BCR repertoire sequencing: different patterns of B‐cell activation after two Meningococcal vaccines

Abstract

Next-generation sequencing was used to investigate the B cell receptor heavy chain transcript repertoire of different B cell subsets (naïve, marginal zone, IgM memory and IgG memory) at baseline, and of plasma cells 7 days following administration of serogroup ACWY meningococcal polysaccharide and protein-polysaccharide conjugate vaccines. Baseline B cell subsets could be distinguished from each other using a small number of repertoire properties (clonality, mutation from germ-line and complementarity-determining region 3 length) that were conserved between individuals. However, analyzing the complementarity-determining region 3 amino acid sequence (which is particularly important for antigen binding) of the baseline subsets showed few sequences shared between individuals. In contrast, day 7 plasma cells demonstrated nearly tenfold greater sequence sharing between individuals than the baseline subsets, consistent with the plasma cells being induced by the vaccine antigen, and sharing specificity for a more limited range of epitopes. By annotating plasma cell sequences based on IgG subclass usage and mutation, and also comparing them to the sequences of the baseline cell subsets, we were able to identify different signatures after the polysaccharide and conjugate vaccines. Plasma cells produced after conjugate vaccination were predominantly IgG1, and most related to IgG memory cells. In contrast, after polysaccharide vaccination, the plasma cells were predominantly IgG2, less mutated, and were equally likely to be related to marginal zone, IgM memory or IgG memory cells. High-throughput B cell repertoire sequencing thus provides a unique insight into patterns of B cell activation not possible from more conventional measures of immunogenicity.

Investigating the effect of AS03 adjuvant on the plasma cell repertoire following pH1N1 influenza vaccination

“…A total of 19,101,557 raw sequencing reads were obtained, which reduced to 9,505,320 following sequence quality control and filtering steps. Sequences from each sample were clustered using a previously described method to group together sequences that are derived from the same clonal lineage, and also those that may have arisen due to PCR and sequencing error1519. On average, 2,353 (range: 627–4,528) clusters were generated for each sample.…”

“…The converse was true for clusters annotated as specific to pH1N1. The pH1N1 and seasonal influenza-specific sequences were also compared to BCR sequence datasets of plasma cells collected following both Hepatitis B 2026 and Meningococcal ACWY15 vaccination, but no matches were found (Fig. 3a,b).…”

“…An increased understanding of the repertoire of plasma cells produced in response to vaccination could potentially be gained by sequencing their B cell receptor (BCR) heavy chain variable regions1415. Knowing the exact nucleotide sequences allows determination of mutation numbers, which can be used to distinguish between the plasma cells activated from naïve B cells vs. pre-existing memory B cells.…”

Heterogeneity of Memory Marginal Zone B Cells

“…The antibody repertoire expressed by these B cells has been suggested to be selected to bind to carbohydrate, carried by microorganisms, for example. 75 These antigens do not require T cell help for their responses. Furthermore, the heavy chains of the IgM molecules expressed by naïve MZ-B cells contain shorter H-CDR3 regions compared to FO-B cells in rats 53 and mice.…”

Immunoglobulin gene analysis as a tool for investigating human immune responses

“…A diverse response is beneficial, a comparison of Avian flu survivors vs non‐survivors found one of the chief differences was the diversity of the B cell repertoire, where increased diversity correlated with survival 106. Repeated sampling of the repertoire over time can be helpful in identifying potentially protective antibodies107 and convergence of repertoires between different people toward similar Ig genotypes has been shown, for example, in response to influenza,108 meningococcal109, and Dengue110 vaccines. However, finding a convergent signature for equivalent challenging antigen preparations may not always be possible, even when temporal data for the response is available 107.…”