Interferon-gamma signaling is downregulated in peripheral blood mononuclear cells from asthma patients
A sinalização de interferon-gama é diminuída em células mononucleares do sangue periférico de pacientes asmáticos
Carolina Valadares Nunes; Laila Rigolin Fortunato; Jhony Robison Oliveira; Aline Beatriz Mahler Pereira; Gabriela Rister Figueiredo Irie; Sarah Cristina Sato Vaz Tanaka; Marcos Vinícius Silva; Wilson Carneiro Silva-Jr; Marly Aparecida Spadotto Balarin; Paulo Roberto Silva; David Nascimento Silva Teixeira; Alexandre Paula Rogerio
Abstract
Introduction: Interferon-gamma (IFN-g) signaling is mediated by crosstalk of receptors, such as IFN-g receptor 1 (IFN-g R1), transcription factors, such as signal transducer and activator of transcription 1 (STAT1) and suppressors of cytokine signaling 1 (SOCS1). Here, we evaluated the role of IFN-g signaling in peripheral blood mononuclear cells (PBMCs) from asthma patients and control individuals. Methods: PBMCs from adult healthy nonasthmatic controls (n = 12; male and female, 18-60 years old) and patients diagnosed with asthma (n = 18; male and female, 18-60 years old) were stimulated with IFN-g (0.25, 0.5 and/or 1.0 ng/mL) and, after 24h, the production of CXC motif chemokine 10 (CXCL10) was evaluated (by enzyme linked immunosorbent assay) as well as the expression of IFN-g R1, STAT1 (both by flow cytometry assay) and SOCS1 (by real-time qPCR assay). Results: CXCL10 production was reduced in a dose-dependent manner in PBMCs from asthma patients stimulated with IFN-g when compared to control individuals. While IFN-g induced an increase in IFN-g R1 expression and phosphorylated STAT1 (pSTAT1) activation in PBMCs from the control group, a reduction in both IFN-g R1 and pSTAT1 was observed in PBMCs from asthma patients. IFN-g increased SOCS1 mRNA expression in PBMCs from asthma patients when compared to IFN-g-stimulated cells from control individuals. Conclusion: Taken together, our results demonstrated that IFN-g signaling is downregulated in asthma patients.
Keywords
Resumo
Introdução: A sinalização de interferon-gama (IFN-g) é mediada por receptores, como o receptor 1 de IFN-gama (IFN-gR1), fatores de transcrição, como o transdutor de sinal e o ativador de transcrição 1 (STAT1) e supressores de sinalização de citocina 1 (SOCS1). Neste trabalho, avaliamos o papel da sinalização de IFN-g em células mononucleares do sangue periférico (PBMCs) de indivíduos com asma e controle. Métodos: Células mononucleares do sangue periférico (PBMCs) de adultos saudáveis e não asmáticos (n = 12, homens e mulheres, 18-60 anos) e pacientes diagnosticados com asma (n = 18, homens e mulheres, 18-60 anos) foram estimuladas com IFN-g (0,25, 0,5 e/ou 1,0 ng/mL) e após 24 horas a produção de CXCL10 foi avaliada por ensaio de imunoabsorção enzimática (ELISA), bem como o receptor 1 de IFN-g (IFN-g R1). Também foram avaliadas as expressões do transdutor de sinal e ativador da transcrição 1 (STAT1) (por citometria de fluxo) e supressor de expressão de sinalização de citocinas 1 (SOCS1) (por ensaio qPCR em tempo real). Resultados: A produção de CXCL10, uma quimiocina induzida por IFNg, foi reduzida de maneira dependente da dose em PBMCs de pacientes com asma estimulados com IFN-g (0,25-1,0 ng/mL) quando comparado ao grupo controle. Enquanto IFN-g induziu um aumento da expressão de IFN-g R1 e ativação da fosforilação de STAT1 (pSTAT1) em PBMCs do grupo controle, uma redução de ambas (IFN-g R1 e pSTAT1) foi observada em PBMCs de pacientes com asma. O IFN-g aumentou as PBMCs de expressão do mRNA de SOCS1 de pacientes com asma quando comparado às células estimuladas por IFN-g do controle. Conclusão: Em conjunto, nossos resultados demonstraram que a sinalização de IFN-g é sub-regulada em pacientes com asma.
Palavras-chave
References
1. Fanta CH. Asthma. N Engl J Med. 2009;360:1002-14. doi: 10.1056/ NEJMra0804579.
2. Zhu J, Message SD, Mallia P, Kebadze T, Contoli M, Ward CK, et al. Bronchial mucosal IFN-α/β and pattern recognition receptor expression in patients with experimental rhinovirus-induced asthma exacerbations. J Allergy Clin Immunol. 2019;143(1):114-25.e4. doi:10.1016/j.jaci.2018.04.003.
3. Xiaoyu H, Lionel B, Ivashkiv I. Cross-regulation of Signaling and Immune Responses by IFN-g and STAT1. Immunity. 2009; 31:539‑50. doi: 10.1016/j.immuni.2009.09.002.
4. Deo SS, Mistry KJ, Kakade AM, Niphadkar PV. Role played by Th2 type cytokines in IgE mediated allergy and asthma. Lung India. 2010 Apr;27(2):66-71. doi: 10.4103/0970-2113.63609. PMID: 20616938; PMCID: PMC2893428.
5. Figueiredo CA, Rodrigues LC, Alcantara-Neves NM, Cooper PJ, Amorim LD, Silva NB, et al. Does IFN-g play a role on the pathogenesis of non-atopic asthma in Latin America children? Allergy Asthma Clin Immunol. 2012 Dec 19;8(1):18. doi: 10.1186/1710-1492-8-18. PMID: 23253516; PMCID: PMC3564733.
6. Raundhal M, Morse C, Khare A, Oriss TB, Milosevic J, Trudeau J, et al. High IFN-g and low SLPI mark severe asthma in mice and humans. J Clin Invest. 2015;125(8):3037-50. doi:10.1172/JCI80911.
7. Ying S, Durham SR, Corrigan CJ, Hamid Q, Kay AB. Phenotype of cells expressing mRNA for TH2-type (interleukin 4 and interleukin 5) and TH1-type (interleukin 2 and interferon gamma) cytokines in bronchoalveolar lavage and bronchial biopsies from atopic asthmatic and normal control subjects. Am J Respir Cell Mol Biol. 1995;12:477-87.
8. Piehler J, Thomas C, Garcia kC, Schreiber G. Structural and dynamic determinants of type I interferon receptor asSDbly and their functional interpretation. Immunological reviews. 2012;250:317-4. doi: 10.1111/imr.12001.
9. Ivashkiv LB, Donlin LT. Regulation of type I interferon responses. Nature reviews Immunology. 2014;14:36-49. doi:10.1038/nri3581.
10. Yasukawa H, Sasaki A, Yoshimura A. Negative regulation of cytokine signaling pathways. Annu Rev Immunol. 2000;18:143-64. doi: 10.1146/annurev.immunol.18.1.143.
11. Wenzel SE. Asthma: defining of the persistent adult phenotypes. Lancet. 2006;386:804-13. doi:10.1016/S0140-6736(06)69290-8.
12. Zambalde ÉP, Teixeira MM, Favarin DC, de Oliveira JR, Magalhães ML, Cunha MM, et al. The anti-inflammatory and pro-resolution effects of aspirin-triggered RvD1 (AT-RvD1) on peripheral blood mononuclear cells from patients with severe asthma. Int Immunopharmacol. 2016 Jun;35:142-8. doi: 10.1016/j. intimp.2016.03.014.
13. de Oliveira JR, Favarin DC, Tanaka SC, Balarin MA, Teixeira DN, Levy BD, et al. AT-RvD1 modulates CCL-2 and CXCL-8 production and NF-κB, STAT-6, SOCS1, and SOCS3 expression on bronchial epithelial cells stimulated with IL-4. Biomed Res Int. 2015;15:1-8. doi: 10.1155/2015/178369.
14. Lee HJ, Choi BH, Min BH, Park SR. Changes in surface markers of human mesenchymal stem cells during the chondrogenic differentiation and dedifferentiation processes in vitro. Arthritis Rheum. 2009;60:2325-32. doi: 10.1002/art.24786.
15. Kumar RK, Webb DC, Herbert C, Foster PS. Interferon-gamma as a possible target in chronic asthma. Inflamm Allergy Drug Targets. 2006;4:253-9. doi: 10.2174/187152806779010909.
16. Medoff BD, Sauty A, Tager AM, Maclean JA, Smith RN, Mathew A, Dufour JH, Luster AD. IFN-gamma-inducible protein 10 (CXCL10) contributes to airway hyperreactivity and airway inflammation in a mouse model of asthma. J Immunol. 2002;168:5278-86. doi: https://doi.org/10.4049/jimmunol.168.10.5278.
17. Lin Y, Yan H, Xiao Y, Piao H, Xiang R, Jian L, et al. Attenuation of antigen-induced airway hyperresponsiveness and inflammation in CXCR3 knockout mice. Respir Res. 2011;12:123-31. doi:10.1186/1465-9921-12-123.
18. Cohn L, Homer RJ, Niu N, Bottomly K. T helper 1 cells and interferon gamma regulate allergic airway inflammation and mucus production. J Exp Med. 1999;190:1309-18. doi: 10.1084/jem.190.9.1309.
19. Pinto LA. STAT1 gene variations, IgE regulation and atopy. Allergy. 2007;62:1456-61. doi:10.1111/j.1398-9995.2007.01479.
20. Nakao F. Association of IFN-g and IFN regulatory factor 1 polymorphisms with childhood atopic asthma. J Allergy Clin Immunol.2001;107:499-504. doi:10.1067/mai.2001.113051.
21. Sampath D, Castro M, Look DC, Holtzman MJ. Constitutive activation of an epithelial signal transducer and activator of transcription (STAT) pathway in asthma. J Clin Invest. 1999;103:1353-61. doi:10.1172/ JCI6130.
22. Cianciulli A, Calvello R, Porro C, Trotta T, Panaro MA. Understanding the role of SOCS signaling in neurodegenerative diseases: Current and emerging concepts. Cytokine Growth Factor Rev. 2017;37:67‑79. doi:10.1016/j.cytogfr.2017.07.005.
23. Rauch I, Müller M, Decker T. The regulation of inflammation by interferons and their STATs. JAKSTAT. 2013 Jan 1;2(1):e23820. doi:10.4161/jkst.23820. PMID: 24058799; PMCID: PMC3670275.
24. Alexander WS, Starr R, Fenner JE, Scott CL, Handman E, Sprigg NS, et al. SOCS1 is a critical inhibitor of interferon gamma signaling and prevents the potentially fatal neonatal actions of this cytokine. Cell. 1999;5:597-608. doi:https://doi.org/10.1016/S0092-8674(00)80047-1.
25. Mostecki J, Cassel SL, Klimecki WT, Stern DA, Knisz J, Iwashita S, et al. A SOCS-1 promoter variant is associated with total serum IgE levels. J Immunol. 2011 Sep 1;187(5):2794-802. doi: 10.4049/ jimmunol.0902569. Epub 2011 Jul 27. PMID: 21795592; PMCID: PMC3159751.
26. Harada M, Nakashima K, Hirota T, Shimizu M, Doi S, Fujita K, et al. Functional polymorphism in the suppressor of cytokine signaling 1 gene associated with adult asthma. Am J Respir Cell Mol Biol. 2007;36(4):491-6. doi:10.1165/rcmb.2006-0090OC.
Submitted date:
05/14/2020
Accepted date:
05/30/2020
Facebook
Google+
Twitter
LinkedIn
Mendeley
StumbleUpon
CiteULike
Reddit
Email