Rhinovirus in human lymphoid tissues: investigation of virus-cell interaction and viral persistence

• Grant number: 17/25654-4
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• Effective date (Start): March 01, 2018
• Effective date (End): February 28, 2021
• Field of knowledge: Biological Sciences - Microbiology
• Principal Investigator: Eurico de Arruda Neto
• Grantee: Ronaldo Bragança Martins Júnior
• Host Institution: Faculdade de Medicina de Ribeirão Preto (FMRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil

Abstract

Rhinovirus (RV) is the main causative agent of the so called "common cold", the most frequent viral illness of mankind. RV infections also are frequently associated with asthma, exacerbations of chronic bronchitis and emphysema, as well as with secondary complications such as acute sinusitis and otitis media. Additionally, RV is frequently detected in tonsillar tissues, and in nasopharyngeal secretions from patients with chronic adenotonsillar diseases, even in the absence of symptoms of acute respiratory infections. Evidence generated by our research group indicate that the main phenotypes of cells infected by RV in lymphoid tissues are reticular epithelial cells, lymphoid CD4+ T cells, and B lymphocytes.This project, firmly based on preliminary data from our research group, aims at investigating the systemic nature of RV infections in human lymphoid tissues, and the virus-host interaction comparing lymphomononuclear and fibro-epithelial cells from human tonsils infected with RV in vitro. We will study the activation of regulatory genes involved in inflammation and antiviral response in primary cultures of lymphomononuclear and fibro-epithelial cells infected with RV. We will also evaluate the expression. Of miRNAs of proinflammatory and anti-inflammatory cytokines in RV-infected cells, as well as the impact of RV infection in cell viability and immunoglobulin synthesis in susceptible B cells. Lastly, we will study the inflammasomes and production of markers of persistent viral infections (IL-10 e PD-1) in primary cultures of lymphomononuclear and fibro-epithelial cells from human tonsils infected with RV in vitro.Evidence produced by our group indicates that the main RV-infected phenotypes in these tissues are reticular epithelial cells, CD4 + T lymphocytes and B cells. This project, based on preliminary data, proposes to investigate the infection of rhinovirus in lymphomonuclear cells (LMNC) and fibroepithelial cells derived from secondary human lymphoid tissues to comparatively elucidate the host virus-cell interaction against RV infection.To this end, the magnitude of activation of transcription factors of regulatory genes involved in inflammation and antiviral response in infected primary cultures of fibroepithelial cells and LMNC will be determined. We intend to evaluate the balance of mRNA expression of pro-inflammatory and anti-inflammatory cytokines in these cells infected with VR strains, as well as to verify the impact of VR infection on cell viability and synthesis of immunoglobulins in the B-cell susceptible subpopulation specifically. Finally, we want to study the kinetics of the assembly of inflammatories between the different cell lines obtained from lymphoid tissues and to analyze markers of persistent viral infection (IL-10 and PD-1) in primary cultures infected in vitro.