| Title: |
A Comparison of Flow Cytometry-based versus ImmunoSpot- or Supernatant-based Detection of SARS-CoV-2 Spike-specific Memory B Cells in Peripheral Blood. |
| Authors: |
Stylianou, Georgia; Cookson, Sharon; Nassif, Justin T.; Kirchenbaum, Greg A.; Lehmann, Paul V.; Todryk, Stephen M. |
| Source: |
Vaccines; Jan2026, Vol. 14 Issue 1, p20, 15p |
| Subject Terms: |
Flow cytometry; SARS-CoV-2; Immunoassay; Mononuclear leukocytes; Antibody formation; Immunologic memory; Detection algorithms |
| Abstract: |
Background: Memory B cells (Bmem) facilitate the generation of renewed and rapid antigen-specific antibody responses long after the initial antigen exposure, at a time when circulating serum antibodies may have declined. As the generation and/or recruitment of Bmem is at the core of most vaccination strategies, the assessment of antigen-specific Bmem is highly informative for forecasting and profiling the elicited B cell immune response. Methods: The two prevalent techniques used to detect antigen-specific Bmem cells at single-cell resolution are probe-based flow cytometry and B cell ImmunoSpot, while the measurement of B cell-derived antibodies in culture supernatants of stimulated B cells offers a semi-quantitative alternative. To the best of our knowledge, a direct side-by-side comparison of these assay systems has not yet been reported using the same starting PBMC material in a blinded fashion to test all three assays simultaneously. Results: These three assay systems were run in parallel to detect SARS-CoV-2 Wuhan-1 strain Spike-specific IgG+ Bmem in peripheral blood mononuclear cell (PBMC) samples obtained from well-defined cohorts comprising pre-COVID-19 era "naïve" individuals (negative controls), individuals shortly after recovery from a PCR-verified SARS-CoV-2 infection (positive controls), and a cohort of donor PBMCs isolated in 2024 (the experimental group). Each assay was able to discern Spike-exposed individuals from naïve , with ImmunoSpot suggesting superior sensitivity and specificity. ImmunoSpot and flow cytometry results were closely correlated. Conclusions: The study demonstrates that all three assays are suited for the detection of specific Bmem in antigen-primed individuals when such Bmem occur in the mid- to high-frequency range, and that they broadly concur. Strengths and weaknesses of the three test systems are discussed. [ABSTRACT FROM AUTHOR] |
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| Database: |
Complementary Index |