1. Two-Step In Vitro Model to Evaluate the Cellular Immune Response to SARS-CoV-2.
- Author
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Melgaço JG, Azamor T, Silva AMV, Linhares JHR, Dos Santos TP, Mendes YS, de Lima SMB, Fernandes CB, da Silva J, de Souza AF, Tubarão LN, Brito E Cunha D, Pereira TBS, Menezes CEL, Miranda MD, Matos AR, Caetano BC, Martins JSCC, Calvo TL, Rodrigues NF, Sacramento CQ, Siqueira MM, Moraes MO, Missailidis S, Neves PCC, and Ano Bom APD
- Subjects
- Adult, Alveolar Epithelial Cells virology, COVID-19 blood, COVID-19 genetics, Cytokines genetics, Cytokines metabolism, Female, Gene Expression Regulation, Humans, Immunologic Memory immunology, Killer Cells, Natural immunology, Leukocytes, Mononuclear virology, Male, Middle Aged, Phenotype, T-Lymphocytes immunology, Virus Replication physiology, Young Adult, COVID-19 immunology, COVID-19 virology, Immunity, Cellular, Models, Biological, SARS-CoV-2 physiology
- Abstract
The cellular immune response plays an important role in COVID-19, caused by SARS-CoV-2. This feature makes use of in vitro models' useful tools to evaluate vaccines and biopharmaceutical effects. Here, we developed a two-step model to evaluate the cellular immune response after SARS-CoV-2 infection-induced or spike protein stimulation in peripheral blood mononuclear cells (PBMC) from both unexposed and COVID-19 (primo-infected) individuals (Step1). Moreover, the supernatants of these cultures were used to evaluate its effects on lung cell lines (A549) (Step2). When PBMC from the unexposed were infected by SARS-CoV-2, cytotoxic natural killer and nonclassical monocytes expressing inflammatory cytokines genes were raised. The supernatant of these cells can induce apoptosis of A549 cells (mock vs. Step2 [mean]: 6.4% × 17.7%). Meanwhile, PBMCs from primo-infected presented their memory CD4
+ T cells activated with a high production of IFNG and antiviral genes. Supernatant from past COVID-19 subjects contributed to reduce apoptosis (mock vs. Step2 [ratio]: 7.2 × 1.4) and to elevate the antiviral activity (iNOS) of A549 cells (mock vs. Step2 [mean]: 31.5% × 55.7%). Our findings showed features of immune primary cells and lung cell lines response after SARS-CoV-2 or spike protein stimulation that can be used as an in vitro model to study the immunity effects after SARS-CoV-2 antigen exposure.- Published
- 2021
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