Respiratory syncytial virus increases eosinophil extracellular traps in a murine model of asthma - PubMed (original) (raw)
Respiratory syncytial virus increases eosinophil extracellular traps in a murine model of asthma
Josiane Silva Silveira et al. Asia Pac Allergy. 2019.
Abstract
Background: Respiratory viral infections are the leading cause of asthma exacerbations. Eosinophil activation results in the formation of eosinophil extracellular traps (EETs), which release web-like structures of DNA and proteins that bind, disarm and extracellularly kill pathogens.
Objective: We investigated whether the respiratory syncytial virus (RSV) in vitro could induce EETs in bronchoalveolar lavage fluid eosinophils in a murine model of asthma.
Methods: BALB/cJ mice (6-8 weeks old) were sensitized with 2 subcutaneous injections of ovalbumin (20 μg) on days 0 and 7, followed by three intranasal challenges with ovalbumin (100 μg) on days 14, 15, and 16 of the protocol. The control group received Dulbecco's phosphate-buffered saline. Bronchoalveolar lavage fluid eosinophils of ovalbumin group or control group were stimulated with RSV (103 PFU/mL) in vitro for 3 hours. After that, culture supernatant was collected to perform the analyses proposed in this study.
Results: We verified an increase in extracellular DNA concentration in bronchoalveolar lavage fluid eosinophils from ovalbumin group stimulated with RSV (103 PFU/mL) in vitro, which was confirmed by confocal microscopy. We demonstrated that most cells are negative for annexin V and propidium iodide in all groups evaluated. Also, RSV in vitro decreased interferon-ɣ in culture supernatant when compared to the ovalbumin group.
Conclusion: In this study, we demonstrated for the first time that RSV in vitro induces EETs formation in eosinophils from asthmatic mice.
Keywords: Asthma; Eosinophil peroxidase; Extracellular traps; Inflammation; Respiratory syncytial viruses.
Copyright © 2019. Asia Pacific Association of Allergy, Asthma and Clinical Immunology.
Conflict of interest statement
Conflict of Interest: The authors have no financial conflicts of interest.
Figures
Fig. 1. Ovalbumin-induced airway eosinophilic inflammation. (A) Total cell counts in BALF. (B-E) Differential cell counts in BALF (eosinophils, macrophages, neutrophils, and lymphocytes. (F) Representative illustration of BALF cells stained with H&E (×400, arrows indicate eosinophils). Results are expressed as mean ± standard deviation, for 7–10 animals in each group, of 3 independent experiments, **p < 0.01, ***p < 0.001. BALF, bronchoalveolar lavage fluid; OVA, ovalbumin.
Fig. 2. RSV in vitro induces EETs colocalized with EPO in BALF cells from asthmatic mice. (A) Effect of different concentrations of RSV (103–106 PFU/mL) in vitro in extracellular DNA concentration. (B) Extracellular DNA concentration in BALF cells from asthmatic and control mice stimulated with RSV (103 PFU/mL) in vitro or unstimulated. (C) EETs release in BALF cells from asthmatic and control mice stimulated with RSV (103 PFU/mL) in vitro or unstimulated (×630, arrows indicate EETs formation). (D) Analysis of annexin V binding and PI uptake in BALF cells from all groups. Results are expressed as mean ± standard deviation, for 7–10 animals in each group, of 3 independent experiments, *p < 0.05, **p < 0.01, ***p < 0.001. BALF, bronchoalveolar lavage fluid; DPBS, Dulbecco's phosphate-buffered saline; EPO, eosinophil peroxidase; OVA, ovalbumin; PFU, plaque-forming unit; PI, propidium iodide; RSV, respiratory syncytial virus; FITC, fluorescein isothiocyanate.
Fig. 3. Effect of RSV in BALF cells from asthmatic mice in cytokines levels and EPO activity. (A) IL-4, (B) IFN-ɣ, and (C) EPO activity in from BALF cells (2 × 105 /mL) from asthmatic mice stimulated with RSV (103 PFU/mL) in vitro or unstimulated. Results are expressed as mean ± standard deviation, for 5 animals in each group, of 3 independent experiments, **p < 0.01. BALF, bronchoalveolar lavage fluid; EPO, eosinophil peroxidase; DPBS, Dulbecco's phosphate-buffered saline; IL-4, interleukin-4; IFN-ɣ, interferon-ɣ; OVA, ovalbumin; RSV, respiratory syncytial virus.
Similar articles
- Eosinophil extracellular traps in asthma: implications for pathogenesis and therapy.
Shen K, Zhang M, Zhao R, Li Y, Li C, Hou X, Sun B, Liu B, Xiang M, Lin J. Shen K, et al. Respir Res. 2023 Sep 26;24(1):231. doi: 10.1186/s12931-023-02504-4. Respir Res. 2023. PMID: 37752512 Free PMC article. Review. - Extracellular DNA traps in bronchoalveolar fluid from a murine eosinophilic pulmonary response.
Cunha AA, Porto BN, Nuñez NK, Souza RG, Vargas MH, Silveira JS, Souza TT, Jaeger N, Pitrez PM. Cunha AA, et al. Allergy. 2014 Dec;69(12):1696-700. doi: 10.1111/all.12507. Epub 2014 Sep 13. Allergy. 2014. PMID: 25130372 - Respiratory syncytial virus reverses airway hyperresponsiveness to methacholine in ovalbumin-sensitized mice.
Aeffner F, Davis IC. Aeffner F, et al. PLoS One. 2012;7(10):e46660. doi: 10.1371/journal.pone.0046660. Epub 2012 Oct 2. PLoS One. 2012. PMID: 23056391 Free PMC article. - [Combined effects of neonatal Bacillus Calmette-Guerin vaccination and respiratory syncytial infection on experimental asthma in mice].
Li R, Liu EM, Yang XQ, Wang LJ. Li R, et al. Zhonghua Er Ke Za Zhi. 2006 Jun;44(6):420-4. Zhonghua Er Ke Za Zhi. 2006. PMID: 16836848 Chinese. - Eosinophil extracellular vesicles and DNA traps in allergic inflammation.
Weihrauch T, Melo RCN, Gray N, Voehringer D, Weller PF, Raap U. Weihrauch T, et al. Front Allergy. 2024 Aug 1;5:1448007. doi: 10.3389/falgy.2024.1448007. eCollection 2024. Front Allergy. 2024. PMID: 39148911 Free PMC article. Review.
Cited by
- The role of extracellular traps released by neutrophils, eosinophils, and macrophages in asthma.
Gu W, Huang C, Chen G, Kong W, Zhao L, Jie H, Zhen G. Gu W, et al. Respir Res. 2024 Jul 30;25(1):290. doi: 10.1186/s12931-024-02923-x. Respir Res. 2024. PMID: 39080638 Free PMC article. Review. - Eosinophils as drivers of bacterial immunomodulation and persistence.
Parrish KM, Gestal MC. Parrish KM, et al. Infect Immun. 2024 Sep 10;92(9):e0017524. doi: 10.1128/iai.00175-24. Epub 2024 Jul 15. Infect Immun. 2024. PMID: 39007622 Free PMC article. Review. - Ally, adversary, or arbitrator? The context-dependent role of eosinophils in vaccination for respiratory viruses and subsequent breakthrough infections.
Chang LA, Schotsaert M. Chang LA, et al. J Leukoc Biol. 2024 Jul 25;116(2):224-243. doi: 10.1093/jleuko/qiae010. J Leukoc Biol. 2024. PMID: 38289826 Free PMC article. Review. - Eosinophil extracellular traps in asthma: implications for pathogenesis and therapy.
Shen K, Zhang M, Zhao R, Li Y, Li C, Hou X, Sun B, Liu B, Xiang M, Lin J. Shen K, et al. Respir Res. 2023 Sep 26;24(1):231. doi: 10.1186/s12931-023-02504-4. Respir Res. 2023. PMID: 37752512 Free PMC article. Review. - The "Self-Sacrifice" of ImmuneCells in Sepsis.
Wen X, Xie B, Yuan S, Zhang J. Wen X, et al. Front Immunol. 2022 Apr 29;13:833479. doi: 10.3389/fimmu.2022.833479. eCollection 2022. Front Immunol. 2022. PMID: 35572571 Free PMC article. Review.
References
- Psarras S, Papadopoulos NG, Johnston SL. Pathogenesis of respiratory syncytial virus bronchiolitis-related wheezing. Paediatr Respir Rev. 2004;5(Suppl A):S179–S184. - PubMed
- Yousefi S, Gold JA, Andina N, Lee JJ, Kelly AM, Kozlowski E, Schmid I, Straumann A, Reichenbach J, Gleich GJ, Simon HU. Catapult-like release of mitochondrial DNA by eosinophils contributes to antibacterial defense. Nat Med. 2008;14:949–953. - PubMed
- Cunha AA, Porto BN, Nuñez NK, Souza RG, Vargas MH, Silveira JS, Souza TT, Jaeger N, Pitrez PM. Extracellular DNA traps in bronchoalveolar fluid from a murine eosinophilic pulmonary response. Allergy. 2014;69:1696–1700. - PubMed
- da Cunha AA, Nuñez NK, de Souza RG, Moraes Vargas MH, Silveira JS, Antunes GL, Durante L da S, Porto BN, Marczak ES, Jones MH, Pitrez PM. Recombinant human deoxyribonuclease therapy improves airway resistance and reduces DNA extracellular traps in a murine acute asthma model. Exp Lung Res. 2016;42:66–74. - PubMed
LinkOut - more resources
Full Text Sources