Abstract
Immune response following subsequent encounter of viruses (and vaccines) relies largely on the pool and frequencies of antigen-specific memory B cells. In addition to antibody titers, the reliable measurement of these cells in human blood (peripheral blood mononuclear cells/PBMCs or purified B cells) provides valuable information on the preparedness of the adaptive immune system to respond to infection or vaccines, and potentially supports the discovery of new quantitative correlates of protection. The Mayo Clinic Vaccine Research Group has developed and optimized a high-throughput ELISPOT-based assay for the quantification of influenza A/H1N1 virus-specific memory B cells in human PBMCs. Here, we present the materials and detailed methodology for using this assay on cryopreserved cells for the measurement of recall humoral immunity readiness (antigen-specific memory B cell frequencies) after influenza vaccination. This assay could be readily adapted to other influenza virus strains and other respiratory viruses and vaccines for use in systems biology and larger population-based studies.
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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. We also thank Caroline L. Vitse for her editorial assistance. This work was supported by the National Institute of Allergy And Infectious Diseases of the National Institutes of Health under award number U01AI089859.
Dr. Poland is the chair of a Safety Evaluation Committee for novel investigational vaccine trials being conducted by Merck Research Laboratories. Dr. Poland offers consultative advice on vaccine development to Merck & Co. Inc., Avianax, Adjuvance, Alopexx, Sanofi Pasteur, GlaxoSmithKline, and Emergent Biosolutions. Drs. Poland and Ovsyannikova hold four patents related to vaccinia and measles peptide research. Dr. Kennedy has received funding from Merck Research Laboratories to study waning immunity to mumps vaccine. These activities have been reviewed by the Mayo Clinic Conflict of Interest Review Board and are conducted in compliance with 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 Mayo Clinic Conflict of Interest policies.
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Haralambieva, I.H., Ovsyannikova, I.G., Kennedy, R.B., Poland, G.A. (2018). Detection and Quantification of Influenza A/H1N1 Virus-Specific Memory B Cells in Human PBMCs Using ELISpot Assay. In: Kalyuzhny, A. (eds) Handbook of ELISPOT . Methods in Molecular Biology, vol 1808. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8567-8_19
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DOI: https://doi.org/10.1007/978-1-4939-8567-8_19
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