Abstract
Since neutrophil extracellular traps formation (NET-osis) can be assessed indirectly by treating healthy neutrophils with blood-derived fluids from patients and then measuring the NETs response, we designed a pilot study to convey high-dimensional cytometry of peripheral blood immune cells and cytokines, combined with clinical features, to understand if NET-osis assessment could be included in the immune risk profiling to early prediction of clinical patterns, disease severity, and viral clearance at 28 days in COVID-19 patients. Immune cells composition of peripheral blood, cytokines concentration and in-vitro NETosis were detected in peripheral blood of 41 consecutive COVID-19 inpatients, including 21 mild breakthrough infections compared to 20 healthy donors, matched for sex and age. Major immune dysregulation in peripheral blood in not-vaccinated COVID-19 patients compared to healthy subjects included: a significant reduction of percentage of unswitched memory B-cells and transitional B-cells; loss of naïve CD3 + CD4 + CD45RA + and CD3 + CD8 + CD45RA + cells, increase of IL-1β, IL-17A and IFN-γ. Myeloid compartment was affected as well, due to the increase of classical (CD14 ++ CD16 – ) and intermediate (CD14 ++ CD16 + ) monocytes, overexpressing the activation marker CD64, negatively associated to the absolute counts of CD8+ CD45R0+ cells, IFN-γ and IL-6, and expansion of monocytic-like myeloid derived suppressor cells. In not-vaccinated patients who achieved viral clearance by 28 days we found at hospital admission lower absolute counts of effector cells, namely CD8 + T cells, CD4 + T-cells and CD4 + CD45RO + T cells. Percentage of in-vitro NET-osis induced by patients’ sera and NET-osis density were progressively higher in moderate and severe COVID-19 patients than in mild disease and controls. The percentage of in-vitro induced NET-osis was positively associated to circulating cytokines IL-1β, IFN-γ and IL-6. In breakthrough COVID-19 infections, characterized by mild clinical course, we observed increased percentage of in-vitro NET-osis, higher CD4+ CD45RO+ and CD8+ CD45RO+ T cells healthy or mild-COVID-19 not-vaccinated patients, reduced by 24 h of treatment with ACE inhibitor ramipril. Taken together our data highlight the role of NETs in orchestrating the complex immune response to SARS-COV-2, that should be considered in a multi-target approach for COVID-19 treatment.
【초록키워드】 COVID-19, Treatment, Monocytes, immune response, Cytokines, IL-17A, T cells, IL-6, disease severity, neutrophil, T-cells, Sex, risk, cytokine, CD4, CD8, viral clearance, immune, Clinical features, Peripheral blood, COVID-19 treatment, T cell, Clinical course, cells, sera, Patient, Neutrophil extracellular trap, NETosis, Mild, breakthrough infections, immune cells, age, Hospital admission, Breakthrough infection, expansion, T-cell, inhibitor, disease, moderate, patients, CD45RO, NETs, myeloid, COVID-19 patients, IFN-γ, Mild disease, marker, CD16, IL-1β, Concentration, ACE, Immune cell, COVID-19 patient, Cytometry, In-vitro, immune dysregulation, memory B-cell, dysregulation, ramipril, Major, Inpatients, reduction, Activation, healthy donors, pilot study, healthy subjects, COVID-19 infections, complex, naïve, CD4+, CD14, CD45, CD45RA, CD64, CD8+, effector cells, circulating cytokines, NET, CD3, approach, controls, Cell, highlight, classical, affected, healthy, reduced, characterized, healthy subject, circulating cytokine, overexpressing, percentage, severe COVID-19 patient, 【제목키워드】 Clinical course, Patient, Breakthrough infection, COVID-19 sera,