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HIGH LEVELS OF PRO-INFLAMMATORY SARS-COV-2- SPECIFIC BIOMARKERS REVEALED BY IN VITRO WHOLE BLOOD CYTOKINE RELEASE ASSAY (CRA) IN RECOVERED AND LONG-COVID-19 PATIENTS
Author
Gomes, Shayane Martins Rodrigues
Brito, Andréia Carolinne de Souza
Manfro, Wânia Ferraz Pereira
Ribeiro-Alves, Marcelo
Ribeiro, Roberto Stefan de Almeida
Cal, Mariana Soares da
Lisboa, Vinicius da Cunha
Abreu, Daniel Paiva Barros de
Castilho, Leda dos Reis
Porto, Luís Cristóvão de Moares Sobrino
Mafort, Thiago Thomáz
Lopes, Agnaldo José
Silva, Silvia Amaral Gonçalves da
Dutra, Patrícia Maria Lourenço
Rodrigues, Luciana Silva
Brito, Andréia Carolinne de Souza
Manfro, Wânia Ferraz Pereira
Ribeiro-Alves, Marcelo
Ribeiro, Roberto Stefan de Almeida
Cal, Mariana Soares da
Lisboa, Vinicius da Cunha
Abreu, Daniel Paiva Barros de
Castilho, Leda dos Reis
Porto, Luís Cristóvão de Moares Sobrino
Mafort, Thiago Thomáz
Lopes, Agnaldo José
Silva, Silvia Amaral Gonçalves da
Dutra, Patrícia Maria Lourenço
Rodrigues, Luciana Silva
Affilliation
Rio de Janeiro State University. Parasitology and Immunology. Medical Science Faculty. Department of Microbiology. Rio de Janeiro, RJ, Brazil.
Rio de Janeiro State University. Parasitology and Immunology. Medical Science Faculty. Department of Microbiology. Rio de Janeiro, RJ, Brazil.
Rio de Janeiro State University. Parasitology and Immunology. Medical Science Faculty. Department of Microbiology. Rio de Janeiro, RJ, Brazil.
Oswaldo Cruz Foundation. National Institute of Infectology Evandro Chagas. Laboratory of Clinical Research on STD/AID. Rio de Janeiro, RJ, Brazil.
Rio de Janeiro State University. Department of General Pathology and Laboratories. Laboratory of Immunopathology. Rio de Janeiro, RJ, Brazil.
Pedro Ernesto University Hospital. Policlínica Piquet Carneiro. Pulmonary and Tisiology Department. Rio de Janeiro, RJ, Brazil.
Rio de Janeiro State University. Department of General Pathology and Laboratories. Laboratory of Immunopathology. Rio de Janeiro, RJ, Brazil.
Federal University of Rio de Janeiro. Chemical Engineering Program, Cell Culture Engineering Lab (COPPE). Rio de Janeiro, RJ, Brazil.
Federal University of Rio de Janeiro. Chemical Engineering Program, Cell Culture Engineering Lab (COPPE). Rio de Janeiro, RJ, Brazil.
Rio de Janeiro State University. Department of Histology and Embryology Institute of Biology Roberto Alcantara Gomes. Histocompatibility and Cryopreservation Laboratory. Rio de Janeiro, RJ, Brazil.
Pedro Ernesto University Hospital. Policlínica Piquet Carneiro. Pulmonary and Tisiology Department. Rio de Janeiro, RJ, Brazil.
Pedro Ernesto University Hospital. Policlínica Piquet Carneiro. Pulmonary and Tisiology Department. Rio de Janeiro, RJ, Brazil.
Rio de Janeiro State University. Parasitology and Immunology. Medical Science Faculty. Department of Microbiology. Rio de Janeiro, RJ, Brazil.
Rio de Janeiro State University. Parasitology and Immunology. Medical Science Faculty. Department of Microbiology. Rio de Janeiro, RJ, Brazil.
Rio de Janeiro State University. Department of General Pathology and Laboratories. Laboratory of Immunopathology. Rio de Janeiro, RJ, Brazil.
Rio de Janeiro State University. Parasitology and Immunology. Medical Science Faculty. Department of Microbiology. Rio de Janeiro, RJ, Brazil.
Rio de Janeiro State University. Parasitology and Immunology. Medical Science Faculty. Department of Microbiology. Rio de Janeiro, RJ, Brazil.
Oswaldo Cruz Foundation. National Institute of Infectology Evandro Chagas. Laboratory of Clinical Research on STD/AID. Rio de Janeiro, RJ, Brazil.
Rio de Janeiro State University. Department of General Pathology and Laboratories. Laboratory of Immunopathology. Rio de Janeiro, RJ, Brazil.
Pedro Ernesto University Hospital. Policlínica Piquet Carneiro. Pulmonary and Tisiology Department. Rio de Janeiro, RJ, Brazil.
Rio de Janeiro State University. Department of General Pathology and Laboratories. Laboratory of Immunopathology. Rio de Janeiro, RJ, Brazil.
Federal University of Rio de Janeiro. Chemical Engineering Program, Cell Culture Engineering Lab (COPPE). Rio de Janeiro, RJ, Brazil.
Federal University of Rio de Janeiro. Chemical Engineering Program, Cell Culture Engineering Lab (COPPE). Rio de Janeiro, RJ, Brazil.
Rio de Janeiro State University. Department of Histology and Embryology Institute of Biology Roberto Alcantara Gomes. Histocompatibility and Cryopreservation Laboratory. Rio de Janeiro, RJ, Brazil.
Pedro Ernesto University Hospital. Policlínica Piquet Carneiro. Pulmonary and Tisiology Department. Rio de Janeiro, RJ, Brazil.
Pedro Ernesto University Hospital. Policlínica Piquet Carneiro. Pulmonary and Tisiology Department. Rio de Janeiro, RJ, Brazil.
Rio de Janeiro State University. Parasitology and Immunology. Medical Science Faculty. Department of Microbiology. Rio de Janeiro, RJ, Brazil.
Rio de Janeiro State University. Parasitology and Immunology. Medical Science Faculty. Department of Microbiology. Rio de Janeiro, RJ, Brazil.
Rio de Janeiro State University. Department of General Pathology and Laboratories. Laboratory of Immunopathology. Rio de Janeiro, RJ, Brazil.
Abstract
Background: Cytokines induced by SARS-CoV-2 infection play a crucial role in the pathophysiology of COVID-19 and hyperinflammatory responses have been associated with poor clinical outcomes, with progression to severe conditions or long-term subacute complications named as long-COVID-19. Methods: In this cross-sectional study, we aimed to evaluate a set of antigen-specific inflammatory cytokines in blood from recovered COVID-19 individuals or who suffered a post-acute phase of SARS-CoV-2 infection compared to healthy individuals with no history of COVID-19 exposition or infection. Interferon-gamma (IFN-γ), IFN-γ-induced protein 10 (IP-10), tumor necrosis factor (TNF), IL-1β, IL-2, IL-4, IL-6, IL-8, IL-10, IL-12, and IL-17A were quantified by multiplex cytometric bead assay and enzyme-linked immunosorbent assay after stimulation of whole blood with recombinant Spike protein from SARS-CoV-2. Additionally, all participants have evaluated for anti-(S) protein-specific IgG antibodies. Clinical specimens were collected within two months of COVID-19 diagnosis. Results: A total of 47 individuals were enrolled in the study, a median age of 43 years (IQR = 14.5), grouped into healthy individuals with no history of infection or exposure to SARS-CoV-2 (unexposed group; N = 21); and patients from the Health Complex of the Rio de Janeiro State University (UERJ), Brazil, who were SARS-CoV-2 positive by RT-PCR (COVID-19 group)-categorized as recovered COVID-19 (N = 11) or long-COVID-19 (N = 15). All COVID-19 patients presented at least one signal or symptom during the first two weeks of infection. Six patients were hospitalized and required invasive mechanical ventilation. Our results showed that COVID-19 patients had significantly higher levels of IFN-γ, TNF, IL-1β, IL-2, IL-6, IL-8, and IP-10 than the unexposed group. The long-COVID-19 group has presented significantly high levels of IL-1β and IL-6 compared to unexposed individuals, but not from recovered COVID-19. A principal-component analysis demonstrated 84.3% of the total variance of inflammatory-SARS-CoV-2 response in the first two components, and it was possible to stratify IL-6, TNF, IL-1β, IL-10, and IL-2 as the top-five cytokines which are candidates to discriminate COVID-19 group (including long-COVID-19 subgroup) and healthy unexposed individuals. Conclusion: We revealed important S protein-specific differential biomarkers in individuals affected by COVID-19, bringing new insights into the inflammatory status or SARS-CoV-2 exposition determination.
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