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

bioRxiv (Cold Spring Harbor Laboratory)

Abstract

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.

Authors 14

1. Dennis Hoving corresponding Aachen

   Leiden University · Leiden University Medical Center

   Affiliation as printed

   Leiden University Center for Infectious Diseases, Leiden University Medical Center, Leiden, the Netherlands

2. A.H.C. Marques Aachen

   Leiden University · Leiden University Medical Center

   Affiliation as printed

   Leiden University Center for Infectious Diseases, Leiden University Medical Center, Leiden, the Netherlands

3. Wesley Huisman Aachen

   Leiden University · Leiden University Medical Center

   Affiliation as printed

   Leiden University Center for Infectious Diseases, Leiden University Medical Center, Leiden, the Netherlands

4. Beckley A. Nosoh Aachen

   Leiden University · Leiden University Medical Center

   Affiliation as printed

   Leiden University Center for Infectious Diseases, Leiden University Medical Center, Leiden, the Netherlands

5. Alicia C. de Kroon Aachen

   Leiden University · Leiden University Medical Center

   Affiliation as printed

   Leiden University Center for Infectious Diseases, Leiden University Medical Center, Leiden, the Netherlands

6. Oscar R. J. van Hengel Aachen

   Leiden University · Leiden University Medical Center

   Affiliation as printed

   Leiden University Center for Infectious Diseases, Leiden University Medical Center, Leiden, the Netherlands

7. B.R. Wu Aachen

   Leiden University · Leiden University Medical Center

   Affiliation as printed

   Leiden University Center for Infectious Diseases, Leiden University Medical Center, Leiden, the Netherlands

8. Pauline M. van Helden

   AIMM Therapeutics (Netherlands)

   Affiliation as printed

   AIMM Therapeutics, Amsterdam, the Netherlands. Current address: LAVA Therapeutics N.V., Utrecht, the Netherlands

9. Britta C. Urban

   Liverpool School of Tropical Medicine · University of Oxford

   Affiliation as printed

   Department of Clinical Sciences, Liverpool School of Tropical Medicine, Liverpool, UK

   Oxford Vaccine Group, University of Oxford, Oxford, UK

10. Nisha Dhar

    University of the Witwatersrand

    Affiliation as printed

    Vaccines & Infectious Diseases Analytics, University of Witwatersrand, Johannesburg, South Africa

11. Daniela Mulari Ferreira

    Liverpool School of Tropical Medicine · University of Oxford

    Affiliation as printed

    Department of Clinical Sciences, Liverpool School of Tropical Medicine, Liverpool, UK

    Oxford Vaccine Group, University of Oxford, Oxford, UK

12. Gaurav Kwatra

    Christian Medical College, Vellore · University of the Witwatersrand

    Affiliation as printed

    Department of Clinical Microbiology, Christian Medical College, Vellore, India

    Vaccines & Infectious Diseases Analytics, University of Witwatersrand, Johannesburg, South Africa

13. Cornelis Hendrik Hokke Aachen

    Leiden University · Leiden University Medical Center

    Affiliation as printed

    Leiden University Center for Infectious Diseases, Leiden University Medical Center, Leiden, the Netherlands

14. Simon P. Jochems corresponding Aachen

    Leiden University · Leiden University Medical Center

    Affiliation as printed

    Leiden University Center for Infectious Diseases, Leiden University Medical Center, Leiden, the Netherlands