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

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...

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Bibliographic Details
Published in:Communications biology 2023-10, Vol.6 (1), p.1095-12, Article 1095
Main Authors: 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 C., Dhar, Nisha, Ferreira, Daniela M., Kwatra, Gaurav, Hokke, Cornelis H., Jochems, Simon P.
Format: Article
Language:English
Subjects:
631/1647/664
631/250/2152/1566/1571
631/250/2499
631/250/590
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Antigens
Biology
Biomedical and Life Sciences
Capsular polysaccharides
Cell activation
Flow Cytometry
Immunity
Immunization
Immunoglobulin M
Immunological memory
Life Sciences
Lymphocytes B
Memory cells
Phenotypes
Polysaccharides, Bacterial
Serotypes
Streptococcus agalactiae
Streptococcus infections
Streptococcus pneumoniae
Vaccines
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Summary: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. A high-throughput method is developed for simultaneous quantification and in-depth characterization of bacterial polysaccharide antigen-specific B cells.
ISSN:2399-3642
2399-3642
DOI:10.1038/s42003-023-05444-3