Inactivation of SARS Coronavirus 2 and COVID-19 patient samples for contemporary immunology and metabolomics studies

Eddins, Devon J.; Bassit, Leda; Chandler, Joshua D.; Haddad, Natalie S.; Musall, Kathryn; Yang, Junkai; Kosters, Astrid; Dobosh, Brian; Hernandez, Mindy; Ramonell, Richard P.; Tirouvanziam, Rabindra; Lee, Frances Eun-Hyung; Zandi, Keivan; Schinazi, Raymond F.; Ghosn, Eliver

doi:10.1101/2021.10.22.465481

Cited by 2 publications

(3 citation statements)

References 68 publications

(86 reference statements)

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“…Samples were centrifuged at 250 × g, and supernatants were cryopreserved at -80°C. Before removal from BSL3 containment, ETA supernatants were UV-inactivated following our published protocols (44).…”

Section: Sample Processingmentioning

confidence: 99%

“…Stained beads were washed with PBS-BSA and resuspended in 1X PBS. Samples were fixed in 1:6 final dilution of FACS Lysing solution (BD Biosciences), which we showed inactivates SARS-CoV-2 (44).…”

Section: Flowbeat Assaysmentioning

confidence: 99%

“…The levels of human cytokines in serum, nasal swabs, and ETA were measured using a Meso Scale Discovery U-plex custom multiplex assay kit, as we previously described (1). We measured an additional ten analytes (IFN-α2a, IFN-γ, IL-15, IL-29/IFN-λ1, IP-10, MCP-1, MDC, MIP-3β, SDF-1a, and TRAIL) following the manufacturer's protocol (Meso Scale Discovery) in plasma, nasal swabs, and ETA, which were prepared following our established UVC-inactivation protocol (44).…”

Section: Cytokine Quantificationmentioning

confidence: 99%

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Transient anti-interferon autoantibodies in the airways are associated with efficient recovery from COVID-19

Babcock,

Kosters,

Eddins

et al. 2024

Preprint

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Pre-existing anti-interferon alpha (anti-IFN-α) autoantibodies in blood are associated with susceptibility to life-threatening COVID-19. However, it is unclear whether anti-IFN-α autoantibodies in the airways -- the initial site of infection -- can also determine disease outcomes. In this study, we developed a new multiparameter technology, flowBEAT, to quantify and profile the isotypes of anti-IFN-α and anti-SARS-CoV-2 antibodies in longitudinal samples collected over 20 months from the airway and matching blood of 129 donors with mild, moderate, and severe COVID-19. We found unexpectedly that nasal anti-IFN-α autoantibodies were induced post-infection onset in more than 70% of mild to moderate COVID-19 cases and associated with robust anti-SARS-CoV-2 immunity, fewer symptoms, and efficient recovery. Nasal anti-IFN-α autoantibodies followed the peak of host IFN-α production and waned with disease recovery, revealing a regulated balance between IFN-α and anti-IFN-α response. Notably, only a subset of mild to moderate patients progressed to develop systemic anti-IFN-α, which correlated with systemic inflammation and worsened symptoms. In contrast, patients with life-threatening COVID-19 sustained elevated anti-IFN-α in both airways and blood, coupled with uncontrolled viral load and IFN-α production. Our studies thereby reveal a novel protective role for nasal anti-IFN-α autoantibodies in the immunopathology of COVID-19 and, more broadly, suggest that anti-IFN-α may serve an important regulatory function to restore homeostasis following viral invasion of the respiratory mucosa.

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Section: Sample Processingmentioning

confidence: 99%

Section: Flowbeat Assaysmentioning

confidence: 99%

Section: Cytokine Quantificationmentioning

confidence: 99%

See 1 more Smart Citation

Transient anti-interferon autoantibodies in the airways are associated with efficient recovery from COVID-19

Babcock,

Kosters,

Eddins

et al. 2024

Preprint

Self Cite

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Pathogenic neutrophilia drives acute respiratory distress syndrome in severe COVID-19 patients

Eddins

Yang

Kosters

et al. 2021

Preprint

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Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and the ensuing COVID-19 pandemic have caused ~33 million cases and over 585,000 deaths in the United States alone. Troubling disparities in COVID-19-associated mortality emerged early, with nearly 70% of deaths being Black/African-American (AA) patients in some areas, yet targeted studies to delineate features of disease severity within this demographic are scant. Our multi-omics single-cell analyses of immune profiles from airways and matching blood samples of Black/AA patients revealed pronounced pulmonary neutrophilia and low viral burden as a hallmark of severe disease, where neutrophil phenotypes display advanced features of cytokine release syndrome and acute respiratory distress syndrome (ARDS). Cell-cell communication and trajectory analysis reveal a subset of circulating S100A12+/CXCR4+ mature neutrophils that infiltrate the lung via the IL-8/CXCR2 axis. Recruited neutrophils progress toward a transcriptionally active and pathogenic state characterized by exacerbated production of IL-8, IL-1β, IL-6, and CCL3/4 along with elevated levels of neutrophil elastase and myeloperoxidase. Inflammatory monocytes recruited via neutrophil-derived CCL3/4 also produce elevated neutrophil chemotactic factor IL-8, potentiating the sustained neutrophilia in the airways. The IL-8/CXCR2 axis emerges as a potential therapeutic target to reduce pathogenic neutrophilia and constrain ARDS in severe COVID-19.

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Inactivation of SARS Coronavirus 2 and COVID-19 patient samples for contemporary immunology and metabolomics studies

Cited by 2 publications

References 68 publications

Transient anti-interferon autoantibodies in the airways are associated with efficient recovery from COVID-19

Transient anti-interferon autoantibodies in the airways are associated with efficient recovery from COVID-19

Pathogenic neutrophilia drives acute respiratory distress syndrome in severe COVID-19 patients

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