Combinatorial multimer staining and spectral flow cytometry facilitate quantification and characterization of polysaccharide-specific B cell immunity

Hoving, D.; Marques, A. H. C.; Huisman, W.; Nosoh, B. A.; de Kroon, A. C.; van Hengel, O. R. J.; Wu, B. R.; van Helden, P. M.; Urban, B. C.; Dhar, N.; Ferreira, D. M.; Kwatra, G.; Hokke, C. H.; Jochems, S. P.

doi:10.1101/2023.03.07.531507

bioRxiv · 10.1101/2023.03.07.531507

Combinatorial multimer staining and spectral flow cytometry facilitate quantification and characterization of polysaccharide-specific B cell immunity

Hoving, D.·Marques, A. H. C.·Huisman, W.·Nosoh, B. A.·de Kroon, A. C.·van Hengel, O. R. J.·Wu, B. R.·van Helden, P. M.·Urban, B. C.·Dhar, N.·Ferreira, D. M.·Kwatra, G.·Hokke, C. H.·Jochems, S. P.

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Abstract

Bacterial capsular polysaccharides are important vaccine immunogens. However, the study of polysaccharide-specific immune responses has been hindered by technical restrictions. Here, we developed and validated a high-throughput method to analyse antigen-specific B cells using combinatorial staining with fluorescently-labelled capsular polysaccharide multimers. Concurrent staining of 25+ cellular markers further enables the in-depth characterization of polysaccharide-specific cells. We used this assay to simultaneously analyse 14 Streptococcus pneumoniae or 5 Streptococcus agalactiae serotype-specific B cell populations. The phenotype of polysaccharide-specific B cells was associated with serotype specificity, vaccination history and donor population. For example, we observed a link between non-class switched (IgM+) memory B cells and vaccine-inefficient S. pneumoniae serotypes 1 and 3. Moreover, B cells had increased activation in donors from South Africa, which has high-incidence of S. agalactiae invasive disease, compared to European donors. This assay allows for the characterization of heterogeneity in B cell immunity that may underlie immunization efficacy.

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Hoving, D., Marques, A. H. C., Huisman, W., Nosoh, B. A., de Kroon, A. C., van Hengel, O. R. J., Wu, B. R., van Helden, P. M., Urban, B. C., Dhar, N., Ferreira, D. M., Kwatra, G., Hokke, C. H., Jochems, S. P.. 2023-03-10. Combinatorial multimer staining and spectral flow cytometry facilitate quantification and characterization of polysaccharide-specific B cell immunity. https://doi.org/10.1101/2023.03.07.531507

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