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Two-Step In Vitro Model to Evaluate the Cellular Immune Response to SARS-CoV-2

2021; Multidisciplinary Digital Publishing Institute; Volume: 10; Issue: 9 Linguagem: Inglês

10.3390/cells10092206

2073-4409

Juliana Gil Melgaço, Tamiris Azamor, Andréa Marques Vieira da Silva, José Linhares, Tiago Pereira Santos, Ygara S. Mendes, Sheila Maria Barbosa de Lima, Camilla Bayma Fernandes, Jane da Silva, Alessandro Fonseca de Souza, Luciana Tubarão, Danielle Brito e Cunha, Tamires Bomfim Santos Pereira, Catarina Eugênia Menezes, Milene Dias Miranda, Aline da Rocha Matos, Bráulia Costa Caetano, Jéssica Santa Cruz Carvalho Martins, Thyago Leal-Calvo, Natália Fintelman-Rodrigues, Carolina Q. Sacramento, Marilda Mendonça Siqueira, Milton Ozório Moraes, Sotiris Missailidis, Patrícia Neves, Ana Paula Dinis Ano Bom,

Long-Term Effects of COVID-19

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.