Two-Step In Vitro Model to Evaluate the Cellular Immune Response to SARS-CoV-2
2021
Juliana G. Melgaço | Tamiris Azamor | Andréa M. V. Silva | José Henrique R. Linhares | Tiago P. dos Santos | Ygara S. Mendes | Sheila M. B. de Lima | Camilla Bayma Fernandes | Jane da Silva | Alessandro F. de Souza | Luciana N. Tubarão | Danielle Brito e Cunha | Tamires B. S. Pereira | Catarina E. L. Menezes | Milene D. Miranda | Aline R. Matos | Braulia C. Caetano | Jéssica S. C. C. Martins | Thyago L. Calvo | Natalia F. Rodrigues | Carolina Q. Sacramento | Marilda M. Siqueira | Milton O. Moraes | Sotiris Missailidis | Patrícia C. C. Neves | Ana Paula D. Ano Bom
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<sup>+</sup> 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.
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