Abstract
Vaccination against SARS-CoV-2 with coronavirus vaccines that elicit protective immune responses is critical to the prevention of severe disease and mortality associated with SARS-CoV-2 infection. Understanding the adaptive immune responses to SARS-CoV-2 infection and/or vaccination will continue to aid in the development of next-generation vaccines. Studies have shown the important role of SARS-CoV-2-specific antibodies for both disease resolution and prevention of COVID-19 serious sequelae following vaccination. However, antibody responses are short-lived, highlighting the importance of studying antigen-specific B-cell responses to better understand durable immunity and immunologic memory. Since the spike protein is the main target of antibody-producing B cells, we developed a SARS-CoV-2 memory B cell ELISPOT assay to measure the frequencies of spike-specific B cells after COVID-19 infection and/or vaccination. Here, we describe in detail the methodology for using this ELISPOT assay to quantify SARS-CoV-2 spike-specific memory B cells produced by infection and/or vaccination in human PBMC samples. Application of this assay may help better understand and predict SARS-CoV-2 recall immune responses and to develop potential B cell correlates of protection at the methodological level.
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Acknowledgments
We would like to thank the Mayo Clinic Vaccine Research Group personnel for their technical assistance during the development and execution of these assays.
Conflict of Interest Statement
Dr. Ovsyannikova holds patents related to vaccinia and measles peptide vaccines. Drs. Kennedy and Ovsyannikova hold a patent related to vaccinia peptide vaccines. Drs. Kennedy and Ovsyannikova have received grant funding and royalties from ICW Ventures for preclinical studies on a peptide-based COVID-19 vaccine. Drs. Kennedy, Ovsyannikova, and Haralambieva hold a patent related to the impact of single nucleotide polymorphisms on measles vaccine immunity. Dr. Kennedy has received funding from Merck Research Laboratories to study waning immunity to mumps vaccine. Dr. Kennedy also offers consultative advice on vaccine development to Merck & Co. and Sanofi Pasteur. These activities have been reviewed by the Mayo Clinic Conflict of Interest Review Board and are conducted in compliance with the Mayo Clinic Conflict of Interest policies. This research has been reviewed by the Mayo Clinic Conflict of Interest Review Board and was conducted in compliance with the Mayo Clinic Conflict of Interest policy.
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Swanson, I.M., Haralambieva, I.H., Rasche, M.M., Ovsyannikova, I.G., Kennedy, R.B. (2024). Frequencies of SARS-CoV-2 Spike Protein-Specific Memory B Cells in Human PBMCs, Quantified by ELISPOT Assay. In: Kalyuzhny, A.E. (eds) Handbook of ELISPOT . Methods in Molecular Biology, vol 2768. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3690-9_10
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DOI: https://doi.org/10.1007/978-1-0716-3690-9_10
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