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

Hoving, Dennis, Marques, Alexandre H. C., Huisman, Wesley, Nosoh, Beckley A., de Kroon, Alicia C., van Hengel, Oscar R. J., Wu, Bing-Ru, Steenbergen, Rosanne A. M., van Helden, Pauline M., Urban, Britta ORCID: https://orcid.org/0000-0002-4197-8393, Dhar, Nisha, Ferreira, Daniela ORCID: https://orcid.org/0000-0002-0594-0902, Kwatra, Gaurav, Hokke, Cornelis H. and Jochems, Simon P. (2023) 'Combinatorial multimer staining and spectral flow cytometry facilitate quantification and characterization of polysaccharide-specific B cell immunity'. Communications Biology, Vol 6, Issue 1, e1095.

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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 Dutch donors. This assay allows for the characterization of heterogeneity in B cell immunity that may underlie immunization efficacy.

Item Type:	Article
Subjects:	QW Microbiology and Immunology > Immunotherapy and Hypersensitivity > QW 806 Vaccination
Faculty: Department:	Clinical Sciences & International Health > Clinical Sciences Department
Digital Object Identifer (DOI):	https://doi.org/10.1038/s42003-023-05444-3
SWORD Depositor:	JISC Pubrouter
Depositing User:	JISC Pubrouter
Date Deposited:	30 Oct 2023 08:02
Last Modified:	30 Oct 2023 08:10
URI:	https://archive.lstmed.ac.uk/id/eprint/23379

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