EXPRESSION OF STATS PROTEINS IN THE DIFFERENTIATION OF NAIVE TCD4+ CELLS IN AN EXPERIMENTAL MODEL OF OVERLAPPING ASTHMA AND CHRONIC OBSTRUCTIVE PULMONARY DISEASE.

• Grant number: 23/13626-7
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: November 01, 2023
• End date: October 31, 2024
• Field of knowledge: Biological Sciences - Physiology - Physiology of Organs and Systems
• Principal Investigator: Fernanda Degobbi Tenorio Quirino dos Santos Lopes
• Grantee: Ana Luiza Porfirio da Silva
• Host Institution: Faculdade de Medicina (FM). Universidade de São Paulo (USP). São Paulo , SP, Brazil
• Associated research grant: 18/02537-5 - Characterization of asthma-COPD overlap syndrome (ACOS): experimental and clinical studies, AP.TEM

Abstract

Asthmatic patients present a Th2 response profile, with an increase in cells TCD4+ and increased eosinophils around the airways, while COPD patients present a Th1 immune response profile, an increase in CD8+ T cells and an increase of neutrophils (Brusselle et al., 2011; Cosio et al., 2009). Our group has already demonstrated studies in an experimental model of COPD and in tissues from COPD patients, the role of TH17 response in the progression of this disease with a decrease in Tregulatory cells (Sales et al., 2017; Ito et al., 2019; Lorenço et al., 2021). In this sense, we will aim in this study, we evaluate the role of immunity in an overlapping model of asthma and COPD adaptive in this process, more specifically evaluating the role of the Th17 and Th2 response in this process, as well as the involvement of intracellular STAT proteins (from English "Activator of Transcription Protein") in the differentiation of naïve CD4+ T cells into their subtypes. Six male BALB/c mice distributed into four groups will be used.experimental: SF group: inhalations with sterile saline solution; OVA group: inhalations with ovalbumin solution (Toledo et al., 2013); ALS group: intratracheal instillations of elastase (Ikeda et al., 2014); ACOS group: inhalations with OVA solution and instillations intratracheal elastase injections (joining the previous protocols). The extracted lung tissue will be fixed and stained using an immunohistochemical technique for the following antibodies: Stat3, p-Stat3, Stat5, IL-17, FOXP3. Assessment of the expression of these markers in sections histological tests will be using the point counting morphometry technique (Weibel, 1963) with the reticle with 100 points and 50 straight lines attached to the microscope eyepiece (Toledo et al., 2013; Righetti et al., 2014; Theodoro-Júnior et al., 2017). Statistical analyzes will be performed using Sigma Stat 11.0 software. The Shapiro-Wilk test will be used to determine the distribution of the data. Depending on data distribution: an analysis of one-way variance followed by the Tukey or Kruskal-Wallis post hoc test followed by the Dunns test will be used to compare the groups. A value of p<0.05 will be considered statistically significant.Sales, D. S. et al. Regulatory T-Cell Distribution within Lung Compartments in COPD. COPDJ. Chronic Obstr. Pulm. Dis. 14, 533-542 (2017).Ito, J. T. et al. Th17/Treg imbalance in COPD progression: A temporal analysis using a CS-induced model. PLoS One (2019) doi:10.1371/journal.pone.0209351Ito JT, De Brito Cervilha DA, Lourenço JD, Gonçalves NG, Volpini RA, Caldini EG, et al.Th17/Treg imbalance in COPD progression: A temporal analysis using a CS-induced model.PLoS One. 2019;14(1):1-19.Lourenço JD, Ito JT, Martins M de A, Tibério I de FLC, Lopes FDTQ dos S. Th17/TregImbalance in Chronic Obstructive Pulmonary Disease: Clinical and Experimental Evidence.Front Immunol. 2021;12(December):1-11.Brusselle, G.G., et al. New insights into the immunology of chronic obstructive pulmonarydisease. Lancet, 2011. 378(9795): p. 1015-26.Cosio, M.G., et al. Immunologic aspects of chronic obstructive pulmonary disease. N Engl JMed, 2009. 360(23): p. 2445-54.Toledo, A.C., et al. Flavonone treatment reverses airway inflammation and remodelling inan asthma murine model. Br J Pharmacol, 2013. 168(7): p. 1736-49.Ikeda, G., et al. Effect of a cysteinyl leukotriene receptor antagonist on experimentalemphysema and asthma combined with emphysema. Am J Respir Cell Mol Biol, 2014.50(1): p. 18-29.Theodoro-Júnior OA, et al. A Plant Proteinase Inhibitor from Enterolobiumcontortisiliquum Attenuates Pulmonary Mechanics, Inflammation and Remodeling Inducedby Elastase in Mice. Int J Mol Sci. 2017 ,18(2).