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

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<sup>+</sup>) 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 Dutch donors. This assay allows for the characterization of heterogeneity in B cell immunity that may underlie immunization efficacy.
Publication status:
Published
Peer review status:
Peer reviewed

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Publisher copy:
10.1038/s42003-023-05444-3

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Funder identifier:
https://ror.org/0456r8d26


Publisher:
Springer Nature
Journal:
Communications Biology More from this journal
Volume:
6
Issue:
1
Article number:
1095
Place of publication:
England
Publication date:
2023-10-28
Acceptance date:
2023-10-11
DOI:
EISSN:
2399-3642
ISSN:
2399-3642
Pmid:
37898698

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