Bivalent intranasal vaccine using influenza virus expressing SARS-CoV-2 protein S (spike): protection mechanisms and lung injury

• Grant number: 20/05527-0
• Support Opportunities: Regular Research Grants
• Duration: July 01, 2020 - June 30, 2022
• Field of knowledge: Biological Sciences - Immunology - Cellular Immunology
• Principal Investigator: Ricardo Tostes Gazzinelli
• Grantee: Ricardo Tostes Gazzinelli
• Host Institution: Faculdade de Medicina de Ribeirão Preto (FMRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil
• Associated researchers: Alexandre de Magalhães Vieira Machado ; João Santana da Silva ; Luciana Benevides ; Luiz Tadeu Moraes Figueiredo
• Associated research grant: 16/23618-8 - Immunological mechanisms of resistance and disease in malaria, AP.TEM
• Associated scholarship(s): 21/00930-4 - Evaluation of pulmonary tissue damage in viral infection (SARS-CoV-2 and/or Influenza), BP.PD

Abstract

Our proposal will consist of the evaluation of the mechanisms of the disease in models of pulmonary comorbidities challenged with SARS-CoV-2 and protection induced by a vaccine using a recombinant influenza virus expressing a segment of the Spike protein. We hypothesis is that the pathology triggered by excessive activation of the innate immune response in the airway mucosa and that with the development of the acquired response (IgA and cytotoxic T lymphocytes) induced by vaccination, the replication of the virus is minimized and the activation of cells of the innate immune system is attenuated. It is known that COVID-19 worsens in individuals with pulmonary comorbidities. However, it is not known whether this process involves a weakening of the immune response and an increase in viral load or just a worsening of the inflammatory reaction and, consequently, an increase in lung damage. As a model of comorbidities, we chose models widely used in our laboratories that trigger different pulmonary inflammatory response processes. Specifically, the models used will be infection models with fungus (Paracoccidioides brasiliensis, Pb), bacteria (Streptococcus pneumoniae, pneumococcus), viral (Influenza) and an asthma model, which induce chronic inflammation of the Th17 type and rich in neutrophils, a model of chronic pro-inflammatory inflammation rich in monocytes, acute pro-inflammatory response rich in monocytes and neutrophils and chronic Th2 response rich in eosinophils, respectively. In these models of comorbidities, different parameters will be evaluated, such as viral load, quality of the inflammatory infiltrate, activation of different pathways of the innate immune system, production of cytokines and lung injury. These studies will evaluate the toll-like receptors (TLR), inflammasomes and type I IFN pathways, using MyD88, ASC and type I IFN (IFNAR) deficient mice, respectively. Next, we will assess the ability of the PR8 influenza virus that is defective for multiplication because it does not express neuraminidase. In this virus, the extracellular region of neuroamindase is replaced by the RBD domain of the SARS-CoV-2 Spike protein. Studies carried out by our group have already demonstrated the ability of this neuraminidase-deficient virus to provide 100% protection for challenged mice. Therefore, we believe that we have a bivalent vaccine against influenza and SARS-CoV-2. Besides, in previous studies, we built an influenza virus that expresses the PspA protein of pneumococcus and protects against co-infection with influenza and S. Pneumoniae, the major cause of lethality in patients with influenza and superinfected patients. Therefore, we will evaluate the degree of protection and immunological mechanisms involved in the viral load and inflammation of mice vaccinated and challenged with SARS-CoV-2, and in some groups, infected concomitantly with competent influenza-replication or pneumococcus. As for the protective mechanism induced by the vaccine, we will study the level of protection in mice deficient in B lymphocytes, CD8 T lymphocytes, and combined immunodeficiency, using the mutant mice for ¼ chain, b2-microglobulin mice and RAG knockouts (KO) mice. The focus will be given to the role of neutralizing antibodies and IgA produced in the mucosa, as well as cellular response mediated by CD4 Th1 and CD8 cytotoxic lymphocytes. We believe that these studies will lead to a better understanding of the mechanisms of the disease and protection against influenza and SARS-CoV-2 viruses. (AU)

Scientific publications

(5)

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

FUMAGALLI, MARCILIO JORGE; CASTRO-JORGE, LUIZA ANTUNES; DE SOUZA, WILLIAM MARCIEL; DE AZEVEDO, PATRICK ORESTES; HANSEN, ALANA WITT; GAZZINELLI, RICARDO TOSTES; DA FONSECA, BENEDITO ANTONIO LOPES; SPILKI, FERNANDO ROSADO; FIGUEIREDO, LUIZ TADEU MORAES. oronaVac and ChAdOx1 Vaccination and Gamma Infection Elicited Neutralizing Antibodies against the SARS-CoV-2 Delta Varian. IRUSES-BASE, v. 14, n. 2, FEB 2022. (18/09383-3, 20/05527-0, 19/26119-0)

FUMAGALLI, MARCILIO JORGE; CASTRO-JORGE, LUIZA ANTUNES; FRAGA-SILVA, THAIS FERNANDA DE CAMPOS; DE AZEVEDO, PATRICK ORESTES; CAPATO, CARLOS FABIANO; RATTIS, BRUNA AMANDA CRUZ; HOJO-SOUZA, NATALIA SATCHIKO; FLORIANO, VITOR GONCALVES; DE CASTRO, JULIA TEIXEIRA; RAMOS, SIMONE GUSMAO; et al. Protective Immunity against Gamma and Zeta Variants after Inactivated SARS-CoV-2 Virus Immunization. Viruses-Basel, v. 13, n. 12, DEC 2021. (19/26119-0, 20/05527-0, 18/09383-3, 19/09881-6, 19/11213-1)

ASSIS, BRUNA RODRIGUES DIAS; GOMES, ISABELA PEREIRA; DE CASTRO, JULIA TEIXEIRA; RIVELLI, GRAZIELLA GOMES; DE CASTRO, NATALIA SALAZAR; GOMEZ-MENDOZA, DIANA PAOLA; BAGNO, FLAVIA FONSECA; HOJO-SOUZA, NATALIA SATCHIKO; MAIA, ANA LUIZA CHAVES; LAGES, EDUARDO BURGARELLI; et al. Quality attributes of CTVad1, a nanoemulsified adjuvant for phase I clinical trial of SpiN COVID-19 vaccine. Nanomedicine, v. 18, n. 18, p. 20-pg., 2023-09-15. (20/05527-0)

CASTRO, JULIA T.; AZEVEDO, PATRICK; FUMAGALLI, MARCILIO J.; HOJO-SOUZA, NATALIA S.; SALAZAR, NATALIA; ALMEIDA, GREGORIO G.; OLIVEIRA, LIVIA I.; FAUSTINO, LIDIA; ANTONELLI, LIS R.; MARCAL, TOMAS G.; et al. Promotion of neutralizing antibody-independent immunity to wild-type and SARS-CoV-2 variants of concern using an RBD-Nucleocapsid fusion protein. NATURE COMMUNICATIONS, v. 13, n. 1, p. 16-pg., 2022-08-17. (20/05527-0)

AZEVEDO, PATRICK O.; HOJO-SOUZA, NATALIA S.; FAUSTINO, LIDIA P.; FUMAGALLI, MARCILIO J.; HIRAKO, ISABELLA C.; OLIVEIRA, EMILIANO R.; FIGUEIREDO, MARIA M.; CARVALHO, ALEX F.; DORO, DANIEL; BENEVIDES, LUCIANA; et al. Differential requirement of neutralizing antibodies and T cells on protective immunity to SARS-CoV-2 variants of concern. NPJ VACCINES, v. 8, n. 1, p. 17-pg., 2023-02-13. (20/05527-0)