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Development of flow cytometry-based assays to assess the ability of antibodies to bind to SARS-CoV-2-infected and spike-transfected cells and mediate NK cell degranulation.
Mielke, Dieter; Stanfield-Oakley, Sherry; Jha, Shalini; Keyes, Taylor; Zalaquett, Adam; Dunn, Brooke; Rodgers, Nicole; Oguin, Thomas; Sempowski, Greg D; Binder, Raquel A; Gray, Gregory C; Karuna, Shelly; Corey, Lawrence; Hural, John; Tomaras, Georgia D; Pollara, Justin; Ferrari, Guido.
  • Mielke D; Department of Surgery, Duke University, Durham, North Carolina, USA.
  • Stanfield-Oakley S; Department of Surgery, Duke University, Durham, North Carolina, USA.
  • Jha S; Department of Surgery, Duke University, Durham, North Carolina, USA.
  • Keyes T; Department of Surgery, Duke University, Durham, North Carolina, USA.
  • Zalaquett A; Department of Surgery, Duke University, Durham, North Carolina, USA.
  • Dunn B; Department of Surgery, Duke University, Durham, North Carolina, USA.
  • Rodgers N; Department of Surgery, Duke University, Durham, North Carolina, USA.
  • Oguin T; Duke Human Vaccine Institute, Duke University School of Medicine, Durham, North Carolina, USA.
  • Sempowski GD; Duke Human Vaccine Institute, Duke University School of Medicine, Durham, North Carolina, USA.
  • Binder RA; Department of Pathology, Duke University, Durham, North Carolina, USA.
  • Gray GC; Department of Medicine, Duke University, Durham, North Carolina, USA.
  • Karuna S; Duke Global Health Institute, Duke University, Durham, North Carolina, USA.
  • Corey L; Division of Infectious Diseases, Duke University, Durham, North Carolina, USA.
  • Hural J; Vaccine Infectious Disease Division, Fred Hutchinson Cancer Center, Seattle, Washington, USA.
  • Tomaras GD; Vaccine Infectious Disease Division, Fred Hutchinson Cancer Center, Seattle, Washington, USA.
  • Pollara J; Department of Medicine, University of Washington, Seattle, Washington, USA.
  • Ferrari G; Department of Laboratory Medicine and Pathology, University of Washington, Seattle, Washington, USA.
Cytometry A ; 101(6): 483-496, 2022 06.
Article in English | MEDLINE | ID: covidwho-1750349
ABSTRACT
Since the beginning of the SARS-CoV-2 pandemic, antibody responses and antibody effector functions targeting SARS-CoV-2-infected cells have been understudied. Consequently, the role of these types of antibodies in SARS-CoV-2 disease (COVID-19) and immunity is still undetermined. To provide tools to study these responses, we used plasma from SARS-CoV-2-infected individuals (n = 50) and SARS-CoV-2 naive healthy controls (n = 20) to develop four specific and reproducible flow cytometry-based assays (i) two assessing antibody binding to, and antibody-mediated NK cell degranulation against, SARS-CoV-2-infected cells and (ii) two assessing antibody binding to, and antibody-mediated NK cell degranulation against, SARS-CoV-2 Spike-transfected cells. All four assays demonstrated the ability to detect the presence of these functional antibody responses in a specific and reproducible manner. Interestingly, we found weak to moderate correlations between the four assays (Spearman rho ranged from 0.50 to 0.74), suggesting limited overlap in the responses captured by the individual assays. Lastly, while we initially developed each assay with multiple dilutions in an effort to capture the full relationship between antibody titers and assay outcome, we explored the relationship between fewer antibody dilutions and the full dilution series for each assay to reduce assay costs and improve assay efficiency. We found high correlations between the full dilution series and fewer or single dilutions of plasma. Use of single or fewer sample dilutions to accurately determine the response rates and magnitudes of the responses allows for high-throughput use of these assays platforms to facilitate assessment of antibody responses elicited by SARS-CoV-2 infection and vaccination in large clinical studies.
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Full text: Available Collection: International databases Database: MEDLINE Main subject: SARS-CoV-2 / COVID-19 Type of study: Prognostic study Topics: Vaccines Limits: Humans Language: English Journal: Cytometry A Year: 2022 Document Type: Article Affiliation country: Cyto.a.24552

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Full text: Available Collection: International databases Database: MEDLINE Main subject: SARS-CoV-2 / COVID-19 Type of study: Prognostic study Topics: Vaccines Limits: Humans Language: English Journal: Cytometry A Year: 2022 Document Type: Article Affiliation country: Cyto.a.24552