A Nutritional Blend Suppresses the Inflammatory Response from Bronchial Epithelial Cells Induced by SARS-CoV-2

Abstract

Even after virus elimination, numerous sequelae of coronavirus disease 2019 (COVID-19) persist. Based on accumulating evidence, large amounts of proinflammatory cytokines are released to drive COVID-19 progression, severity, and mortality, and their levels remain elevated after the acute phase of COVID-19, playing a central role in the disease’ sequelae. In this manner, bronchial epithelial cells are the first cells hyperactivated by severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2), leading to massive cytokine release, triggering the hyperactivation of leukocytes and other cells, and mediating COVID-19 sequelae. Therefore, proinflammatory cytokine production is initiated by the host. This in vitro study tested the hypothesis that ImmuneRecov™, a nutritional blend, inhibits the SARS-CoV-2-induced hyperactivation of human bronchial epithelial cells (BEAS-2B). BEAS-2B (5x10⁴/mL/well) cells were cocultivated with 1 ml of blood from a SARS-CoV-2-infected patient for 4 h, and the nutritional blend (1 µg/mL) was added in the first minute of coculture. After 4 h, the cells were recovered and used for analyses of cytotoxicity with the (3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide) (MTT) assay and the expression of the IL-1β, IL-6, and IL-10 mRNAs. The supernatant was collected to measure cytokine levels. SARS-CoV-2 incubation resulted in increased levels of IL-1β and IL-6 in BEAS-2B cells (p < 0.001). Treatment with the nutritional blend resulted in reduced levels of the proinflammatory cytokines IL-1β and IL-6 (p < 0.001) and increased levels of the anti-inflammatory cytokine IL-10 (p < 0.001). Additionally, the nutritional blend reduced the expression of the IL-1β and IL-6 mRNAs in SARS-CoV-2-stimulated cells and increased the expression of the IL-10 and IFN-γ mRNAs. In conclusion, the nutritional blend exerts important anti-inflammatory effects on cells in the context of SARS-CoV-2 infection.

Keywords:

• COVID-19
• cytokines
• epithelium
• inflammation
• SARS-CoV-2

Disclosure statement

CRO, JRM and RPV are the owners of the company GAP Biotech, the producer of ImmuneRecov™.

Data availability statement

All raw data are restricted but will be made available upon reasonable request.

Additional information

Funding

This study was partially supported by Sao Paulo Research Foundation (FAPESP) grant #2012/15165-2 and by GAP Biotech.

Notes on contributors

José Roberto Mateus-Silva

José Roberto Mateus-Silva holds a degree in Systems Analysis and Development from UNIP, a postgraduate degree in Biotechnology from Unyleya, an MBA in Management, Innovation and Health Services from PUC-RS (in progress), a degree in Medicine from UAM (in progress). He is the CEO of GAP Biotech, a company specialized in bioinformatics founded in 1983. He is the founder of Grupo GAP Networks, an IT company dedicated to the production of specialist software, where he has established offices in Argentina, Brazil, Chile, Mexico and Panama, to serve multinational companies around the world. He is the founder of DAPP, a company specializing in application development. He is the founder of ACEMT-philanthropic entity that maintains Escola Madre Teresa, an elementary school that serves children in rural areas of São José dos Campos, especially those in a situation of social vulnerability. In addition to the new degrees in progress, it has invested its time in projects that are in line with the 17 Sustainable Development Goals (SDGs) established at the UN Conference on sustainable development in Rio de Janeiro in 2012, notably the foundation of the Sophia Institute of Science, Technology and Innovation, scheduled to start operations in February 2021.

Carlos Rocha Oliveira

Carlos Rocha Oliveira, Pharmacist, Master in Pharmacology and Doctor in Biotechnology (USP). Postdoctoral fellow at the Institute of Pharmacology and Molecular Biology (UNIFESP). Professor of the Medicine course at Universidade Anhembi Morumbi and Coordinator of the Ethics Committee in Human Research at Universidade Anhembi Morumbi (CEP-UAM). He develops projects from compounds of natural origin, with an emphasis on cell signaling and the mode of action of drugs. Advisor at the Graduate Program in Biomedical Engineering (UNIFESP), Campus São José dos Campos-SP.

Maysa Alves Rodrigues Brandao-Rangel

Maysa Alves Rodrigues Brandao-Rangel, Graduated in Physiotherapy from Faculdade do Clube Náutico Mogiano (1997–2000), Improvement in Motor and Respiratory Physiotherapy at Hospital do Servidor Público Estadual (2000), Hospital Improvement at Santa Casa de São Paulo (2001), Postgraduate in Pediatric Physiotherapy (2003–2004) at Faculdades Metropolitanas Unidas de São Paulo, Graduated in Physiotherapy in Orthopedics and Traumatology at Universidade Gama Filho (2006–2007). Physiotherapist at Hospital Policlin (2001-2003), Physiotherapist responsible for the Fisiovale clinic (2001–2009) working in the area of orthopedics and traumatology, Physiotherapist at the Municipal Hospital Dr José de Carvalho Florence (2009-until August 2016), working in the Adult ICU, Adult and Emergency Infirmary, contributor to the protocols for ventilatory weaning, non-invasive mechanical ventilation and tracheostomy weaning. Master in Medicine at Universidade Nove de Julho, PhD student in Rehabilitation and Human Movement Sciences at the Federal University of São Paulo, student at the Laboratory of Pulmonary Immunology and Exercise (LABPEI), in the city of São José dos Campos. from the Brazilian Institute of Teaching and Research in Pulmonary and Exercise Immunology (IBEPIPE) and student at the UNIFESP laboratory who develops clinical and experimental research work, focusing on cellular and molecular immunological aspects involved in both the pathophysiology of lung diseases and exercise and its interaction in the prevention and treatment of pulmonary and inflammatory diseases. Currently also active in the Post COVID-19 Rehabilitation research project at UNIFESP-SJC.

Anamei Silva-Reis

Anamei Silva-Reis, Graduated in Physical Education from the State University of Pará (UEPA 2015–2018). During her graduation period, she was a member of the Exercise Biochemistry Laboratory (LABEX) and the Resistance Exercise and Health Laboratory (LERES), both with an emphasis on Parkinson’s Disease. She has a PIBIC / FAPESPA Scientific Initiation bond during the 2017/2018 period. She is currently a member of the Laboratory of Pulmonary Immunology and Exercise (LABPEI), with an emphasis on the pathophysiology of lung diseases and the effects of physical exercise.

Fabiana Regina da Silva Olimpio

Fabiana Regina da Silva Olimpio, PhD student and Master of Science from the Postgraduate Program in Translational Medicine at the Federal University of São Paulo and a degree in Biological Sciences from the Universidade Paulista.

Lucas dos Santos Zamarioli

Lucas dos Santos Zamarioli, Graduated in Biological Sciences from Universidade Anhembi Morumbi (2018). Master by the Postgraduate Program in Pharmacology at Escola Paulista de Medicina, Federal University of São Paulo (EPM/UNIFESP), in the Department of Drug Action Mode (2021), and PhD candidate at the same program. Main activity in the areas of phytotherapy, cancer and muscle, with the objective of detecting possible mechanisms of action of plant extracts and isolated components in tumor cell lines (cytotoxicity) and muscle (cytoprotection).

Flavio Aimbire

Flavio Aimbire holds a degree in Biological Sciences from the University of the State of Rio de Janeiro (1996), a Master’s degree in Pharmacology from the University of São Paulo (2001) and a PhD in Biomedical Engineering from the University of Vale do Paraíba with a period at the University of Bergen, Norway (2006). He is currently a professor at the Federal University of São Paulo (UNIFESP) and researcher/collaborator in the postgraduate program in Biotechnology, and researcher/advisor in the postgraduate program in Translational Medicine at Unifesp. He has experience in the area of Pulmonary Inflammation and Immunology, with emphasis on bronchial reactivity, leukocyte-endothelium interaction, cell signaling and mechanism of action, working mainly on the following topics: asthma, chronic obstructive pulmonary disease, intestinal ischemia and reperfusion, sepsis, and therapies anti-inflammatory.

Rodolfo P. Vieira

Rodolfo P. Vieira, Graduated in Physical Education (Univap 1998–2001), Master in Biological Sciences (Univap 2001–2003), Doctorate in Pathology (CAPES Scholarship, USP School of Medicine 2004–2007) and Post-Doctorate in Physical Activity and Pulmonary Immunopathology (FAPESP Scholarship, Faculdade de Medicina USP 2008-2009), Post-Doctorate in Asthma Immunology (Deutsche Forchungsgemeinschaft Contract 2009-2010) from Albert-Ludwigs University of Freiburg - Germany, Post-Doctorate in Physical Activity and Pulmonary Immunopathology (European Respiratory Award Scholarship Society/Marie Curie Foundation/European Union 2010–2011). He was a visiting researcher at the Department of Pulmonology at the Albert-Ludwigs University of Freiburg in Germany (June 2010 until May 2011) and a research associate in the same department (June 2011 until December 2011). He is the researcher responsible for the project “Effects of aerobic physical training on lung changes induced by different products of air pollution” funded by the MCT / CNPq that is part of the (INCTs-National Institutes of Science and Technology). Since April/2014 he is the author and responsible for the Young Researcher Project FAPESP 2012/15165-2 entitled “Role of purinergic signaling and SOCS-JAK-STAT signaling in the anti-inflammatory effects of aerobic training in experimental models of asthma and in asthmatic individuals”, from where he obtained resources to set up the Laboratory of Pulmonary Immunology and Exercise (LABPEI) in the city of São José dos Campos. He is currently director of the Brazilian Institute of Teaching and Research in Pulmonary and Exercise Immunology (IBEPIPE). He is coordinator of LABPEI (www.labpei.com.br), which is the functional arm of IBEPIPE, developing clinical and experimental research works, focusing on cellular and molecular immunological aspects involved both in the pathophysiology of lung diseases and in the effects of physical exercise in lung diseases. Furthermore, research in the area of exercise immunology, which is carried out with professional and amateur athletes and different sports teams, is included in the laboratory’s research focus. Advisor of the master’s and doctoral programs in Human Movement and Rehabilitation Sciences at UNIFESP (and collaborating professor) and the professional master’s degree in Bioengineering at Universidade Brasil and the master’s and doctorate in Human Movement and Rehabilitation at Unievangélica and also the professional master’s degree in Pharmaceutical Sciences from Unievangelical. He has been a CNPq level 2 research productivity fellow since 2016.