenina et al., 2015St John et al., 2011;). This is substantiated by a positive correlation between circulating chymase levels and disease severity recently observed in paediatric and adult patients (Rathore et al., 2020; Tissera et al., 2017). In line with the suggested role of uncontrolled immune responses in the pathogenesis of DENV infection, pharmacological suppression of inflammation (Fu et al., 2014; Marques et al., 2015; Souza et al., 2009), MC stabilisation (St John et al., 2013bMorrison et al., 2017), and inhibition of MC-derived protease (Rathore et al., 2019) have shown to be beneficial in experimental dengue. These studies provide proof-of-concept that host-directed therapies targeting excessive or misplaced inflammation may be a viable approach in treating severe dengue infection.

Pro-resolving mediators are cell signalling molecules synthesised in a strict temporal and spatial fashion to regulate the host response and prevent the excessive acute inflammatory reaction that damages the host (Sugimoto et al., 2019). The discovery of this active phase of inflammation has led to a new awareness of how a disease can emerge, including the concept that dysregulation or ‘failure’ in pro-resolving mechanisms might be involved in the pathogenesis of several chronic inflammatory disorders (Eke Gungor et al., 2014; Fredman et al., 2016; Murri et al., 2013; Schett and Neurath, 2018; Sena et al., 2013; Thul et al., 2017; Vong et al., 2012). The pro-resolving protein Annexin A1 (AnxA1) and its cognate receptor formyl peptide receptor 2 (FPR2), also known as FPR2/lipoxin A4 receptor (FPR2/ALX), bear anti-inflammatory properties in sterile settings (Fredman et al., 2015; Galvão et al., 2017; Gimenes et al., 2015; Kusters et al., 2015; Leoni et al., 2015; Locatelli et al., 2014), and exert a degree of protection in infectious settings, such as experimental tuberculosis (Tzelepis et al., 2015; Vanessa et al., 2015), sepsis (Damazo et al., 2005; Gobbetti et al., 2014) Spite et al., 2009, pneumococcal pneumonia (Machado et al., 2020; Tavares et al., 2016), and influenza (Schloer et al., 2019). AnxA1 has been recently described to act as an endogenous modulator of MC degranulation in response to IgE/anti-IgE or compound 48/80 (Parisi et al., 2019; Sinniah et al., 2019; Sinniah et al., 2016; Yazid et al., 2013). Since AnxA1 is well known to counter regulate overexuberant pro-inflammatory events and MC activation, we have hypothesised that an imbalance between this anti-inflammatory/pro-resolving mediator and pro-inflammatory molecules could be operating during dengue infection. Whilst studies on bacterial infection consistently revealed the ability of specialised pro-resolving mediators and AnxA1 (Chiang et al., 2012; Decker et al., 2021; Gobbetti et al., 2014; Machado et al., 2020; Sekheri et al., 2020; Tzelepis et al., 2015; Vanessa et al., 2015) to facilitate innate immune responses against the pathogen whilst reducing the harmful effects of inflammation, the translational potential of these results to viral infection was less clear.

In the present work, we have analysed the role of the pro-resolving AnxA1-FPR2/ALX pathway as a regulator of excessive inflammation observed in patients with the most severe forms of dengue infection. Our results suggest that failure to trigger this molecular pathway may contribute to disease severity in dengue infection and support the AnxA1-FPR2/ALX pathway as a potential target for host-directed therapy in human dengue disease.

We present evidence that an inadequate engagement of the resolution circuit AnxA1-FPR2/ALX may contribute to dengue infection’s pathogenesis with particular relevance for the cohort of patients affected by the most severe forms of the disease. By exploring resolution biology as a novel approach in dengue disease, both with respect to etiopathogenesis and pharmacological opportunity, we have identified that: (i) AnxA1 is downregulated in the plasma of dengue patients in comparison to healthy individuals and in DENV-infected mice in comparison to non-infected animals; (ii) depletion of the AnxA1-FPR2/ALX pathway aggravates clinical signs and enhances MC activation associated to DENV infection, indicating a nonredundant role for this resolution pathway in the pathogenesis of dengue disease; (iii) pharmacological treatment of mice with an FPR2/ALX agonistic peptide produced beneficial effects during DENV infection; (iv) AnxA1 mimetic peptide has direct inhibitory effects on MC degranulation induced by DENV while (v) it does not seem to control viral load and virus dissemination significantly.

In recent years, a new paradigm shift has emerged in our understanding of the pathogenesis of inflammatory diseases, which results from persistent and exacerbated pro-inflammatory signals and dysregulation or ‘failure’ in resolving mechanisms (Perucci et al., 2017) Eke Gungor et al., 2014; Fredman et al., 2016; Murri et al., 2013; Schett and Neurath, 2018; Sena et al., 2013; Thul et al., 2017; Vong et al., 2012. The severity and lethality of several infectious diseases, like dengue and the flu, frequently arise from an excessive host response, characterised by an uncontrolled release of pro-inflammatory cytokines leading to over-exuberant immune activation (Costa et al., 2013; St John et al., 2013a). Inflammation is physiologically balanced by resolution circuits, such as those centred on AnxA1 and lipid mediators (e.g. lipoxins, resolvins, protectins and maresins) that drive termination of inflammatory response yet helping the host to deal with the infective agent (Basil and Levy, 2016). In line with this, a growing body of evidence indicates that pro-resolving mediators are regulated during infection, contributing to the control and resolution of experimental and human infectious diseases (Abdulnour et al., 2016; Basil and Levy, 2016; Chiang et al., 2012; Frediani et al., 2014; Oliveira et al., 2017; Shirey et al., 2014). On the other hand, dysregulation in the production and/or action of pro-resolving mediators might contribute to the pathogenesis of sterile (Eke Gungor et al., 2014; Fredman et al., 2016; Murri et al., 2013; Sena et al., 2013; Thul et al., 2017; Vong et al., 2012) and infectious (Cilloniz et al., 2010; Colas et al., 2019; Molás et al., 2020; Morita et al., 2013) diseases, including atherosclerosis, inflammatory bowel diseases and tuberculous meningitis. Modulation of AnxA1 in the context of viral infections has been less investigated, and clinical data are scarce (Arora et al., 2016; Molás et al., 2020). Since recent evidence suggests a defective engagement of pro-resolving pathways during self-resolving infections (Cilloniz et al., 2010; Colas et al., 2019; Molás et al., 2020; Morita et al., 2013), we queried whether such alterations could also be occurring in dengue disease. Focus was given to the pro-resolving protein AnxA1 and its cognate receptor FPR2/ALX, a pathway that has been shown to exert a degree of protection in experimental tuberculosis (Tzelepis et al., 2015; Vanessa et al., 2015), sepsis (Damazo et al., 2005; Gobbetti et al., 2014), pneumococcal pneumonia (Machado et al., 2020; Tavares et al., 2016), and influenza (Schloer et al., 2019). Our data indicate that in addition to the already described early induction of an inflammatory response that may be harmful instead of protective, DENV infection is also characterised by sustained downregulation of molecular components of the AnxA1 pathway. The drop in circulating AnxA1 below basal levels observed in dengue patients in this study, especially in severe dengue, suggests that the pathogenesis and severity of the disease might be associated with a failure to engage mechanisms involved in endogenous anti-inflammatory signals and its resolution, such as those centred on AnxA1. Furthermore, we have identified a protective role for the AnxA1-FPR2/ALX pathway in DENV infection, as animals presented heightened signs of disease in the absence of either ligand or receptor, compared to WT mice. In line with integrated pro-resolving properties of resolution circuits, AnxA1 and FPR2 KO mice displayed diverse uncontrolled responses leading to increased disease severity. Markers of MC degranulation and systemic inflammation were elevated in transgenic mice compared to WT counterparts. Similarly, these genetically manipulated animals presented enhanced haematological alterations in our model of experimental dengue.

Infections are currently treated by drugs that target pathogens or inhibit their growth. In some infectious diseases, blocking inflammation pathways may be beneficial. While this approach might be successful in some infections, excessive inhibition of the immune response can also be associated with immunosuppression and increased mortality, as observed in septic patients (Carlet et al., 2020). Resolution pharmacology has been proposed as an alternative to balance the host response without hampering its ability to deal with the infection. Indeed, pro-resolving receptor agonists’ exogenous administration has proven benefits in experimental infectious settings, including bacterial pneumonia (Abdulnour et al., 2016; Machado et al., 2020) and influenza (Morita et al., 2013; Ramon et al., 2014; Schloer et al., 2019). In the present work, we provide proof-of-principle that a pro-resolving tool, the Ac_2-26 peptide, can ameliorate clinical disease and reduce circulating inflammatory mediators in an FPR2/ALX-dependent manner. The AnxA1 peptide was effective even when administered from day 2 after the infection onset.

In our experimental models, viral load was seemingly unaffected by Ac_2-26 treatment, supporting a protective effect independent of viral infectivity. Despite the limitations of modelling dengue disease in immunocompetent animals, infection of AnxA1 KO and FPR2 KO animals (and their respective littermates BALB/c and C57BL/6 mice) with high inoculum of DENV-2 was applied to investigate the role of endogenous AnxA1/FPR2 in dengue disease. Although viremia is not detectable in immunocompetent mice, we noticed transient viral RNA detection in the spleen starting 24 hr after infection, peaking at 48 hr or 72 hr, followed by disappearance at 96 hr, in line with previous studies using the same model (St John et al., 2013b; Syenina et al., 2015). Neither genetic deletion of AnxA1 or treatment with Ac_2-26 affected this profile. To complement our studies with immunocompetent animals mice, we performed experiments in A129 mice since they are known to be very susceptible to flaviviruses infections (de Freitas et al., 2019; Del Sarto et al., 2020) and develop a more severe disease manifestation that emulates several features of severe dengue in humans (Zandi et al., 2019). The later is considered a gold standard viremia model to test potential antiviral therapies (Meyts and Casanova, 2021; Williams et al., 2009). Using this model, we aimed to evaluate if the treatment with peptide Ac_2-26 has any impact on viral loads as well as in disease and inflammatory parameters. Of note, we were able to address the quantification of the viral loads by the plaque assay technique, the gold standard methodology to evaluate viable viral particles in vivo. Therefore, in our study we show, by two different techniques, that is, quantification of viral RNA (immunocompetent mice) and quantification of viable particles (A129 mice), and in two different models (high and low DENV inoculums) that neither absence of AnxA1 nor treatment with peptide Ac_2-26 interferes with the host ability to deal with the infection, despite its beneficial pro-resolving effects.

DENV infection initiates pro-inflammatory responses aiming to control virus spread that ultimately contributes to the immunopathology of dengue. For instance, MC activation in response to DENV plays an essential role in DENV-induced vascular pathology, particularly concerning the plasma leakage that causes hypovolemic shock in severe dengue (St John et al., 2013b; Syenina et al., 2015). This is supported by a correlation between circulating chymase levels and disease severity in humans (Rathore et al., 2020; Tissera et al., 2017). It has been recently described that AnxA1 acts as an endogenous modulator of MC degranulation in response to IgE/anti-IgE or compound 48/80, suggesting that this pro-resolving axis act as a brake in MC degranulation (Sinniah et al., 2016; Yazid et al., 2013). The present study confirmed that dengue disease is associated with increased plasma levels of MCPT-1 and identified enhanced secretion in animals lacking AnxA1 or FPR2/ALX. Herein, we have identified the ability of Ac_2-26 to reduce DENV-induced MC degranulation dose-dependently, a mechanism that might underpin the reduced MCPT-1 secretion and vascular dysfunction observed in AnxA1 peptide-treated animals. Together, in vivo and in vitro evidence suggest that Ac_2-26, at least in part, acts by attenuating MC degranulation evoked by DENV, protecting the host against vascular dysfunction associated with the disease. This mode of action points to this pathway as a relevant potential target for DENV infection treatment, as MCs are resistant to infection but play a key role in dengue pathogenesis (; Rathore et al., 2019; Syenina et al., 2015; Tissera et al., 2017).

Although no therapy is available for dengue disease beyond supportive care, recent studies have pointed towards a benefit of using corticoids in severe dengue with no significant adverse consequences or promotion of virus replication (Bandara and Herath, 2018; Bandara and Herath, 2020). Amongst the several genomic and non-genomic actions of glucocorticoids (GC), promotion of AnxA1 has shown to be relevant to the benefits afforded by steroids (De Caterina et al., 1993; Solito et al., 2003), as suggested by the loss of the therapeutic effect of GC on AnxA1 depleted animals (Patel et al., 2012; Perretti and D’Acquisto, 2009; Vago et al., 2012). Our study reveals a central role of the axis AnxA1-FPR2/ALX in the pathogenesis of dengue disease. We suggest that this critical anti-inflammatory/pro-resolving pathway is downregulated during dengue, contributing to unbalanced inflammation and the cytokine storm characteristic of the disease. Therefore, there is scope to hypothesise that the benefit of glucocorticoid in dengue disease could be partially attributed to promoting the glucocorticoid inducible Annexin A1-FPR2/ALX pathway. Further investigations should confirm AnxA1 as an underlying mechanism of GCs in dengue disease. The effectiveness of the treatment with AnxA1 peptide in our study suggests that GC therapy could be refined using a more targeted approach, particularly in infectious diseases where there is an underlying imbalance in the AnxA1 pathway. This opens an opportunity to explore small molecules targeting FPR2/ALX receptors, avoiding the spanning adverse consequences of glucocorticoids whilst targeting the defect in inflammation resolution operating in the disease settings. Further investigation on GC and GC-inducible AnxA1 is particularly relevant in the context of dengue disease, where the use of non-steroidal anti-inflammatory agents in dengue is discouraged, as they can increase the risk of bleeding (WHO, 2009).

Our results indicate that altered levels of the pro-resolving mediator AnxA1 are of pathological relevance in dengue disease. We show that inadequate engagement of resolution circuits contributes to the excessive inflammation observed in severe DENV infection. In addition, we provide evidence for the benefits of pharmacological therapy directed to modulating host immune responses in the absence of a direct antiviral effect. These findings point to a direction for future research on applying FPR2/ALX agonists as a therapeutic target for dengue disease. Since pathogenic immune responses are not exclusive to dengue but underlie the severity of several other viral diseases, including severe community-acquired pneumonia caused by viruses (D’Elia et al., 2013; Perrone et al., 2008) and COVID-19 (Jose and Manuel, 2020), our findings could be translated into different infectious settings, whereby targeting the AnxA1 pathway, with or without combination with antiviral drugs, holds promising therapeutic potential.

All data has been included in the manuscript, and source data files have been provided for Figures 1-6.

1. 1. Abdulnour RE
   2. Sham HP
   3. Douda DN
   4. Colas RA
   5. Dalli J
   6. Bai Y
   7. Ai X
   8. Serhan CN
   9. Levy BD

   (2016) Aspirin-triggered resolvin D1 is produced during self-resolving gram-negative bacterial pneumonia and regulates host immune responses for the resolution of lung inflammation

   Mucosal Immunology 9:1278–1287.

   https://doi.org/10.1038/mi.2015.129

   • PubMed
   • Google Scholar
2. 1. Andrade MV
   2. Iwaki S
   3. Ropert C
   4. Gazzinelli RT
   5. Cunha-Melo JR
   6. Beaven MA

   (2011) Amplification of cytokine production through synergistic activation of NFAT and AP-1 following stimulation of mast cells with antigen and IL-33

   European Journal of Immunology 41:760–772.

   https://doi.org/10.1002/eji.201040718

   • PubMed
   • Google Scholar
3. 1. Arora S
   2. Lim W
   3. Bist P
   4. Perumalsamy R
   5. Lukman HM
   6. Li F
   7. Welker LB
   8. Yan B
   9. Sethi G
   10. Tambyah PA
   11. Fairhurst A-M
   12. Alonso S
   13. Lim LHK

   (2016) Influenza A virus enhances its propagation through the modulation of Annexin-A1 dependent endosomal trafficking and apoptosis

   Cell Death and Differentiation 23:1243–1256.

   https://doi.org/10.1038/cdd.2016.19

   • PubMed
   • Google Scholar
4. 1. Bandara SMR
   2. Herath H

   (2018) Effectiveness of corticosteroid in the treatment of dengue - A systemic review

   Heliyon 4:e00816.

   https://doi.org/10.1016/j.heliyon.2018.e00816

   • PubMed
   • Google Scholar
5. 1. Bandara SMR
   2. Herath H

   (2020) Corticosteroid actions on dengue immune pathology; A review article

   Clinical Epidemiology and Global Health 8:486–494.

   https://doi.org/10.1016/j.cegh.2019.11.001

   • Google Scholar
6. 1. Basil MC
   2. Levy BD

   (2016) Specialized pro-resolving mediators: endogenous regulators of infection and inflammation

   Nature Reviews. Immunology 16:51–67.

   https://doi.org/10.1038/nri.2015.4

   • PubMed
   • Google Scholar
7. 1. Bhatt S
   2. Gething PW
   3. Brady OJ
   4. Messina JP
   5. Farlow AW
   6. Moyes CL
   7. Drake JM
   8. Brownstein JS
   9. Hoen AG
   10. Sankoh O
   11. Myers MF
   12. George DB
   13. Jaenisch T
   14. Wint GRW
   15. Simmons CP
   16. Scott TW
   17. Farrar JJ
   18. Hay SI

   (2013) The global distribution and burden of dengue

   Nature 496:504–507.

   https://doi.org/10.1038/nature12060

   • PubMed
   • Google Scholar
8. 1. Carlet J
   2. Payen D
   3. Opal SM

   (2020) Steroids for sepsis and ARDS: this eternal controversy remains with COVID-19

   Lancet (London, England) 396:e61–e62.

   https://doi.org/10.1016/S0140-6736(20)32132-2

   • PubMed
   • Google Scholar
9. 1. Carmo LAS
   2. Bonjour K
   3. Ueki S
   4. Neves JS
   5. Liu L
   6. Spencer LA
   7. Dvorak AM
   8. Weller PF
   9. Melo RCN

   (2016) CD63 is tightly associated with intracellular, secretory events chaperoning piecemeal degranulation and compound exocytosis in human eosinophils

   Journal of Leukocyte Biology 100:391–401.

   https://doi.org/10.1189/jlb.3A1015-480R

   • PubMed
   • Google Scholar
10. 1. Chen HC
    2. Hofman FM
    3. Kung JT
    4. Lin YD
    5. Wu-Hsieh BA

    (2007) Both virus and tumor necrosis factor alpha are critical for endothelium damage in a mouse model of dengue virus-induced hemorrhage

    Journal of Virology 81:5518–5526.

    https://doi.org/10.1128/JVI.02575-06

    • PubMed
    • Google Scholar
11. 1. Chiang N
    2. Fredman G
    3. Bäckhed F
    4. Oh SF
    5. Vickery T
    6. Schmidt BA
    7. Serhan CN

    (2012) Infection regulates pro-resolving mediators that lower antibiotic requirements

    Nature 484:524–528.

    https://doi.org/10.1038/nature11042

    • PubMed
    • Google Scholar
12. 1. Cilloniz C
    2. Pantin-Jackwood MJ
    3. Ni C
    4. Goodman AG
    5. Peng X
    6. Proll SC
    7. Carter VS
    8. Rosenzweig ER
    9. Szretter KJ
    10. Katz JM
    11. Korth MJ
    12. Swayne DE
    13. Tumpey TM
    14. Katze MG

    (2010) Lethal dissemination of H5N1 influenza virus is associated with dysregulation of inflammation and lipoxin signaling in a mouse model of infection

    Journal of Virology 84:7613–7624.

    https://doi.org/10.1128/JVI.00553-10

    • PubMed
    • Google Scholar
13. 1. Colas RA
    2. Nhat LTH
    3. Thuong NTT
    4. Gómez EA
    5. Ly L
    6. Thanh HH
    7. Mai NTH
    8. Phu NH
    9. Thwaites GE
    10. Dalli J

    (2019) Proresolving mediator profiles in cerebrospinal fluid are linked with disease severity and outcome in adults with tuberculous meningitis

    FASEB Journal 33:13028–13039.

    https://doi.org/10.1096/fj.201901590R

    • PubMed
    • Google Scholar
14. 1. Combs JW

    (1966) Maturation of rat mast cells. An electron microscope study

    The Journal of Cell Biology 31:563–575.

    https://doi.org/10.1083/jcb.31.3.563

    • PubMed
    • Google Scholar
15. 1. Combs JW

    (1971) An electron microscope study of mouse mast cells arising in vivo and in vitro

    The Journal of Cell Biology 48:676–684.

    https://doi.org/10.1083/jcb.48.3.676

    • PubMed
    • Google Scholar
16. 1. Costa VV
    2. Fagundes CT
    3. Valadão DF
    4. Cisalpino D
    5. Dias ACF
    6. Silveira KD
    7. Kangussu LM
    8. Ávila TV
    9. Bonfim MRQ
    10. Bonaventura D
    11. Silva TA
    12. Sousa LP
    13. Rachid MA
    14. Vieira LQ
    15. Menezes GB
    16. de Paula AM
    17. Atrasheuskaya A
    18. Ignatyev G
    19. Teixeira MM
    20. Souza DG

    (2012) A model of DENV-3 infection that recapitulates severe disease and highlights the importance of IFN-gamma in host resistance to infection

    PLOS Neglected Tropical Diseases 6:e1663.

    https://doi.org/10.1371/journal.pntd.0001663

    • PubMed
    • Google Scholar
17. 1. Costa VV
    2. Fagundes CT
    3. Souza DG
    4. Teixeira MM

    (2013) Inflammatory and innate immune responses in dengue infection: protection versus disease induction

    The American Journal of Pathology 182:1950–1961.

    https://doi.org/10.1016/j.ajpath.2013.02.027

    • PubMed
    • Google Scholar
18. 1. Costa VV
    2. Fagundes CT
    3. Valadão DF
    4. Ávila TV
    5. Cisalpino D
    6. Rocha RF
    7. Ribeiro LS
    8. Ascenção FR
    9. Kangussu LM
    10. Celso MQ
    11. Astigarraga RG
    12. Gouveia FL
    13. Silva TA
    14. Bonaventura D
    15. Sampaio D
    16. Leite ACL
    17. Teixeira MM
    18. Souza DG

    (2014) Subversion of early innate antiviral responses during antibody-dependent enhancement of Dengue virus infection induces severe disease in immunocompetent mice

    Medical Microbiology and Immunology 203:231–250.

    https://doi.org/10.1007/s00430-014-0334-5

    • PubMed
    • Google Scholar
19. 1. Damazo AS
    2. Yona S
    3. D’Acquisto F
    4. Flower RJ
    5. Oliani SM
    6. Perretti M

    (2005) Critical Protective Role for Annexin 1 Gene Expression in the Endotoxemic Murine Microcirculation

    The American Journal of Pathology 166:1607–1617.

    https://doi.org/10.1016/S0002-9440(10)62471-6

    • PubMed
    • Google Scholar
20. 1. Damazo AS
    2. Yona S
    3. Flower RJ
    4. Perretti M
    5. Oliani SM

    (2006) Spatial and temporal profiles for anti-inflammatory gene expression in leukocytes during a resolving model of peritonitis

    Journal of Immunology (Baltimore, Md 176:4410–4418.

    https://doi.org/10.4049/jimmunol.176.7.4410

    • PubMed
    • Google Scholar
21. 1. De Caterina R
    2. Sicari R
    3. Giannessi D
    4. Paggiaro PL
    5. Paoletti P
    6. Lazzerini G
    7. Bernini W
    8. Solito E
    9. Parente L

    (1993) Macrophage-specific eicosanoid synthesis inhibition and lipocortin-1 induction by glucocorticoids

    Journal of Applied Physiology (Bethesda, Md 75:2368–2375.

    https://doi.org/10.1152/jappl.1993.75.6.2368

    • PubMed
    • Google Scholar
22. 1. de Freitas CS
    2. Higa LM
    3. Sacramento CQ
    4. Ferreira AC
    5. Reis PA
    6. Delvecchio R
    7. Monteiro FL
    8. Barbosa-Lima G
    9. James Westgarth H
    10. Vieira YR
    11. Mattos M
    12. Rocha N
    13. Hoelz LVB
    14. Leme RPP
    15. Bastos MM
    16. Rodrigues GOL
    17. Lopes CEM
    18. Queiroz-Junior CM
    19. Lima CX
    20. Costa VV
    21. Teixeira MM
    22. Bozza FA
    23. Bozza PT
    24. Boechat N
    25. Tanuri A
    26. Souza TML

    (2019) Yellow fever virus is susceptible to sofosbuvir both in vitro and in vivo

    PLOS Neglected Tropical Diseases 13:e0007072.

    https://doi.org/10.1371/journal.pntd.0007072

    • PubMed
    • Google Scholar
23. 1. Decker C
    2. Sadhu S
    3. Fredman G

    (2021) Pro-Resolving Ligands Orchestrate Phagocytosis

    Frontiers in Immunology 12:660865.

    https://doi.org/10.3389/fimmu.2021.660865

    • PubMed
    • Google Scholar
24. 1. Del Sarto JL
    2. Rocha R
    3. Bassit L
    4. Olmo IG
    5. Valiate B
    6. Queiroz-Junior CM
    7. Pedrosa C
    8. Ribeiro FM
    9. Guimarães MZ
    10. Rehen S
    11. Amblard F
    12. Zhou L
    13. Cox BD
    14. Gavegnano C
    15. Costa VV
    16. Schinazi RF
    17. Teixeira MM

    (2020) 7-Deaza-7-Fluoro-2'-C-Methyladenosine Inhibits Zika virus Infection and Viral-induced Neuroinflammation

    Antiviral Research 180:104855.

    https://doi.org/10.1016/j.antiviral.2020.104855

    • PubMed
    • Google Scholar
25. 1. Dufton N
    2. Hannon R
    3. Brancaleone V
    4. Dalli J
    5. Patel HB
    6. Gray M
    7. D’Acquisto F
    8. Buckingham JC
    9. Perretti M
    10. Flower RJ

    (2010) Anti-inflammatory role of the murine formyl-peptide receptor 2: ligand-specific effects on leukocyte responses and experimental inflammation

    Journal of Immunology (Baltimore, Md 184:2611–2619.

    https://doi.org/10.4049/jimmunol.0903526

    • PubMed
    • Google Scholar
26. 1. Dufton N
    2. Hannon R
    3. Brancaleone V
    4. Dalli J
    5. Patel HB
    6. Gray M
    7. D’Acquisto F
    8. Buckingham JC
    9. Perretti M
    10. Flower RJ

    (2011) Corrections: Anti-Inflammatory Role of the Murine Formyl-Peptide Receptor 2: Ligand-Specific Effects on Leukocyte Responses and Experimental Inflammation

    The Journal of Immunology 186:2684–2685.

    https://doi.org/10.4049/jimmunol.1090139

    • Google Scholar
27. 1. Dvorak AM
    2. Nabel G
    3. Pyne K
    4. Cantor H
    5. Dvorak HF
    6. Galli SJ

    (1982) Ultrastructural identification of the mouse basophil

    Blood 59:1279–1285.

    https://doi.org/10.1182/blood.V59.6.1279.1279

    • PubMed
    • Google Scholar
28. 1. D’Elia RV
    2. Harrison K
    3. Oyston PC
    4. Lukaszewski RA
    5. Clark GC

    (2013) Targeting the "cytokine storm" for therapeutic benefit

    Clinical and Vaccine Immunology 20:319–327.

    https://doi.org/10.1128/CVI.00636-12

    • PubMed
    • Google Scholar
29. 1. Eke Gungor H
    2. Tahan F
    3. Gokahmetoglu S
    4. Saraymen B

    (2014) Decreased levels of lipoxin A4 and annexin A1 in wheezy infants

    International Archives of Allergy and Immunology 163:193–197.

    https://doi.org/10.1159/000358490

    • PubMed
    • Google Scholar
30. 1. Frediani JK
    2. Jones DP
    3. Tukvadze N
    4. Uppal K
    5. Sanikidze E
    6. Kipiani M
    7. Tran VT
    8. Hebbar G
    9. Walker DI
    10. Kempker RR
    11. Kurani SS
    12. Colas RA
    13. Dalli J
    14. Tangpricha V
    15. Serhan CN
    16. Blumberg HM
    17. Ziegler TR

    (2014) Plasma metabolomics in human pulmonary tuberculosis disease: a pilot study

    PLOS ONE 9:e108854.

    https://doi.org/10.1371/journal.pone.0108854

    • PubMed
    • Google Scholar
31. 1. Fredman G
    2. Kamaly N
    3. Spolitu S
    4. Milton J
    5. Ghorpade D
    6. Chiasson R
    7. Kuriakose G
    8. Perretti M
    9. Farokzhad O
    10. Tabas I

    (2015) Targeted nanoparticles containing the proresolving peptide Ac2-26 protect against advanced atherosclerosis in hypercholesterolemic mice

    Science Translational Medicine 7:275ra220.

    https://doi.org/10.1126/scitranslmed.aaa1065

    • PubMed
    • Google Scholar
32. 1. Fredman G
    2. Hellmann J
    3. Proto JD
    4. Kuriakose G
    5. Colas RA
    6. Dorweiler B
    7. Connolly ES
    8. Solomon R
    9. Jones DM
    10. Heyer EJ
    11. Spite M
    12. Tabas I

    (2016) An imbalance between specialized pro-resolving lipid mediators and pro-inflammatory leukotrienes promotes instability of atherosclerotic plaques

    Nature Communications 7:12859.

    https://doi.org/10.1038/ncomms12859

    • PubMed
    • Google Scholar
33. 1. Fu Y
    2. Yip A
    3. Seah PG
    4. Blasco F
    5. Shi P-Y
    6. Hervé M

    (2014) Modulation of inflammation and pathology during dengue virus infection by p38 MAPK inhibitor SB203580

    Antiviral Research 110:151–157.

    https://doi.org/10.1016/j.antiviral.2014.08.004

    • PubMed
    • Google Scholar
34. 1. Galvão I
    2. Vago JP
    3. Barroso LC
    4. Tavares LP
    5. Queiroz-Junior CM
    6. Costa VV
    7. Carneiro FS
    8. Ferreira TP
    9. Silva PMR
    10. Amaral FA
    11. Sousa LP
    12. Teixeira MM

    (2017) Annexin A1 promotes timely resolution of inflammation in murine gout

    European Journal of Immunology 47:585–596.

    https://doi.org/10.1002/eji.201646551

    • PubMed
    • Google Scholar
35. 1. Gimenes AD
    2. Andrade TRM
    3. Mello CB
    4. Ramos L
    5. Gil CD
    6. Oliani SM

    (2015) Beneficial effect of annexin A1 in a model of experimental allergic conjunctivitis

    Experimental Eye Research 134:24–32.

    https://doi.org/10.1016/j.exer.2015.03.013

    • PubMed
    • Google Scholar
36. 1. Gobbetti T
    2. Coldewey SM
    3. Chen J
    4. McArthur S
    5. le Faouder P
    6. Cenac N
    7. Flower RJ
    8. Thiemermann C
    9. Perretti M

    (2014) Nonredundant protective properties of FPR2/ALX in polymicrobial murine sepsis

    PNAS 111:18685–18690.

    https://doi.org/10.1073/pnas.1410938111

    • PubMed
    • Google Scholar
37. 1. Gonçalves Pereira MH
    2. Figueiredo MM
    3. Queiroz CP
    4. Magalhães TVB
    5. Mafra A
    6. Diniz LMO
    7. da Costa ÚL
    8. Gollob KJ
    9. Antonelli LR
    10. Santiago H

    (2020) T cells producing multiple combinations of IFNgamma, TNF and IL10 are associated with mild forms of dengue infection

    Immunology 160:90–102.

    https://doi.org/10.1111/imm.13185

    • PubMed
    • Google Scholar
38. 1. Hannon R
    2. Croxtall JD
    3. Getting SJ
    4. Roviezzo F
    5. Yona S
    6. Paul-Clark MJ
    7. Gavins FNE
    8. Perretti M
    9. Morris JF
    10. Buckingham JC
    11. Flower RJ

    (2003) Aberrant inflammation and resistance to glucocorticoids in annexin 1-/- mouse

    FASEB Journal 17:253–255.

    https://doi.org/10.1096/fj.02-0239fje

    • PubMed
    • Google Scholar
39. 1. Huang CH
    2. Tsai YT
    3. Wang SF
    4. Wang WH
    5. Chen YH

    (2021) Dengue vaccine: an update

    Expert Review of Anti-Infective Therapy 19:1495–1502.

    https://doi.org/10.1080/14787210.2021.1949983

    • PubMed
    • Google Scholar
40. 1. Hue KDT
    2. Tuan TV
    3. Thi HTN
    4. Bich CTN
    5. Anh HHL
    6. Wills BA
    7. Simmons CP

    (2011) Validation of an internally controlled one-step real-time multiplex RT-PCR assay for the detection and quantitation of dengue virus RNA in plasma

    Journal of Virological Methods 177:168–173.

    https://doi.org/10.1016/j.jviromet.2011.08.002

    • PubMed
    • Google Scholar
41. 1. Jose RJ
    2. Manuel A

    (2020) COVID-19 cytokine storm: the interplay between inflammation and coagulation

    The Lancet. Respiratory Medicine 8:e46–e47.

    https://doi.org/10.1016/S2213-2600(20)30216-2

    • PubMed
    • Google Scholar
42. 1. Kusters DH
    2. Chatrou ML
    3. Willems BA
    4. De Saint-Hubert M
    5. Bauwens M
    6. Vorst E
    7. Reutelingsperger CP

    (2015) Pharmacological Treatment with Annexin A1 Reduces Atherosclerotic Plaque Burden in LDLR-/- Mice on Western Type Diet

    PLOS ONE 10:e0130484.

    https://doi.org/10.1371/journal.pone.0130484

    • Google Scholar
43. 1. Lam JH
    2. Chua YL
    3. Lee PX
    4. Martínez Gómez JM
    5. Ooi EE
    6. Alonso S

    (2017) Dengue vaccine-induced CD8+ T cell immunity confers protection in the context of enhancing, interfering maternal antibodies

    JCI Insight 2:24.

    https://doi.org/10.1172/jci.insight.94500

    • PubMed
    • Google Scholar
44. 1. Leoni G
    2. Neumann P-A
    3. Kamaly N
    4. Quiros M
    5. Nishio H
    6. Jones HR
    7. Sumagin R
    8. Hilgarth RS
    9. Alam A
    10. Fredman G
    11. Argyris I
    12. Rijcken E
    13. Kusters D
    14. Reutelingsperger C
    15. Perretti M
    16. Parkos CA
    17. Farokhzad OC
    18. Neish AS
    19. Nusrat A

    (2015) Annexin A1-containing extracellular vesicles and polymeric nanoparticles promote epithelial wound repair

    The Journal of Clinical Investigation 125:1215–1227.

    https://doi.org/10.1172/JCI76693

    • PubMed
    • Google Scholar
45. 1. Locatelli I
    2. Sutti S
    3. Jindal A
    4. Vacchiano M
    5. Bozzola C
    6. Reutelingsperger C
    7. Kusters D
    8. Bena S
    9. Parola M
    10. Paternostro C
    11. Bugianesi E
    12. McArthur S
    13. Albano E
    14. Perretti M

    (2014) Endogenous annexin A1 is a novel protective determinant in nonalcoholic steatohepatitis in mice

    Hepatology (Baltimore, Md.) 60:531–544.

    https://doi.org/10.1002/hep.27141

    • PubMed
    • Google Scholar
46. 1. Machado MG
    2. Tavares LP
    3. Souza GVS
    4. Queiroz-Junior CM
    5. Ascenção FR
    6. Lopes ME
    7. Garcia CC
    8. Menezes GB
    9. Perretti M
    10. Russo RC
    11. Teixeira MM
    12. Sousa LP

    (2020) The Annexin A1/FPR2 pathway controls the inflammatory response and bacterial dissemination in experimental pneumococcal pneumonia

    FASEB Journal 34:2749–2764.

    https://doi.org/10.1096/fj.201902172R

    • PubMed
    • Google Scholar
47. 1. Marques RE
    2. Guabiraba R
    3. Del Sarto JL
    4. Rocha RF
    5. Queiroz AL
    6. Cisalpino D
    7. Marques PE
    8. Pacca CC
    9. Fagundes CT
    10. Menezes GB
    11. Nogueira ML
    12. Souza DG
    13. Teixeira MM

    (2015) Dengue virus requires the CC-chemokine receptor CCR5 for replication and infection development

    Immunology 145:583–596.

    https://doi.org/10.1111/imm.12476

    • PubMed
    • Google Scholar
48. 1. Melo RCN
    2. Spencer LA
    3. Perez SAC
    4. Neves JS
    5. Bafford SP
    6. Morgan ES
    7. Dvorak AM
    8. Weller PF

    (2009) Vesicle-mediated secretion of human eosinophil granule-derived major basic protein

    Laboratory Investigation; a Journal of Technical Methods and Pathology 89:769–781.

    https://doi.org/10.1038/labinvest.2009.40

    • PubMed
    • Google Scholar
49. 1. Meyts I
    2. Casanova JL

    (2021) Viral infections in humans and mice with genetic deficiencies of the type I IFN response pathway

    European Journal of Immunology 51:1039–1061.

    https://doi.org/10.1002/eji.202048793

    • PubMed
    • Google Scholar
50. 1. Molás RB
    2. Ribeiro MR
    3. Ramalho Dos Santos MJC
    4. Borbely AU
    5. Oliani DV
    6. Oliani AH
    7. Nadkarni S
    8. Nogueira ML
    9. Moreli JB
    10. Oliani SM

    (2020) The involvement of annexin A1 in human placental response to maternal Zika virus infection

    Antiviral Research 179:104809.

    https://doi.org/10.1016/j.antiviral.2020.104809

    • PubMed
    • Google Scholar
51. 1. Morita M
    2. Kuba K
    3. Ichikawa A
    4. Nakayama M
    5. Katahira J
    6. Iwamoto R
    7. Watanebe T
    8. Sakabe S
    9. Daidoji T
    10. Nakamura S
    11. Kadowaki A
    12. Ohto T
    13. Nakanishi H
    14. Taguchi R
    15. Nakaya T
    16. Murakami M
    17. Yoneda Y
    18. Arai H
    19. Kawaoka Y
    20. Penninger JM
    21. Arita M
    22. Imai Y

    (2013) The lipid mediator protectin D1 inhibits influenza virus replication and improves severe influenza

    Cell 153:112–125.

    https://doi.org/10.1016/j.cell.2013.02.027

    • PubMed
    • Google Scholar
52. 1. Morrison J
    2. Rathore APS
    3. Mantri CK
    4. Aman SAB
    5. Nishida A
    6. St John AL

    (2017) Transcriptional Profiling Confirms the Therapeutic Effects of Mast Cell Stabilization in a Dengue Disease Model

    Journal of Virology 91:18.

    https://doi.org/10.1128/JVI.00617-17

    • PubMed
    • Google Scholar
53. 1. Murri M
    2. Insenser M
    3. Bernal-Lopez MR
    4. Perez-Martinez P
    5. Escobar-Morreale HF
    6. Tinahones FJ

    (2013) Proteomic analysis of visceral adipose tissue in pre-obese patients with type 2 diabetes

    Molecular and Cellular Endocrinology 376:99–106.

    https://doi.org/10.1016/j.mce.2013.06.010

    • PubMed
    • Google Scholar
54. 1. Oliveira LG
    2. Souza-Testasicca MC
    3. Vago JP
    4. Figueiredo AB
    5. Canavaci AMC
    6. Perucci LO
    7. Ferreira TPT
    8. Coelho EAF
    9. Gonçalves DU
    10. Rocha MOC
    11. E Silva PMR
    12. Ferreira CN
    13. Queiroz-Junior C
    14. Sousa LP
    15. Fernandes AP

    (2017) Annexin A1 Is Involved in the Resolution of Inflammatory Responses during Leishmania braziliensis Infection

    Journal of Immunology (Baltimore, Md 198:3227–3236.

    https://doi.org/10.4049/jimmunol.1602028

    • PubMed
    • Google Scholar
55. 1. Parisi JDS
    2. Corrêa MP
    3. Gil CD

    (2019) Lack of Endogenous Annexin A1 Increases Mast Cell Activation and Exacerbates Experimental Atopic Dermatitis

    Cells 8:E51.

    https://doi.org/10.3390/cells8010051

    • PubMed
    • Google Scholar
56. 1. Patel HB
    2. Kornerup KN
    3. Sampaio ALF
    4. D’Acquisto F
    5. Seed MP
    6. Girol AP
    7. Gray M
    8. Pitzalis C
    9. Oliani SM
    10. Perretti M

    (2012) The impact of endogenous annexin A1 on glucocorticoid control of inflammatory arthritis

    Annals of the Rheumatic Diseases 71:1872–1880.

    https://doi.org/10.1136/annrheumdis-2011-201180

    • PubMed
    • Google Scholar
57. 1. Perretti M
    2. Ahluwalia A
    3. Harris JG
    4. Goulding NJ
    5. Flower RJ

    (1993)

    Lipocortin-1 fragments inhibit neutrophil accumulation and neutrophil-dependent edema in the mouse. A qualitative comparison with an anti-CD11b monoclonal antibody

    Journal of Immunology (Baltimore, Md 151:4306–4314.

    • PubMed
    • Google Scholar
58. 1. Perretti M
    2. D’Acquisto F

    (2009) Annexin A1 and glucocorticoids as effectors of the resolution of inflammation

    Nature Reviews. Immunology 9:62–70.

    https://doi.org/10.1038/nri2470

    • PubMed
    • Google Scholar
59. 1. Perrone LA
    2. Plowden JK
    3. García-Sastre A
    4. Katz JM
    5. Tumpey TM

    (2008) H5N1 and 1918 pandemic influenza virus infection results in early and excessive infiltration of macrophages and neutrophils in the lungs of mice

    PLOS Pathogens 4:e1000115.

    https://doi.org/10.1371/journal.ppat.1000115

    • PubMed
    • Google Scholar
60. 1. Perucci LO
    2. Sugimoto MA
    3. Gomes KB
    4. Dusse LM
    5. Teixeira MM
    6. Sousa LP

    (2017) Annexin A1 and specialized proresolving lipid mediators: promoting resolution as a therapeutic strategy in human inflammatory diseases

    Expert Opinion on Therapeutic Targets 21:879–896.

    https://doi.org/10.1080/14728222.2017.1364363

    • PubMed
    • Google Scholar
61. 1. Rådinger M
    2. Jensen BM
    3. Swindle E
    4. Gilfillan AM

    (2015) Assay of mast cell mediators

    Methods in Molecular Biology (Clifton, N.J.) 1220:307–323.

    https://doi.org/10.1007/978-1-4939-1568-2_19

    • PubMed
    • Google Scholar
62. 1. Ramon S
    2. Baker SF
    3. Sahler JM
    4. Kim N
    5. Feldsott EA
    6. Serhan CN
    7. Martínez-Sobrido L
    8. Topham DJ
    9. Phipps RP

    (2014) The specialized proresolving mediator 17-HDHA enhances the antibody-mediated immune response against influenza virus: a new class of adjuvant

    Journal of Immunology (Baltimore, Md 193:6031–6040.

    https://doi.org/10.4049/jimmunol.1302795

    • PubMed
    • Google Scholar
63. 1. Rathore AP
    2. Mantri CK
    3. Aman SA
    4. Syenina A
    5. Ooi J
    6. Jagaraj CJ
    7. Goh CC
    8. Tissera H
    9. Wilder-Smith A
    10. Ng LG
    11. Gubler DJ
    12. St John AL

    (2019) Dengue virus-elicited tryptase induces endothelial permeability and shock

    The Journal of Clinical Investigation 129:4180–4193.

    https://doi.org/10.1172/JCI128426

    • PubMed
    • Google Scholar
64. 1. Rathore APS
    2. Senanayake M
    3. Athapathu AS
    4. Gunasena S
    5. Karunaratna I
    6. Leong WY
    7. Lim T
    8. Mantri CK
    9. Wilder-Smith A
    10. St John AL

    (2020) Serum chymase levels correlate with severe dengue warning signs and clinical fluid accumulation in hospitalized pediatric patients

    Scientific Reports 10:11856.

    https://doi.org/10.1038/s41598-020-68844-z

    • PubMed
    • Google Scholar
65. 1. Rowe EK
    2. Leo Y-S
    3. Wong JGX
    4. Thein T-L
    5. Gan VC
    6. Lee LK
    7. Lye DC

    (2014) Challenges in dengue fever in the elderly: atypical presentation and risk of severe dengue and hospital-acquired infection

    PLOS Neglected Tropical Diseases 8:e2777.

    https://doi.org/10.1371/journal.pntd.0002777

    • PubMed
    • Google Scholar
66. 1. Sangkaew S
    2. Ming D
    3. Boonyasiri A
    4. Honeyford K
    5. Kalayanarooj S
    6. Yacoub S
    7. Dorigatti I
    8. Holmes A

    (2021) Risk predictors of progression to severe disease during the febrile phase of dengue: a systematic review and meta-analysis

    The Lancet. Infectious Diseases 21:1014–1026.

    https://doi.org/10.1016/S1473-3099(20)30601-0

    • PubMed
    • Google Scholar
67. 1. Saria A
    2. Lundberg JM

    (1983) Evans blue fluorescence: quantitative and morphological evaluation of vascular permeability in animal tissues

    Journal of Neuroscience Methods 8:41–49.

    https://doi.org/10.1016/0165-0270(83)90050-x

    • PubMed
    • Google Scholar
68. 1. Schett G
    2. Neurath MF

    (2018) Resolution of chronic inflammatory disease: universal and tissue-specific concepts

    Nature Communications 9:3261.

    https://doi.org/10.1038/s41467-018-05800-6

    • PubMed
    • Google Scholar
69. 1. Schloer S
    2. Hübel N
    3. Masemann D
    4. Pajonczyk D
    5. Brunotte L
    6. Ehrhardt C
    7. Brandenburg L-O
    8. Ludwig S
    9. Gerke V
    10. Rescher U

    (2019) The annexin A1/FPR2 signaling axis expands alveolar macrophages, limits viral replication, and attenuates pathogenesis in the murine influenza A virus infection model

    FASEB Journal 33:12188–12199.

    https://doi.org/10.1096/fj.201901265R

    • PubMed
    • Google Scholar
70. 1. Sekheri M
    2. El Kebir D
    3. Edner N
    4. Filep JG

    (2020) 15-Epi-LXA4 and 17-epi-RvD1 restore TLR9-mediated impaired neutrophil phagocytosis and accelerate resolution of lung inflammation

    PNAS 117:7971–7980.

    https://doi.org/10.1073/pnas.1920193117

    • PubMed
    • Google Scholar
71. 1. Sena A
    2. Grishina I
    3. Thai A
    4. Goulart L
    5. Macal M
    6. Fenton A
    7. Li J
    8. Prindiville T
    9. Oliani SM
    10. Dandekar S
    11. Goulart L
    12. Sankaran-Walters S

    (2013) Dysregulation of anti-inflammatory annexin A1 expression in progressive Crohns Disease

    PLOS ONE 8:e76969.

    https://doi.org/10.1371/journal.pone.0076969

    • PubMed
    • Google Scholar
72. 1. Senchenkova EY
    2. Ansari J
    3. Becker F
    4. Vital SA
    5. Al-Yafeai Z
    6. Sparkenbaugh EM
    7. Pawlinski R
    8. Stokes KY
    9. Carroll JL
    10. Dragoi A-M
    11. Qin CX
    12. Ritchie RH
    13. Sun H
    14. Cuellar-Saenz HH
    15. Rubinstein MR
    16. Han YW
    17. Orr AW
    18. Perretti M
    19. Granger DN
    20. Gavins FNE

    (2019) Novel Role for the AnxA1-Fpr2/ALX Signaling Axis as a Key Regulator of Platelet Function to Promote Resolution of Inflammation

    Circulation 140:319–335.

    https://doi.org/10.1161/CIRCULATIONAHA.118.039345

    • PubMed
    • Google Scholar
73. 1. Sheikh MH
    2. Solito E

    (2018) Annexin A1: Uncovering the Many Talents of an Old Protein

    International Journal of Molecular Sciences 19:E1045.

    https://doi.org/10.3390/ijms19041045

    • PubMed
    • Google Scholar
74. 1. Shepard DS
    2. Undurraga EA
    3. Halasa YA
    4. Stanaway JD

    (2016) The global economic burden of dengue: a systematic analysis

    The Lancet. Infectious Diseases 16:935–941.

    https://doi.org/10.1016/S1473-3099(16)00146-8

    • PubMed
    • Google Scholar
75. 1. Shirey KA
    2. Lai W
    3. Pletneva LM
    4. Karp CL
    5. Divanovic S
    6. Blanco JCG
    7. Vogel SN

    (2014) Role of the lipoxygenase pathway in RSV-induced alternatively activated macrophages leading to resolution of lung pathology

    Mucosal Immunology 7:549–557.

    https://doi.org/10.1038/mi.2013.71

    • PubMed
    • Google Scholar
76. 1. Shresta S
    2. Kyle JL
    3. Snider HM
    4. Basavapatna M
    5. Beatty PR
    6. Harris E

    (2004) Interferon-dependent immunity is essential for resistance to primary dengue virus infection in mice, whereas T- and B-cell-dependent immunity are less critical

    Journal of Virology 78:2701–2710.

    https://doi.org/10.1128/jvi.78.6.2701-2710.2004

    • PubMed
    • Google Scholar
77. 1. Sinniah A
    2. Yazid S
    3. Perretti M
    4. Solito E
    5. Flower RJ

    (2016) The role of the Annexin-A1/FPR2 system in the regulation of mast cell degranulation provoked by compound 48/80 and in the inhibitory action of nedocromil

    International Immunopharmacology 32:87–95.

    https://doi.org/10.1016/j.intimp.2016.01.003

    • PubMed
    • Google Scholar
78. 1. Sinniah A
    2. Yazid S
    3. Bena S
    4. Oliani SM
    5. Perretti M
    6. Flower RJ

    (2019) Endogenous Annexin-A1 Negatively Regulates Mast Cell-Mediated Allergic Reactions

    Frontiers in Pharmacology 10:1313.

    https://doi.org/10.3389/fphar.2019.01313

    • PubMed
    • Google Scholar
79. 1. Solito E
    2. Mulla A
    3. Morris JF
    4. Christian HC
    5. Flower RJ
    6. Buckingham JC

    (2003) Dexamethasone induces rapid serine-phosphorylation and membrane translocation of annexin 1 in a human folliculostellate cell line via a novel nongenomic mechanism involving the glucocorticoid receptor, protein kinase C, phosphatidylinositol 3-kinase, and mitogen-activated protein kinase

    Endocrinology 144:1164–1174.

    https://doi.org/10.1210/en.2002-220592

    • PubMed
    • Google Scholar
80. 1. Souza DG
    2. Fagundes CT
    3. Sousa LP
    4. Amaral FA
    5. Souza RS
    6. Souza AL
    7. Kroon EG
    8. Sachs D
    9. Cunha FQ
    10. Bukin E
    11. Atrasheuskaya A
    12. Ignatyev G
    13. Teixeira MM

    (2009) Essential role of platelet-activating factor receptor in the pathogenesis of Dengue virus infection

    PNAS 106:14138–14143.

    https://doi.org/10.1073/pnas.0906467106

    • PubMed
    • Google Scholar
81. 1. Spite M
    2. Norling LV
    3. Summers L
    4. Yang R
    5. Cooper D
    6. Petasis NA
    7. Flower RJ
    8. Perretti M
    9. Serhan CN

    (2009) Resolvin D2 is a potent regulator of leukocytes and controls microbial sepsis

    Nature 461:1287–1291.

    https://doi.org/10.1038/nature08541

    • PubMed
    • Google Scholar
82. 1. St John AL
    2. Rathore APS
    3. Yap H
    4. Ng ML
    5. Metcalfe DD
    6. Vasudevan SG
    7. Abraham SN

    (2011) Immune surveillance by mast cells during dengue infection promotes natural killer (NK) and NKT-cell recruitment and viral clearance

    PNAS 108:9190–9195.

    https://doi.org/10.1073/pnas.1105079108

    • PubMed
    • Google Scholar
83. 1. St John AL
    2. Abraham SN
    3. Gubler DJ

    (2013a) Barriers to preclinical investigations of anti-dengue immunity and dengue pathogenesis

    Nature Reviews. Microbiology 11:420–426.

    https://doi.org/10.1038/nrmicro3030

    • PubMed
    • Google Scholar
84. 1. St John AL
    2. Rathore APS
    3. Raghavan B
    4. Ng ML
    5. Abraham SN

    (2013b) Contributions of mast cells and vasoactive products, leukotrienes and chymase, to dengue virus-induced vascular leakage

    eLife 2:e00481.

    https://doi.org/10.7554/eLife.00481

    • PubMed
    • Google Scholar
85. 1. Stanaway JD
    2. Shepard DS
    3. Undurraga EA
    4. Halasa YA
    5. Coffeng LE
    6. Brady OJ
    7. Hay SI
    8. Bedi N
    9. Bensenor IM
    10. Castañeda-Orjuela CA
    11. Chuang T-W
    12. Gibney KB
    13. Memish ZA
    14. Rafay A
    15. Ukwaja KN
    16. Yonemoto N
    17. Murray CJL

    (2016) The global burden of dengue: an analysis from the Global Burden of Disease Study 2013

    The Lancet. Infectious Diseases 16:712–723.

    https://doi.org/10.1016/S1473-3099(16)00026-8

    • PubMed
    • Google Scholar
86. 1. Sugimoto MA
    2. Vago JP
    3. Teixeira MM
    4. Sousa LP

    (2016) Annexin A1 and the Resolution of Inflammation: Modulation of Neutrophil Recruitment, Apoptosis, and Clearance

    Journal of Immunology Research 2016:8239258.

    https://doi.org/10.1155/2016/8239258

    • PubMed
    • Google Scholar
87. 1. Sugimoto MA
    2. Vago JP
    3. Perretti M
    4. Teixeira MM

    (2019) Mediators of the Resolution of the Inflammatory Response

    Trends in Immunology 40:212–227.

    https://doi.org/10.1016/j.it.2019.01.007

    • PubMed
    • Google Scholar
88. 1. Syenina A
    2. Jagaraj CJ
    3. Aman SAB
    4. Sridharan A
    5. St John AL

    (2015) Dengue vascular leakage is augmented by mast cell degranulation mediated by immunoglobulin Fcgamma receptors

    eLife 4:e05291.

    https://doi.org/10.7554/eLife.05291

    • PubMed
    • Google Scholar
89. 1. Tavares LP
    2. Garcia CC
    3. Vago JP
    4. Queiroz-Junior CM
    5. Galvão I
    6. David BA
    7. Rachid MA
    8. Silva PMR
    9. Russo RC
    10. Teixeira MM
    11. Sousa LP

    (2016) Inhibition of Phosphodiesterase-4 during Pneumococcal Pneumonia Reduces Inflammation and Lung Injury in Mice

    American Journal of Respiratory Cell and Molecular Biology 55:24–34.

    https://doi.org/10.1165/rcmb.2015-0083OC

    • PubMed
    • Google Scholar
90. 1. The Lancet Infectious Diseases

    (2018) The dengue vaccine dilemma

    The Lancet. Infectious Diseases 18:123.

    https://doi.org/10.1016/S1473-3099(18)30023-9

    • PubMed
    • Google Scholar
91. 1. Thul S
    2. Labat C
    3. Temmar M
    4. Benetos A
    5. Bäck M

    (2017) Low salivary resolvin D1 to leukotriene B4 ratio predicts carotid intima media thickness: A novel biomarker of non-resolving vascular inflammation

    European Journal of Preventive Cardiology 24:903–906.

    https://doi.org/10.1177/2047487317694464

    • PubMed
    • Google Scholar
92. 1. Tissera H
    2. Rathore APS
    3. Leong WY
    4. Pike BL
    5. Warkentien TE
    6. Farouk FS
    7. Syenina A
    8. Eong Ooi E
    9. Gubler DJ
    10. Wilder-Smith A
    11. St John AL

    (2017) Chymase Level Is a Predictive Biomarker of Dengue Hemorrhagic Fever in Pediatric and Adult Patients

    The Journal of Infectious Diseases 216:1112–1121.

    https://doi.org/10.1093/infdis/jix447

    • PubMed
    • Google Scholar
93. 1. Tzelepis F
    2. Verway M
    3. Daoud J
    4. Gillard J
    5. Hassani-Ardakani K
    6. Dunn J
    7. Downey J
    8. Gentile ME
    9. Jaworska J
    10. Sanchez AMJ
    11. Nédélec Y
    12. Vali H
    13. Tabrizian M
    14. Kristof AS
    15. King IL
    16. Barreiro LB
    17. Divangahi M

    (2015) Annexin1 regulates DC efferocytosis and cross-presentation during Mycobacterium tuberculosis infection

    The Journal of Clinical Investigation 125:752–768.

    https://doi.org/10.1172/JCI77014

    • PubMed
    • Google Scholar
94. 1. Vago JP
    2. Nogueira CRC
    3. Tavares LP
    4. Soriani FM
    5. Lopes F
    6. Russo RC
    7. Pinho V
    8. Teixeira MM
    9. Sousa LP

    (2012) Annexin A1 modulates natural and glucocorticoid-induced resolution of inflammation by enhancing neutrophil apoptosis

    Journal of Leukocyte Biology 92:249–258.

    https://doi.org/10.1189/jlb.0112008

    • PubMed
    • Google Scholar
95. 1. Vanessa KHQ
    2. Julia MG
    3. Wenwei L
    4. Michelle ALT
    5. Zarina ZRS
    6. Lina LHK
    7. Sylvie A

    (2015) Absence of Annexin A1 impairs host adaptive immunity against Mycobacterium tuberculosis in vivo

    Immunobiology 220:614–623.

    https://doi.org/10.1016/j.imbio.2014.12.001

    • PubMed
    • Google Scholar
96. 1. Vong L
    2. Ferraz JGP
    3. Dufton N
    4. Panaccione R
    5. Beck PL
    6. Sherman PM
    7. Perretti M
    8. Wallace JL

    (2012) Up-regulation of Annexin-A1 and lipoxin A(4) in individuals with ulcerative colitis may promote mucosal homeostasis

    PLOS ONE 7:e39244.

    https://doi.org/10.1371/journal.pone.0039244

    • PubMed
    • Google Scholar
97. Book

    1. WHO

    (2009)

    Dengue Guidelines for Diagnosis, Treatment, Prevention and Control (new edition)

    World Health Organization.

    • Google Scholar
98. 1. WHO

    (2018)

    Dengue vaccine: WHO position paper – September 2018

    Weekly Epidemiological Record 36:457–476.

    • Google Scholar
99. 1. Williams KL
    2. Zompi S
    3. Beatty PR
    4. Harris E

    (2009) A mouse model for studying dengue virus pathogenesis and immune response

    Annals of the New York Academy of Sciences 1171 Suppl 1:E12–E23.

    https://doi.org/10.1111/j.1749-6632.2009.05057.x

    • PubMed
    • Google Scholar
100. 1. Xu X
     2. Gao W
     3. Li L
     4. Hao J
     5. Yang B
     6. Wang T
     7. Li L
     8. Bai X
     9. Li F
     10. Ren H
     11. Zhang M
     12. Zhang L
     13. Wang J
     14. Wang D
     15. Zhang J
     16. Jiao L

     (2021) Annexin A1 protects against cerebral ischemia-reperfusion injury by modulating microglia/macrophage polarization via FPR2/ALX-dependent AMPK-mTOR pathway

     Journal of Neuroinflammation 18:119.

     https://doi.org/10.1186/s12974-021-02174-3

     • PubMed
     • Google Scholar
101. 1. Yazid S
     2. Sinniah A
     3. Solito E
     4. Calder V
     5. Flower RJ

     (2013) Anti-allergic cromones inhibit histamine and eicosanoid release from activated human and murine mast cells by releasing Annexin A1

     PLOS ONE 8:e58963.

     https://doi.org/10.1371/journal.pone.0058963

     • PubMed
     • Google Scholar
102. 1. Zandi K
     2. Bassit L
     3. Amblard F
     4. Cox BD
     5. Hassandarvish P
     6. Moghaddam E
     7. Yueh A
     8. Libanio Rodrigues GO
     9. Passos I
     10. Costa VV
     11. AbuBakar S
     12. Zhou L
     13. Kohler J
     14. Teixeira MM
     15. Schinazi RF

     (2019) Nucleoside Analogs with Selective Antiviral Activity against Dengue Fever and Japanese Encephalitis Viruses

     Antimicrobial Agents and Chemotherapy 63:e00397-19.

     https://doi.org/10.1128/AAC.00397-19

     • PubMed
     • Google Scholar

Decision letter after peer review:

Thank you for submitting your work entitled "Targeting the Annexin A1-FPR2/ALX pathway for host-directed therapy in dengue disease" for further consideration by eLife. Your article has been reviewed by 3 peer reviewers, including Frank Hirchhoff as Reviewing Editor and Reviewer #1, and the evaluation has been evaluated by Jos Van der Meer as Senior Editor. The reviewers have opted to remain anonymous.

There are some significant remaining issues that need to be addressed, as outlined below.

Essential revisions:

1. The improvements in disease symptoms appeared to be independent of the viral load. Since AC2-26 had previously been shown to have anti-inflammatory properties these findings are, while intriguing in the context of disease, not necessarily novel. The authors should mention that corticosteroids have been previously examined for the treatment of dengue disease and may exert beneficial effects through the induction of Annexin A1 and ALXR expression. They should also discuss why FPR2/ALX agonists may be better than treatment with corticosteroids to suppress exaggerated inflammation.

2. It is well established that DENV does not replicate efficiently in mice, but infection can occur using mouse-adapted strains of DENV and/or immunocompromised mouse strains. Here, the authors use a combination of wt (Balb/C and C57BL/6) and mouse mutant lines (including AG129 mice which have been shown to support robust infection with mouse adapted DENV strains) and infect them with a very high dose of DENV2 isolate. There are very limited viremia data shown throughout the manuscript which makes is somewhat difficult to interpret some of the results. How much are the effects in all the non-AG129 mouse lines attributable to an acute response to the high inoculum (and no/very limited replication) vs actual infection?

3. In Figure 4, when using IFNα/βR-/- mice (AG129), IL-6 and CCL5 instead of MCPT-1 and CCL2 were used as markers of systemic inflammatory mediators, which is not consistent with the previous experiments done in immunocompetent mice. No results of MCPT-1 and CCL2 changes in AG129 mice was presented and the authors didn't explain why they changed the markers in this manuscript. Since MCPT-1 is a mast cell-specific product, if it is not decreased in AG129 mice treated with Ac2-26 after DENV infection, how would the authors explain the protective effects of Ac2-26 in AG129 mice in the context of DENV infection?

4. The authors showed AnxA1 memetic peptide Ac2-26 could suppress MC degranulation. However, the same experiments should also be done in parallel in AnxA1-/- and FPR2/ALX-/- mice and in BMMCs derived from them. If Ac2-26 can prevent MC degranulation, it is expected to see decreased degranulation in AnxA1 mice but not FPR2/ALX KO mice when infected with DENV, both in vivo and in BMMCs. Finally, the peptide concentrations in mice (150 µg/animal) and in vitro (100 µM) are pretty high. The authors should mention whether any side effects were observed.

Reviewer #1 (Recommendations for the authors):

Dengue virus infection is spread by mosquitos and typically causes high fever, headache, vomiting, muscle and joint pains, and a characteristic skin rash. Typically, these symptoms resolved within a week but in some cases, dengue disease becomes more severe and is leads to hemorrhagic fever or dengue shock syndrome. Currently, no specific treatment against dengue disease is available. In the present study, Costa and colleagues hypothesized that excessive inflammatory responses play a key role in severe dengue disease and that anti-inflammatory molecules may be reduced in infected individuals. Specifically, they show that the circulating levels of anti-inflammatory Annexin A1 are reduced in both dengue patients and in infected mice. They further report that lack of Annexin A1 or its receptor FPR2/ALX enhance the disease in infected mice, while treatment with Annexin A1 agonistic peptide had beneficial effects. They further propose that the clinical outcome is dependent on inflammation and mast cell activation rather than the levels of viral replication. The authors conclude that Annexin A1 plays a key role in the clinical outcome of dengue virus infection and that FPR2/ALX agonists may offer therapeutic perspectives. Strength of the present study are that the authors analyzed both patient as well as mouse sample. Studies in knockout animals, provide strong evidence that the attenuating effect of Annexin A1 on the inflammatory response indeed attenuates the severity of dengue disease. Additional strengths are the evidence that an agonistic peptide exerts protective effect. A limitation is that the concentrations of the peptide used to monitor its effects (100 µM) are pretty high. In addition, the treatment is not specific and the concept of utilizing anti-inflammatory agents is not novel. Previous studies suggested that corticosteroids may prevent the progression to severe dengue disease (Rathnasiri Bandaraa and Herathb, Heliyon, 2018). Corticosteroids exert their anti-inflammatory effects by the induction of Annexin A1 and ALXR expression. This link should be mentioned and its needs to be explained why FPR2/ALX agonists may be better than treatment with corticosteroids to suppress exaggerated inflammation.

1. The authors should mention that corticosteroids have been previously examined for the treatment of dengue disease and may exert beneficial effects through the induction of Annexin A1 and ALXR expression. They should also discuss why FPR2/ALX agonists may be better than treatment with corticosteroids to suppress exaggerated inflammation.

2. Some references seem flawed (e.g. line 53, The Lancet Infectious, 2018). Should be checked throughout.

3. Line 254: "Groups were comparable for sex distribution". Table 1 shows that the proportion of females was higher in the control group and that, on average, age was higher in patients with severe dengue disease. This should be mentioned in the text.

4. It would be interesting to know whether the levels of Annexin A1 in individuals with dengue infection (Figure 1A) are inversely correlated with the levels of inflammatory cytokines.

5. The peptide concentrations in mice (150 µg/animal) and in vitro (100 µM) are pretty high. The authors should mention whether any side effects were observed.

6. The Annexin A1 levels in patients were about 10-fold lower than in mice (Figure 1). This difference should be discussed.

7. The authors mention that treatment with the Ac2-26 peptide did not affect systemic viral burden. It seems somewhat surprising that attenuation of innate immune activation doesn't affect the levels of viral replication. Please discuss.

Reviewer #2 (Recommendations for the authors):

In this manuscript, Costa et al., investigated the role of AnxA1-FPR2/ALX pathway in disease severity of dengue virus (DENV) infection and explores the therapeutic benefits of AnxA1 memetic peptide in DENV infection using mouse model. The authors show that serum levels of pro-resolving protein AnxA1 are significantly decreased in patients with DENV infection and infected mice. They show that the absence of AnxA-FPR2/ALX pathway in mice lead to aggravated dengue disease parameters by measuring hematological parameters, vascular leakage, and inflammatory factors. They further demonstrate that administration of a AxnA1 memetic peptide, Ac2-26, could improve these disease parameters in WT, AnxA1-/-, but not mice lacking the AnxA1 receptor, FPR2/ALX. In IFNα/βR-/- mice which are more susceptible to DENV infection, treatment of Ac2-26 improved the clinical outcome along with the disease parameters. The authors finally expore the mechanism of Ac2-26's protective effects and showing that Ac2-26 prevents mast cell degranulation both in vitro and in vivo models.

Overall, this study provides new insights on the mechanisms that underlie severe dengue disease. Notably, the data presented here suggest that the AnxA1-FPR2/ALX pathway may be a suitable potential target for host-directed therapy and a biomarker for stratifying the severity of in human dengue disease. The design of the experiments is largely adequate and has yielded data are mostly supportive of the conclusions drawn here.

Major issues that affect impact:

– The improvements in disease symptoms appeared to be independent of the viral load. Since AC2-26 had previously been shown to have anti-inflammatory properties these findings are, while intriguing in the context of disease, not necessarily novel.

– It is well established that DENV does not replicate efficiently in mice, but infection can occur using mouse-adapted strains of DENV and/or immunocompromised mouse strains. Here, the authors use a combination of wt (Balb/C and C57BL/6) and mouse mutant lines (including AG129 mice which have been shown to support robust infection with mouse adapted DENV strains) and infect them with a very high dose of DENV2 isolate. There are very limited viremia data shown throughout the manuscript which makes is somewhat difficult to interpret some of the results. How much are the effects in all the non-AG129 mouse lines attributable to an acute response to the high inoculum (and no/very limited replication) vs actual infection?

– In Figure 4, when using IFNα/βR-/- mice (AG129), IL-6 and CCL5 instead of MCPT-1 and CCL2 were used as markers of systemic inflammatory mediators, which is not consistent with the previous experiments done in immunocompetent mice. No results of MCPT-1 and CCL2 changes in AG129 mice was presented and the authors didn't explain why they changed the markers in this manuscript. Since MCPT-1 is a mast cell-specific product, if it is not decreased in AG129 mice treated with Ac2-26 after DENV infection, how would the authors explain the protective effects of Ac2-26 in AG129 mice in the context of DENV infection?

– The authors showed AnxA1 memetic peptide Ac2-26 could suppress MC degranulation. However, the same experiments should also be done in parallel in AnxA1-/- and FPR2/ALX-/- mice and in BMMCs derived from them. If Ac2-26 can prevent MC degranulation, it is expected to see decreased degranulation in AnxA1 mice but not FPR2/ALX KO mice when infected with DENV, both in vivo and in BMMCs.

Reviewer #3 (Recommendations for the authors):

Costa and colleagues investigated the involvement of the pro-resolving protein Annexin A1 (AnxA1) in controlling vascular leakage and cytokine storm associated with dengue disease most severe forms. They find a decrease of AnxA1 levels in the plasma of patients during acute dengue virus infection compared to healthy patients. This decrease is even more pronounced in patients with severe dengue, i.e. requiring hospitalization, compared to those with non-severe dengue. A similar AnxA1 profile was measured over time in dengue-infected wild-type mice of two different strains and further investigated in this model. Animals lacking AnxA1 or AnxA1-receptor FPR2/ALX showed more severe pathology than wild-type mice, while maintaining unchanged viral load. Importantly, the administration of an AnxA1 mimetic peptide strongly attenuated the manifestations of dengue disease in wild-type and AnxA1 knockout mice. This effect could be explained in part by the limitation of mast cell degranulation in vivo and in vitro.

The strength of the article lies in the combination of observations made in patients with reproducible phenotypes in different mouse models, as well as in the testing of molecules with therapeutic potential. The manuscript is scientifically sound and the experiments are well executed. In general, the data are of very good quality, well analyzed and clearly presented. This study provides a very interesting perspective on the pharmacology of resolution, highlighting the benefit of expanding the scope of therapies in the future beyond that of viral load control in order to limit the overshoot of immune responses and associated damage.

Essential revisions:

1. The improvements in disease symptoms appeared to be independent of the viral load. Since AC2-26 had previously been shown to have anti-inflammatory properties these findings are, while intriguing in the context of disease, not necessarily novel.

We have addressed this valid point in our reply to Reviewer #2 (comment #1). The relevance and novelty of our findings were further highlighted in the updated version of our manuscript.

The authors should mention that corticosteroids have been previously examined for the treatment of dengue disease and may exert beneficial effects through the induction of Annexin A1 and ALXR expression. They should also discuss why FPR2/ALX agonists may be better than treatment with corticosteroids to suppress exaggerated inflammation.

We have addressed this important point in our reply to Reviewer #1 (comment #1). Following the Reviewer's advice, the revised manuscript includes a discussion highlighting (i) the potential involvement of AnxA1 in the beneficial actions of corticosteroids in dengue disease and (ii) the therapeutic benefit of targeting this pathway over established anti-inflammatory therapies.

2. It is well established that DENV does not replicate efficiently in mice, but infection can occur using mouse-adapted strains of DENV and/or immunocompromised mouse strains. Here, the authors use a combination of wt (Balb/C and C57BL/6) and mouse mutant lines (including AG129 mice which have been shown to support robust infection with mouse adapted DENV strains) and infect them with a very high dose of DENV2 isolate. There are very limited viremia data shown throughout the manuscript which makes is somewhat difficult to interpret some of the results. How much are the effects in all the non-AG129 mouse lines attributable to an acute response to the high inoculum (and no/very limited replication) vs actual infection?

We have carefully addressed this concern in our reply to Reviewer #2 (comment #2) and included a new paragraph in the revised manuscript discussing this important point.

3. In Figure 4, when using IFNα/βR-/- mice (AG129), IL-6 and CCL5 instead of MCPT-1 and CCL2 were used as markers of systemic inflammatory mediators, which is not consistent with the previous experiments done in immunocompetent mice. No results of MCPT-1 and CCL2 changes in AG129 mice was presented and the authors didn't explain why they changed the markers in this manuscript. Since MCPT-1 is a mast cell-specific product, if it is not decreased in AG129 mice treated with Ac2-26 after DENV infection, how would the authors explain the protective effects of Ac2-26 in AG129 mice in the context of DENV infection?

As suggested by the Reviewer, we have included data on the systemic levels of MCPT-1 and CCL2 in A129 mice subjected to the model of severe dengue (Figure 4H,I). Please, refer to our reply to Reviewer #2 (comment #3).

4. The authors showed AnxA1 memetic peptide Ac2-26 could suppress MC degranulation. However, the same experiments should also be done in parallel in AnxA1-/- and FPR2/ALX-/- mice and in BMMCs derived from them. If Ac2-26 can prevent MC degranulation, it is expected to see decreased degranulation in AnxA1 mice but not FPR2/ALX KO mice when infected with DENV, both in vivo and in BMMCs.

As suggested by the Reviewer, we include new data in the revised version of the manuscript showing the dependence of FPR2/ALX for the modulatory effect of Ac_2-26 in mast cell degranulation (Figure 6B,C).

Finally, the peptide concentrations in mice (150 µg/animal) and in vitro (100 µM) are pretty high. The authors should mention whether any side effects were observed.

As detailed in our reply to Reviewer #1 (comment #5), the in vivo dose used in our study was based on previous publications from our group and others (Damazo, Yona, Flower, Perretti, and Oliani, 2006; Galvao et al., 2017; Machado et al., 2020; Perretti, Ahluwalia, Harris, Goulding, and Flower, 1993; Vago et al., 2012). Using 150 µg/animal, we did not observe any notable adverse effects in our experiments (we have included this observation in the revised text).

Our in vitro concentration of Ac_2-26 is consistent with previous studies on mast cells, where the prevention of mast cell degranulation is only achieved with high peptide concentrations (M. P. Oliveira, Prates, Gimenes, Correa, and Oliani, 2021). Besides, the concentration of Ac_2-26 used in our in vitro studies was based on a concentration-response performed in bone-marrow-derived mast cells (Figure 6D of the updated manuscript).

Reviewer #1 (Recommendations for the authors):

1. The authors should mention that corticosteroids have been previously examined for the treatment of dengue disease and may exert beneficial effects through the induction of Annexin A1 and ALXR expression. They should also discuss why FPR2/ALX agonists may be better than treatment with corticosteroids to suppress exaggerated inflammation.

We thank the Reviewer for their insightful comment. Although guidelines from the World Health Organization for managing dengue fever only provide recommendations on fluid management and supportive care (WHO, ‎2009), the use of anti-inflammatories in dengue disease has been evaluated. While the use of non-steroidal anti-inflammatory agents in dengue is discouraged, as they can increase the risk of bleeding (PAHO, 2016; WHO, ‎2009), recent studies have pointed towards a benefit of using corticoids in severe dengue with no significant adverse consequences or promotion of virus replication (S. M. R. Bandara and Herath, 2018; S. M. Rathnasiri Bandara and Herath, 2020). Noteworthy, amongst the several genomic and non-genomic actions of glucocorticoids, promotion of AnxA1 has shown to be relevant to the benefits afforded by steroids (De Caterina et al., 1993; Solito et al., 2003), as suggested by the loss of the therapeutic effect of glucocorticoids on AnxA1 depleted animals (Damazo et al., 2006; Patel et al., 2012; Perretti and D'Acquisto, 2009; Vago et al., 2012).

Our study reveals a central role of the axis AnxA1-FPR2/ALX in the pathogenesis of dengue disease. We suggest that this critical anti-inflammatory/pro-resolving pathway is downregulated during dengue, contributing to unbalanced inflammation and the cytokine storm characteristic of the disease. Therefore, there is scope to hypothesise that the benefit of corticosteroids in dengue disease could be partially attributed to promoting the glucocorticoid-inducible Annexin A1/FPR2/ALX pathway. Further investigations should confirm AnxA1 as an underlying mechanism of corticosteroids in dengue disease.

The effectiveness of the treatment with AnxA1 peptide in our study suggests that glucocorticoid therapy could be refined using a more targeted approach, particularly in infectious diseases where there is an underlying imbalance in the AnxA1 pathway. This opens an opportunity to explore small molecules targeting FPR2/ALX receptors, avoiding the spanning adverse consequences of glucocorticoids while targeting the defect in inflammation resolution operating in the disease settings. Further investigation on GC and GC-inducible AnxA1 is particularly relevant in dengue disease, where non-steroidal anti-inflammatory agents are discouraged, as mentioned above.

The revised manuscript includes a discussion highlighting (i) the potential involvement of AnxA1 in the beneficial actions of corticosteroids in dengue disease and (ii) the therapeutic benefit of targeting this pathway over established anti-inflammatory therapies (lines 620-640; 645).

2. Some references seem flawed (e.g. line 53, The Lancet Infectious, 2018). Should be checked throughout.

Thank you for noting this. We have replaced the reference in line 53 and thoroughly revised the other references in the manuscript.

3. Line 254: "Groups were comparable for sex distribution". Table 1 shows that the proportion of females was higher in the control group and that, on average, age was higher in patients with severe dengue disease. This should be mentioned in the text.

Thank you for your comment. Higher age in severe dengue cases is expected since older age is a risk factor for progression to severe disease (Rowe et al., 2014; Sangkaew et al., 2021). We have highlighted this age distribution in the updated version of our manuscript (line 263). As depicted in Table 1 legend, the sex ratio was statistically similar between the groups (p>0.1), making the groups comparable for sex distribution.

4. It would be interesting to know whether the levels of Annexin A1 in individuals with dengue infection (Figure 1A) are inversely correlated with the levels of inflammatory cytokines.

Although we agree with the Reviewer that this could be an interesting point to be addressed, we do not hold additional plasma samples to perform further ELISA assays. However, our research group is conducting an in-depth analysis of imbalances between inflammatory and pro-resolving mediators in plasma samples from dengue patients that will be comprehensively covered in a future publication.

Of note, in both models of dengue disease (immunocompetent and IFNα/βR KO animals), we show that decreased levels of AnxA1 during DENV infection are mirrored by higher levels of inflammatory markers, including the mast cell-specific product.

5. The peptide concentrations in mice (150 µg/animal) and in vitro (100 µM) are pretty high. The authors should mention whether any side effects were observed.

The dose of Ac_2-26 peptide (150μg/animal) was based on previous publications, such as those in the settings of peritonitis (Damazo et al., 2006), gout (Galvao et al., 2017), air pouch (Perretti et al., 1993), pleurisy (Vago et al., 2012) and lung infection (Machado et al., 2020). Initial studies on Ac_2-26 have used a dose of 200 µg per animal to prevent leukocyte migration induced by IL-1β, with no reports of adverse effects (Perretti et al., 1993). In addition, we did not observe any notable adverse effect in our experiments, as mentioned in line 390 of the updated manuscript.

Our in vitro concentration of Ac_2-26 is consistent with previous studies on mast cells (cell lines, BMMC and subcutaneous tissue fragments), where prevention of mast cell degranulation is only achieved with a concentration in the order of micrograms per millilitre (M. P. Oliveira et al., 2021). To confirm the requirement of a high concentration of Ac_2-26 in vitro, we performed a concentration-response, as depicted in Figure 6D of the updated manuscript.

6. The Annexin A1 levels in patients were about 10-fold lower than in mice (Figure 1). This difference should be discussed.

We thank the Reviewer for their comment. The plasma levels observed in our study are in line with previous measurements performed with a variety of studies using homemade ELISA assays or commercial ELISA kits (Adel et al., 2020; Purvis et al., 2019; Santana et al., 2021). Significant differences between animal and human samples in the same study were noticed before (Senchenkova et al., 2019; Xu et al., 2021). We have added this discussion in line 284 of the manuscript.

7. The authors mention that treatment with the Ac2-26 peptide did not affect systemic viral burden. It seems somewhat surprising that attenuation of innate immune activation doesn't affect the levels of viral replication. Please discuss.

This is an important point that has been discussed throughout the manuscript and further highlighted in the revised document (lines 89; 623; 648).

The ability of pro-resolving drugs to balance inflammation without hampering the host's ability to deal with pathogens is, in fact, a hallmark of resolution pharmacology, representing an advantage compared to standard anti-inflammatory drugs (Perretti, Leroy, Bland, and Montero-Melendez, 2015).

Several studies have suggested that pro-resolving mediators can suppress inflammation and diminish inflammatory responses while improving the phagocytosis of pathogens. This was initially demonstrated for specialised lipid mediators in bacterial infection (Chiang et al., 2012; Decker, Sadhu, and Fredman, 2021; Sekheri, El Kebir, Edner, and Filep, 2020), with subsequent studies with Annexin A1 on various infection models (Gobbetti et al., 2014; Machado et al., 2020; L. G. Oliveira et al., 2017; Tzelepis et al., 2015; Vanessa et al., 2015). Therefore, it is already well accepted that pro-resolving drugs can promote innate immunity against bacterial pathogens while reducing the deleterious effects of inflammation. Here we translate these findings into a model of infection. Please refer to our reply to Reviewer #2 (comment 1), where we discuss the novelty of our study.

Reviewer #2 (Recommendations for the authors):

Major issues that affect impact:

– The improvements in disease symptoms appeared to be independent of the viral load. Since AC2-26 had previously been shown to have anti-inflammatory properties these findings are, while intriguing in the context of disease, not necessarily novel.

Thank you for your comment. As discussed through the manuscript, the protective effect of AnxA1 and its mimetic peptide have been mainly demonstrated in sterile settings (e.g. inflammation triggered by LPS, uric acid crystals, DSS, and atherosclerosis genetically engineered animals), revealing various anti-inflammatory and pro-resolving actions of these molecules, including the promotion of granulocyte apoptosis, efferocytosis, and macrophage reprogramming (Fredman et al., 2015; Galvao et al., 2017; Gimenes et al., 2015; Kusters et al., 2015; Leoni et al., 2015; Locatelli et al., 2014; Sugimoto, Vago, Perretti, and Teixeira, 2019). Later, studies suggested that pro-resolving mediators can suppress inflammation and reduce the inflammatory response while improving the phagocytosis of pathogens. This was initially demonstrated for specialised lipid mediators in bacterial infection (Chiang et al., 2012; Decker et al., 2021; Sekheri et al., 2020), with subsequent studies with Annexin A1 on various infection models (Gobbetti et al., 2014; Machado et al., 2020; L. G. Oliveira et al., 2017; Tzelepis et al., 2015; Vanessa et al., 2015).

However, data on the role of resolving mediators on viral infections are scarce, with most current studies focusing on lipidic mediators of resolution (Walker et al., 2021) and insufficient data on the role of Annexin A1 in viral diseases (Schloer et al., 2019). Findings that Annexin A1 could promote influenza A virus replication in vitro were particularly surprising (Arora et al., 2016). Therefore, while studies on bacterial infection consistently revealed the ability of pro-resolving drugs to facilitate innate immune responses against the pathogen while reducing the deleterious effects of inflammation, the translational potential of these results to viral infection was less clear.

Our work is the first to access the protective role of AnxA1 in dengue infection and to reveal the therapeutic potential of Annexin A1 mimetic peptides. We show that AnxA1 can suppress the inflammatory response without hampering the protective immunity against the virus. Besides, although previous studies have suggested that AnxA1 could suppress mast cell degranulation (Gimenes et al., 2015; M. P. Oliveira et al., 2021; Parisi, Correa, and Gil, 2019; Stuqui et al., 2015), our manuscript is the first to describe the role of AnxA1 on mast cells degranulation triggered by DENV.

Finally, our findings are relevant since pathogenic immune responses are not exclusive to dengue but underlie the severity of several other viral diseases, including severe community-acquired pneumonia caused by viruses (D'Elia, Harrison, Oyston, Lukaszewski, and Clark, 2013; Perrone, Plowden, Garcia-Sastre, Katz, and Tumpey, 2008) and COVID-19 (Jose and Manuel, 2020). Therefore, other researchers could explore our data and translate it into different infectious settings, whereby targeting the AnxA1 pathway, with or without combination with antiviral drugs, holds promising therapeutic potential.

The relevance and novelty of our findings were further highlighted in the updated version of our manuscript (lines 89; 623; 648).

– It is well established that DENV does not replicate efficiently in mice, but infection can occur using mouse-adapted strains of DENV and/or immunocompromised mouse strains. Here, the authors use a combination of wt (Balb/C and C57BL/6) and mouse mutant lines (including AG129 mice which have been shown to support robust infection with mouse adapted DENV strains) and infect them with a very high dose of DENV2 isolate. There are very limited viremia data shown throughout the manuscript which makes is somewhat difficult to interpret some of the results. How much are the effects in all the non-AG129 mouse lines attributable to an acute response to the high inoculum (and no/very limited replication) vs actual infection?

Despite the limitations of modelling dengue disease in immunocompetent animals, in our study, AnxA1 and FPR2/ALX knockout animals (and their respective littermates BALB/c and C57BL/6 mice) were used as a genetic tool to investigate the role of endogenous AnxA1 in dengue disease. As mentioned by the Reviewer, DENV low passage clinical isolates do not replicate efficiently in immunocompetent mice, and for that reason, the inoculum must be higher than when using adapted strains to induce a similar disease profile. Of note, here we used 1x10⁶ PFU of DENV-2 per mouse (for immunocompetent strains): this is the same virus strain and inoculum used in recent publications (Rathore et al., 2019; St John, Rathore, Raghavan, Ng, and Abraham, 2013). Although viremia is not detectable in immunocompetent mice (St John et al., 2013), we noticed transient viral RNA detection in the spleen starting 24h after infection, peaking at 72h, followed by disappearance at 96h (Figure 2_supplement 1 and Figure 3_supplement 1). These data are in line with those presented in previous studies using the same model (St John et al., 2013; Syenina, Jagaraj, Aman, Sridharan, and St John, 2015).

Although resistant to DENV infection (Shresta et al., 2004; Yauch and Shresta, 2008), it has been reported that infection of immunocompetent mice with DENV can induce a self-limited disease, like dengue fever. While limited by the lack of detectable viremia, these models allow the study of critical immune components, such as mast cells (Rathore et al., 2019; St John et al., 2013; Syenina et al., 2015), and their contribution to the classical haematological abnormalities and altered vascular permeability observed during DENV infection (Assuncao-Miranda et al., 2010; Chen, Hofman, Kung, Lin, and Wu-Hsieh, 2007).

To complement our studies with Balb/c and C57Bl/6 mice, we have performed experiments in A129 mice since they are known to be highly susceptible to flaviviruses infections (de Freitas et al., 2019; Del Sarto et al., 2020) and develop a more severe disease manifestation that emulates several features of severe dengue in humans (Zandi et al., 2019). This model is considered a gold standard viremia model to test potential antiviral therapies (Meyts and Casanova, 2021; Williams, Zompi, Beatty, and Harris, 2009). Using this model, we aimed to evaluate if the treatment with peptide Ac_2-26 impacts viral loads, besides the alterations on disease and inflammatory parameters observed in immunocompetent animals. We have successfully recovered viable virus from samples using the plaque assay technique, as described in the Method section of the manuscript. Since A129 animals are more susceptible to infection, we could use a lower inoculum of the same non-adapted strain in this model (10³ PFU).

Therefore, we have applied two different techniques (quantification of viral RNA for immunocompetent mice and quantification of viable particles for A129 KO mice) and two different models (high and low DENV inoculums) to investigate the impact of endogenous and exogenous AnxA1 on the host ability to deal with the infection.

We have included a new paragraph in the revised manuscript discussing this important point (lines 586-605).

– In Figure 4, when using IFNα/βR-/- mice (AG129), IL-6 and CCL5 instead of MCPT-1 and CCL2 were used as markers of systemic inflammatory mediators, which is not consistent with the previous experiments done in immunocompetent mice. No results of MCPT-1 and CCL2 changes in AG129 mice was presented and the authors didn't explain why they changed the markers in this manuscript. Since MCPT-1 is a mast cell-specific product, if it is not decreased in AG129 mice treated with Ac2-26 after DENV infection, how would the authors explain the protective effects of Ac2-26 in AG129 mice in the context of DENV infection?

Thank you for this important comment. The inflammatory markers investigated in the model of severe dengue (A129 animals) were selected based on previous publications showing CCL5 and IL6 as classical inflammatory markers in dengue and a requirement of CCR5 for DENV replication and infection development (Guabiraba et al., 2010; Marques et al., 2015). Besides, liver injury, a feature of dengue disease observed in our A129 model (Binh, Matheus, Huong, Deparis, and Marechal, 2009; Trung et al., 2010) seems to correlate with high levels of CCL5 (Conceicao et al., 2010; Suksanpaisan, Cabrera-Hernandez, and Smith, 2007),

As suggested by the Reviewer, we have included data on the systemic levels of MCPT-1 and CCL2 in A129 mice subjected to the model of severe dengue (Figure 4H-J; line 383). As observed in immunocompetent mice, Ac_2-26 attenuated the systemic release of the mast cell chymase MCPT-1 induced by DENV-2 (Figure 4H), although it did not affect systemic levels of CCL2 (Figure 4I). The lack of effect of Ac_2-26 in CCL2 levels might be justified by the higher severity of the model in A129 animals, resulting in significantly higher production of CCL2 compared to immunocompetent animals infected with DENV (mean ± SD CCL2 production at the peak of 585.0 ± 61.9 for C57BL/6; 621.2 ± 100.5 for BALB/c; and 977.8 ± 110.7 for A129 mice).

– The authors showed AnxA1 memetic peptide Ac2-26 could suppress MC degranulation. However, the same experiments should also be done in parallel in AnxA1-/- and FPR2/ALX-/- mice and in BMMCs derived from them. If Ac2-26 can prevent MC degranulation, it is expected to see decreased degranulation in AnxA1 mice but not FPR2/ALX KO mice when infected with DENV, both in vivo and in BMMCs.

Thank you for your suggestion. We have included new data in the revised version of the manuscript showing the dependence of FPR2/ALX for the modulatory effect of Ac_2-26 in mast cell degranulation (Figure 6B,C; methods lines 201-207; results lines 461-469).

As predicted by the Reviewer, we verified that systemic administration of Ac_2-26 successfully reduced mast cell degranulation in AnxA1 KO mice (Figure 6B), but failed to rescue DENV-2 induced degranulation in FPR2/ALX KO animals (Figure 6C). Notably, the absence of FPR2/ALX significantly increased mast cell degranulation in response to the local challenge with DENV compared to the WT counterparts (Figure 6C).

References

Adel, F. W., Rikhi, A., Wan, S. H., Iyer, S. R., Chakraborty, H., McNulty, S.,... Chen, H. H. (2020). Annexin A1 is a Potential Novel Biomarker of Congestion in Acute Heart Failure. J Card Fail, 26(8), 727-732. doi:10.1016/j.cardfail.2020.05.012

Arora, S., Lim, W., Bist, P., Perumalsamy, R., Lukman, H. M., Li, F.,... Lim, L. H. (2016). Influenza A virus enhances its propagation through the modulation of Annexin-A1 dependent endosomal trafficking and apoptosis. Cell Death Differ, 23(7), 1243-1256. doi:10.1038/cdd.2016.19

Assuncao-Miranda, I., Amaral, F. A., Bozza, F. A., Fagundes, C. T., Sousa, L. P., Souza, D. G.,... Bozza, M. T. (2010). Contribution of macrophage migration inhibitory factor to the pathogenesis of dengue virus infection. FASEB J, 24(1), 218-228. doi:10.1096/fj.09-139469

Bandara, S. M. R., and Herath, H. (2018). Effectiveness of corticosteroid in the treatment of dengue – A systemic review. Heliyon, 4(9), e00816. doi:10.1016/j.heliyon.2018.e00816

Bandara, S. M. R., and Herath, H. M. M. T. B. (2020). Corticosteroid actions on dengue immune pathology; A review article. Clinical Epidemiology and Global Health, 8(2), 486-494. doi:10.1016/j.cegh.2019.11.001

Binh, P. T., Matheus, S., Huong, V. T., Deparis, X., and Marechal, V. (2009). Early clinical and biological features of severe clinical manifestations of dengue in Vietnamese adults. J Clin Virol, 45(4), 276-280. doi:10.1016/j.jcv.2009.04.004

Chen, H. C., Hofman, F. M., Kung, J. T., Lin, Y. D., and Wu-Hsieh, B. A. (2007). Both virus and tumor necrosis factor α are critical for endothelium damage in a mouse model of dengue virus-induced hemorrhage. J Virol, 81(11), 5518-5526. doi:10.1128/JVI.02575-06

Chiang, N., Fredman, G., Backhed, F., Oh, S. F., Vickery, T., Schmidt, B. A., and Serhan, C. N. (2012). Infection regulates pro-resolving mediators that lower antibiotic requirements. Nature, 484(7395), 524-528. doi:10.1038/nature11042

Conceicao, T. M., El-Bacha, T., Villas-Boas, C. S., Coello, G., Ramirez, J., Montero-Lomeli, M., and Da Poian, A. T. (2010). Gene expression analysis during dengue virus infection in HepG2 cells reveals virus control of innate immune response. J Infect, 60(1), 65-75. doi:10.1016/j.jinf.2009.10.003

D'Elia, R. V., Harrison, K., Oyston, P. C., Lukaszewski, R. A., and Clark, G. C. (2013). Targeting the "cytokine storm" for therapeutic benefit. Clin Vaccine Immunol, 20(3), 319-327. doi:10.1128/CVI.00636-12

Damazo, A. S., Yona, S., Flower, R. J., Perretti, M., and Oliani, S. M. (2006). Spatial and temporal profiles for anti-inflammatory gene expression in leukocytes during a resolving model of peritonitis. J Immunol, 176(7), 4410-4418. doi:10.4049/jimmunol.176.7.4410

De Caterina, R., Sicari, R., Giannessi, D., Paggiaro, P. L., Paoletti, P., Lazzerini, G.,... Parente, L. (1993). Macrophage-specific eicosanoid synthesis inhibition and lipocortin-1 induction by glucocorticoids. J Appl Physiol (1985), 75(6), 2368-2375. doi:10.1152/jappl.1993.75.6.2368

de Freitas, C. S., Higa, L. M., Sacramento, C. Q., Ferreira, A. C., Reis, P. A., Delvecchio, R.,... Souza, T. M. L. (2019). Yellow fever virus is susceptible to sofosbuvir both in vitro and in vivo. PLoS Negl Trop Dis, 13(1), e0007072. doi:10.1371/journal.pntd.0007072

Decker, C., Sadhu, S., and Fredman, G. (2021). Pro-Resolving Ligands Orchestrate Phagocytosis. Front Immunol, 12, 660865. doi:10.3389/fimmu.2021.660865

Del Sarto, J. L., de Paiva Froes Rocha, R., Bassit, L., Olmo, I. G., Valiate, B., Queiroz-Junior, C. M.,... Teixeira, M. M. (2020). 7-Deaza-7-Fluoro-2'-C-Methyladenosine Inhibits Zika virus Infection and Viral-induced Neuroinflammation. Antiviral Res, 104855. doi:10.1016/j.antiviral.2020.104855

Fredman, G., Kamaly, N., Spolitu, S., Milton, J., Ghorpade, D., Chiasson, R.,... Tabas, I. (2015). Targeted nanoparticles containing the proresolving peptide Ac2-26 protect against advanced atherosclerosis in hypercholesterolemic mice. Sci Transl Med, 7(275), 275ra220. doi:10.1126/scitranslmed.aaa1065

Galvao, I., Vago, J. P., Barroso, L. C., Tavares, L. P., Queiroz-Junior, C. M., Costa, V. V.,... Teixeira, M. M. (2017). Annexin A1 promotes timely resolution of inflammation in murine gout. Eur J Immunol, 47(3), 585-596. doi:10.1002/eji.201646551

Gimenes, A. D., Andrade, T. R., Mello, C. B., Ramos, L., Gil, C. D., and Oliani, S. M. (2015). Beneficial effect of annexin A1 in a model of experimental allergic conjunctivitis. Exp Eye Res, 134, 24-32. doi:10.1016/j.exer.2015.03.013

Gobbetti, T., Coldewey, S. M., Chen, J., McArthur, S., le Faouder, P., Cenac, N.,... Perretti, M. (2014). Nonredundant protective properties of FPR2/ALX in polymicrobial murine sepsis. Proc Natl Acad Sci U S A, 111(52), 18685-18690. doi:10.1073/pnas.1410938111

Guabiraba, R., Marques, R. E., Besnard, A. G., Fagundes, C. T., Souza, D. G., Ryffel, B., and Teixeira, M. M. (2010). Role of the chemokine receptors CCR1, CCR2 and CCR4 in the pathogenesis of experimental dengue infection in mice. PLoS ONE, 5(12), e15680. doi:10.1371/journal.pone.0015680

Jose, R. J., and Manuel, A. (2020). COVID-19 cytokine storm: the interplay between inflammation and coagulation. The Lancet Respiratory Medicine, 8(6), e46-e47. doi:10.1016/s2213-2600(20)30216-2

Kusters, D. H., Chatrou, M. L., Willems, B. A., De Saint-Hubert, M., Bauwens, M., van der Vorst, E.,... Reutelingsperger, C. P. (2015). Pharmacological Treatment with Annexin A1 Reduces Atherosclerotic Plaque Burden in LDLR-/- Mice on Western Type Diet. PLoS ONE, 10(6), e0130484. doi:10.1371/journal.pone.0130484

Leoni, G., Neumann, P. A., Kamaly, N., Quiros, M., Nishio, H., Jones, H. R.,... Nusrat, A. (2015). Annexin A1-containing extracellular vesicles and polymeric nanoparticles promote epithelial wound repair. J Clin Invest, 125(3), 1215-1227. doi:10.1172/JCI76693

Locatelli, I., Sutti, S., Jindal, A., Vacchiano, M., Bozzola, C., Reutelingsperger, C.,... Perretti, M. (2014). Endogenous annexin A1 is a novel protective determinant in nonalcoholic steatohepatitis in mice. Hepatology, 60(2), 531-544. doi:10.1002/hep.27141

Machado, M. G., Tavares, L. P., Souza, G. V. S., Queiroz-Junior, C. M., Ascencao, F. R., Lopes, M. E.,... Sousa, L. P. (2020). The Annexin A1/FPR2 pathway controls the inflammatory response and bacterial dissemination in experimental pneumococcal pneumonia. FASEB J, 34(2), 2749-2764. doi:10.1096/fj.201902172R

Marques, R. E., Guabiraba, R., Del Sarto, J. L., Rocha, R. F., Queiroz, A. L., Cisalpino, D.,... Teixeira, M. M. (2015). Dengue virus requires the CC-chemokine receptor CCR5 for replication and infection development. Immunology, 145(4), 583-596. doi:10.1111/imm.12476

Meyts, I., and Casanova, J. L. (2021). Viral infections in humans and mice with genetic deficiencies of the type I IFN response pathway. Eur J Immunol, 51(5), 1039-1061. doi:10.1002/eji.202048793

Oliveira, L. G., Souza-Testasicca, M. C., Vago, J. P., Figueiredo, A. B., Canavaci, A. M., Perucci, L. O.,... Fernandes, A. P. (2017). Annexin A1 Is Involved in the Resolution of Inflammatory Responses during Leishmania braziliensis Infection. J Immunol, 198(8), 3227-3236. doi:10.4049/jimmunol.1602028

Oliveira, M. P., Prates, J., Gimenes, A. D., Correa, S. G., and Oliani, S. M. (2021). Annexin A1 Mimetic Peptide Ac2-26 Modulates the Function of Murine Colonic and Human Mast Cells. Front Immunol, 12, 689484. doi:10.3389/fimmu.2021.689484

PAHO. (2016). Dengue: guidelines for patient care in the Region of the Americas..

Parisi, J. D. S., Correa, M. P., and Gil, C. D. (2019). Lack of Endogenous Annexin A1 Increases Mast Cell Activation and Exacerbates Experimental Atopic Dermatitis. Cells, 8(1). doi:10.3390/cells8010051

Patel, H. B., Kornerup, K. N., Sampaio, A. L., D'Acquisto, F., Seed, M. P., Girol, A. P.,... Perretti, M. (2012). The impact of endogenous annexin A1 on glucocorticoid control of inflammatory arthritis. Ann Rheum Dis, 71(11), 1872-1880. doi:10.1136/annrheumdis-2011-201180

Perretti, M., Ahluwalia, A., Harris, J. G., Goulding, N. J., and Flower, R. J. (1993). Lipocortin-1 fragments inhibit neutrophil accumulation and neutrophil-dependent edema in the mouse. A qualitative comparison with an anti-CD11b monoclonal antibody. J Immunol, 151(8), 4306-4314.

Perretti, M., and D'Acquisto, F. (2009). Annexin A1 and glucocorticoids as effectors of the resolution of inflammation. Nat Rev Immunol, 9(1), 62-70. doi:10.1038/nri2470

Perretti, M., Leroy, X., Bland, E. J., and Montero-Melendez, T. (2015). Resolution Pharmacology: Opportunities for Therapeutic Innovation in Inflammation. Trends Pharmacol Sci, 36(11), 737-755. doi:10.1016/j.tips.2015.07.007

Perrone, L. A., Plowden, J. K., Garcia-Sastre, A., Katz, J. M., and Tumpey, T. M. (2008). H5N1 and 1918 pandemic influenza virus infection results in early and excessive infiltration of macrophages and neutrophils in the lungs of mice. PLoS Pathog, 4(8), e1000115. doi:10.1371/journal.ppat.1000115

Purvis, G. S. D., Collino, M., Loiola, R. A., Baragetti, A., Chiazza, F., Brovelli, M.,... Thiemermann, C. (2019). Identification of AnnexinA1 as an Endogenous Regulator of RhoA, and Its Role in the Pathophysiology and Experimental Therapy of Type-2 Diabetes. Front Immunol, 10, 571. doi:10.3389/fimmu.2019.00571

Rathore, A. P., Mantri, C. K., Aman, S. A., Syenina, A., Ooi, J., Jagaraj, C. J.,... St John, A. L. (2019). Dengue virus-elicited tryptase induces endothelial permeability and shock. J Clin Invest, 129(10), 4180-4193. doi:10.1172/JCI128426

Rowe, E. K., Leo, Y. S., Wong, J. G., Thein, T. L., Gan, V. C., Lee, L. K., and Lye, D. C. (2014). Challenges in dengue fever in the elderly: atypical presentation and risk of severe dengue and hospital-acquired infection [corrected]. PLoS Negl Trop Dis, 8(4), e2777. doi:10.1371/journal.pntd.0002777

Sangkaew, S., Ming, D., Boonyasiri, A., Honeyford, K., Kalayanarooj, S., Yacoub, S.,... Holmes, A. (2021). Risk predictors of progression to severe disease during the febrile phase of dengue: a systematic review and meta-analysis. The Lancet Infectious Diseases, 21(7), 1014-1026. doi:10.1016/s1473-3099(20)30601-0

Santana, B. B., Queiroz, M. A. F., Cerveira, R. A., Rodrigues, C. M., da Silva Graca Amoras, E., da Costa, C. A.,... Vallinoto, A. C. R. (2021). Low Annexin A1 level in HTLV-1 infected patients is a potential biomarker for the clinical progression and diagnosis of HAM/TSP. BMC Infect Dis, 21(1), 219. doi:10.1186/s12879-021-05917-y

Schloer, S., Hubel, N., Masemann, D., Pajonczyk, D., Brunotte, L., Ehrhardt, C.,... Rescher, U. (2019). The annexin A1/FPR2 signaling axis expands alveolar macrophages, limits viral replication, and attenuates pathogenesis in the murine influenza A virus infection model. FASEB J, fj201901265R. doi:10.1096/fj.201901265R

Sekheri, M., El Kebir, D., Edner, N., and Filep, J. G. (2020). 15-Epi-LXA4 and 17-epi-RvD1 restore TLR9-mediated impaired neutrophil phagocytosis and accelerate resolution of lung inflammation. Proc Natl Acad Sci U S A, 117(14), 7971-7980. doi:10.1073/pnas.1920193117

Senchenkova, E. Y., Ansari, J., Becker, F., Vital, S. A., Al-Yafeai, Z., Sparkenbaugh, E. M.,... Gavins, F. N. E. (2019). Novel Role for the AnxA1-Fpr2/ALX Signaling Axis as a Key Regulator of Platelet Function to Promote Resolution of Inflammation. Circulation, 140(4), 319-335. doi:10.1161/CIRCULATIONAHA.118.039345

Shresta, S., Kyle, J. L., Snider, H. M., Basavapatna, M., Beatty, P. R., and Harris, E. (2004). Interferon-dependent immunity is essential for resistance to primary dengue virus infection in mice, whereas T- and B-cell-dependent immunity are less critical. J Virol, 78(6), 2701-2710. doi:10.1128/jvi.78.6.2701-2710.2004

Solito, E., Mulla, A., Morris, J. F., Christian, H. C., Flower, R. J., and Buckingham, J. C. (2003). Dexamethasone induces rapid serine-phosphorylation and membrane translocation of annexin 1 in a human folliculostellate cell line via a novel nongenomic mechanism involving the glucocorticoid receptor, protein kinase C, phosphatidylinositol 3-kinase, and mitogen-activated protein kinase. Endocrinology, 144(4), 1164-1174. doi:10.1210/en.2002-220592

St John, A. L., Rathore, A. P., Raghavan, B., Ng, M. L., and Abraham, S. N. (2013). Contributions of mast cells and vasoactive products, leukotrienes and chymase, to dengue virus-induced vascular leakage. eLife, 2, e00481. doi:10.7554/eLife.00481

Stuqui, B., de Paula-Silva, M., Carlos, C. P., Ullah, A., Arni, R. K., Gil, C. D., and Oliani, S. M. (2015). Ac2-26 Mimetic Peptide of Annexin A1 Inhibits Local and Systemic Inflammatory Processes Induced by Bothrops moojeni Venom and the Lys-49 Phospholipase A2 in a Rat Model. PLoS ONE, 10(7), e0130803. doi:10.1371/journal.pone.0130803

Sugimoto, M. A., Vago, J. P., Perretti, M., and Teixeira, M. M. (2019). Mediators of the Resolution of the Inflammatory Response. Trends Immunol, 40(3), 212-227. doi:10.1016/j.it.2019.01.007

Suksanpaisan, L., Cabrera-Hernandez, A., and Smith, D. R. (2007). Infection of human primary hepatocytes with dengue virus serotype 2. J Med Virol, 79(3), 300-307. doi:10.1002/jmv.20798

Syenina, A., Jagaraj, C. J., Aman, S. A., Sridharan, A., and St John, A. L. (2015). Dengue vascular leakage is augmented by mast cell degranulation mediated by immunoglobulin Fcgamma receptors. eLife, 4. doi:10.7554/eLife.05291

Trung, D. T., Thao le, T. T., Hien, T. T., Hung, N. T., Vinh, N. N., Hien, P. T.,... Wills, B. (2010). Liver involvement associated with dengue infection in adults in Vietnam. Am J Trop Med Hyg, 83(4), 774-780. doi:10.4269/ajtmh.2010.10-0090

Tzelepis, F., Verway, M., Daoud, J., Gillard, J., Hassani-Ardakani, K., Dunn, J.,... Divangahi, M. (2015). Annexin1 regulates DC efferocytosis and cross-presentation during Mycobacterium tuberculosis infection. J Clin Invest, 125(2), 752-768. doi:10.1172/JCI77014

Vago, J. P., Nogueira, C. R., Tavares, L. P., Soriani, F. M., Lopes, F., Russo, R. C.,... Sousa, L. P. (2012). Annexin A1 modulates natural and glucocorticoid-induced resolution of inflammation by enhancing neutrophil apoptosis. J Leukoc Biol, 92(2), 249-258. doi:10.1189/jlb.0112008

Vanessa, K. H., Julia, M. G., Wenwei, L., Michelle, A. L., Zarina, Z. R., Lina, L. H., and Sylvie, A. (2015). Absence of Annexin A1 impairs host adaptive immunity against Mycobacterium tuberculosis in vivo. Immunobiology, 220(5), 614-623. doi:10.1016/j.imbio.2014.12.001

Walker, K. H., Krishnamoorthy, N., Bruggemann, T. R., Shay, A. E., Serhan, C. N., and Levy, B. D. (2021). Protectins PCTR1 and PD1 Reduce Viral Load and Lung Inflammation During Respiratory Syncytial Virus Infection in Mice. Front Immunol, 12, 704427. doi:10.3389/fimmu.2021.704427

WHO. (‎2009). Dengue guidelines for diagnosis, treatment, prevention and control : new edition. : World Health Organization.

Williams, K. L., Zompi, S., Beatty, P. R., and Harris, E. (2009). A mouse model for studying dengue virus pathogenesis and immune response. Ann N Y Acad Sci, 1171 Suppl 1, E12-23. doi:10.1111/j.1749-6632.2009.05057.x

Xu, X., Gao, W., Li, L., Hao, J., Yang, B., Wang, T.,... Jiao, L. (2021). Annexin A1 protects against cerebral ischemia-reperfusion injury by modulating microglia/macrophage polarisation via FPR2/ALX-dependent AMPK-mTOR pathway. J Neuroinflammation, 18(1), 119. doi:10.1186/s12974-021-02174-3

Yauch, L. E., and Shresta, S. (2008). Mouse models of dengue virus infection and disease. Antiviral Res, 80(2), 87-93. doi:10.1016/j.antiviral.2008.06.010

Zandi, K., Bassit, L., Amblard, F., Cox, B. D., Hassandarvish, P., Moghaddam, E.,... Schinazi, R. F. (2019). Nucleoside Analogs with Selective Antiviral Activity against Dengue Fever and Japanese Encephalitis Viruses. Antimicrob Agents Chemother, 63(7). doi:10.1128/AAC.00397-19

Author details

1. Vivian Vasconcelos Costa

   Department of Morphology, Institute of Biological Sciences, Universidade Federal de Minas Gerais, Belo Horizonte, Brazil

   Contribution

   Conceptualization, Data curation, Formal analysis, Funding acquisition, Investigation, Methodology, Project administration, Supervision, Validation, Visualization, Writing – original draft

   Contributed equally with

   Michelle A Sugimoto

   Competing interests

   No competing interests declared

   "This ORCID iD identifies the author of this article:" 0000-0002-0175-642X

2. Michelle A Sugimoto

   1. Department of Biochemistry and Immunology, Institute of Biological Sciences, Universidade Federal de Minas Gerais, Belo Horizonte, Brazil
   2. School of Medicine, Universidade Federal de Minas Gerais, Belo Horizonte, Brazil
   3. William Harvey Research Institute, Barts and The London School of Medicine and Dentistry, Queen Mary University of London, London, United Kingdom

   Contribution

   Conceptualization, Data curation, Formal analysis, Investigation, Methodology, Project administration, Validation, Visualization, Writing – original draft

   Contributed equally with

   Vivian Vasconcelos Costa

   Competing interests

   No competing interests declared

   "This ORCID iD identifies the author of this article:" 0000-0002-4527-6065

3. Josy Hubner

   Department of Morphology, Institute of Biological Sciences, Universidade Federal de Minas Gerais, Belo Horizonte, Brazil

   Contribution

   Investigation

   Competing interests

   No competing interests declared

4. Caio S Bonilha

   1. Department of Biochemistry and Immunology, Institute of Biological Sciences, Universidade Federal de Minas Gerais, Belo Horizonte, Brazil
   2. Institute of Infection, Immunity and Inflammation, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow, United Kingdom

   Contribution

   Visualization, Writing - review and editing

   Competing interests

   No competing interests declared

5. Celso Martins Queiroz-Junior

   Department of Morphology, Institute of Biological Sciences, Universidade Federal de Minas Gerais, Belo Horizonte, Brazil

   Contribution

   Data curation, Investigation, Visualization, Writing - review and editing

   Competing interests

   No competing interests declared

6. Marcela Helena Gonçalves-Pereira

   Department of Biochemistry and Immunology, Institute of Biological Sciences, Universidade Federal de Minas Gerais, Belo Horizonte, Brazil

   Contribution

   Data curation, Investigation, Resources, Writing - review and editing

   Competing interests

   No competing interests declared

7. Jianmin Chen

   William Harvey Research Institute, Barts and The London School of Medicine and Dentistry, Queen Mary University of London, London, United Kingdom

   Contribution

   Investigation, Writing - review and editing

   Competing interests

   No competing interests declared

8. Thomas Gobbetti

   William Harvey Research Institute, Barts and The London School of Medicine and Dentistry, Queen Mary University of London, London, United Kingdom

   Contribution

   Investigation

   Competing interests

   No competing interests declared

9. Gisele Olinto Libanio Rodrigues

   Department of Biochemistry and Immunology, Institute of Biological Sciences, Universidade Federal de Minas Gerais, Belo Horizonte, Brazil

   Contribution

   Data curation, Investigation, Methodology

   Competing interests

   No competing interests declared

10. Jordana L Bambirra

    Department of Microbiology, Institute of Biological Sciences, Universidade Federal de Minas Gerais, Belo Horizonte, Brazil

    Contribution

    Investigation, Methodology

    Competing interests

    No competing interests declared

11. Ingredy B Passos

    Department of Morphology, Institute of Biological Sciences, Universidade Federal de Minas Gerais, Belo Horizonte, Brazil

    Contribution

    Data curation, Investigation, Methodology

    Competing interests

    No competing interests declared

12. Carla Elizabeth Machado Lopes

    Department of Morphology, Institute of Biological Sciences, Universidade Federal de Minas Gerais, Belo Horizonte, Brazil

    Contribution

    Investigation

    Competing interests

    No competing interests declared

13. Thaiane P Moreira

    Department of Microbiology, Institute of Biological Sciences, Universidade Federal de Minas Gerais, Belo Horizonte, Brazil

    Contribution

    Investigation

    Competing interests

    No competing interests declared

14. Kennedy Bonjour

    Department of Biology, Institute of Biological Sciences, Federal University of Juiz de Fora, Juiz de Fora, Brazil

    Contribution

    Data curation, Formal analysis, Investigation, Methodology, Validation, Writing - review and editing

    Competing interests

    No competing interests declared

15. Rossana CN Melo

    Department of Biology, Institute of Biological Sciences, Federal University of Juiz de Fora, Juiz de Fora, Brazil

    Contribution

    Data curation, Investigation, Methodology

    Competing interests

    No competing interests declared

16. Milton AP Oliveira

    Tropical Pathology and Public Health Institute, Universidade Federal de Goiás, Goiânia, Brazil

    Contribution

    Resources

    Competing interests

    No competing interests declared

17. Marcus Vinicius M Andrade

    School of Medicine, Universidade Federal de Minas Gerais, Belo Horizonte, Brazil

    Contribution

    Methodology, Resources, Supervision

    Competing interests

    No competing interests declared

18. Lirlândia Pires Sousa

    Department of Clinical and Toxicological Analyses, School of Pharmacy, Universidade Federal de Minas Gerais, Belo Horizonte, Brazil

    Contribution

    Investigation, Methodology, Resources

    Competing interests

    No competing interests declared

    "This ORCID iD identifies the author of this article:" 0000-0002-1042-9762

19. Danielle Gloria Souza

    Department of Microbiology, Institute of Biological Sciences, Universidade Federal de Minas Gerais, Belo Horizonte, Brazil

    Contribution

    Conceptualization, Investigation, Methodology, Supervision, Visualization

    Competing interests

    No competing interests declared

20. Helton da Costa Santiago

    Department of Biochemistry and Immunology, Institute of Biological Sciences, Universidade Federal de Minas Gerais, Belo Horizonte, Brazil

    Contribution

    Conceptualization, Data curation, Funding acquisition, Project administration, Resources, Supervision, Visualization

    Competing interests

    No competing interests declared

21. Mauro Perretti

    1. William Harvey Research Institute, Barts and The London School of Medicine and Dentistry, Queen Mary University of London, London, United Kingdom
    2. Centre for Inflammation and Therapeutic Innovation, Queen Mary University of London, London, United Kingdom

    Contribution

    Conceptualization, Funding acquisition, Methodology, Project administration, Resources, Supervision, Visualization, Writing – original draft

    Contributed equally with

    Mauro Martins Teixeira

    For correspondence

    m.perretti@qmul.ac.uk

    Competing interests

    is on the Scientific Advisory Board of ResoTher Pharma AS, which is interested in developing Annexin A1-derived peptides for cardiovascular settings

    "This ORCID iD identifies the author of this article:" 0000-0003-2068-3331

22. Mauro Martins Teixeira

    Department of Biochemistry and Immunology, Institute of Biological Sciences, Universidade Federal de Minas Gerais, Belo Horizonte, Brazil

    Contribution

    Conceptualization, Funding acquisition, Methodology, Project administration, Resources, Supervision, Visualization, Writing – original draft

    Contributed equally with

    Mauro Perretti

    For correspondence

    mmtex.ufmg@gmail.com

    Competing interests

    No competing interests declared

    "This ORCID iD identifies the author of this article:" 0000-0002-6944-3008

• 1,257

  Page views

• 170

  Downloads

• 4

  Citations

Article citation count generated by polling the highest count across the following sources: Crossref, PubMed Central, Scopus.