Dados do Trabalho

Título

Analysis of the Immunomodulatory Effects of Cytokine Blockers on Healthy PBMCs Treated with Serum from COVID-19 Patients

Introdução

SARS-CoV-2 infection causes COVID-19 with varied clinical presentations and immunopathologies. The cytokine storm interferes with viral evasion and cellular immune responses. The use of immunomodulators aims to reduce the inflammation caused by the disease.

Objetivo (s)

To evaluate the immunomodulatory effect of cytokine blockers on healthy PBMCs treated with pooled serum from COVID-19 patients.

Material e Métodos

PBMCs from healthy individuals were cultured (2x10^6 cells/mL) and treated with: (1) pool serum from six severe COVID-19 patients; (2) pool serum from six healthy donors from 2018 (pre-pandemic); (3) pool serum from four post-COVID-19 patients; (4) 0.9% saline solution. The cultures received cytokine-blocking antibodies: Infliximab and Adalimumab 25 µg/mL (anti-TNF-α); Tocilizumab 100 µg/mL (anti-IL-6); and Secukinumab 25 µg/mL (anti-IL-17). After incubation for 72 hours at 37°C and 5% CO2, the supernatant was collected, and the cytokines TNF-α, IL-6, IL-10, IFN-γ, and IL-17 were measured by ELISA (BD BIOSCIENCES, USA) according to the manufacturer's instructions. Approved by the National Research Ethics Committee: CAEE 30474020.2.0000.0008.

Resultados e Conclusão

TNF-α levels are elevated in COVID-19, with its release occurring during the binding of SARS-CoV-2 to the angiotensin-converting enzyme (ACE), facilitating viral entry and tissue damage due to elevated levels (1,2). In our results, treatment with Infliximab in cells treated with COVID-19 serum pool showed a reduction in IL-6, IL-10, IL-17, and TNF-α levels. Treatment with Adalimumab, which is also anti-TNF-α, showed a reduction in TNF-α and IL-10 levels. Elevated IL-6 levels are associated with poor prognosis in COVID-19 patients (3,4). Our results showed that treatment with Tocilizumab in cells treated with COVID-19 serum pool demonstrated a reduction in IFN-γ, IL-10, IL-1β, and IL-6 levels. In SARS-CoV-2 infection, T-helper-17 (Th17) cells lead to excessive IL-17 release, with its role being protective or driving hyperinflammation (5–7). Treatment with Secukinumab and COVID-19 serum pool reduced IL-17 and IL-1β levels. Interfering with cytokine signaling using immunomodulators can significantly reduce COVID-19 hyperinflammation. Considering the data, this study revealed possible modulations that could help reduce serum cytokine levels, contributing to a better prognosis. Such evidence may aid in addressing future challenges.

Palavras Chave

COVID-19; : Cytokines; Serum; Immunomodulatory; Cytokine Blockers; PBMC's.