Abstract
A diverse B-cell repertoire is essential for recognition and response to infectious and vaccine antigens. High-throughput sequencing of B-cell receptor (BCR) genes can now be used to study the B-cell repertoire at great depth and may shed more light on B-cell responses than conventional immunological methods. Here, we use high-throughput BCR sequencing to provide novel insight into B-cell dynamics following a primary course of hepatitis B vaccination. Nine vaccine-naive participants were administered three doses of hepatitis B vaccine (months 0, 1, and 2 or 7). High-throughput Illumina sequencing of the total BCR repertoire was combined with targeted sequencing of sorted vaccine antigen-enriched B cells to analyze the longitudinal response of both the total and vaccine-specific repertoire after each vaccine. ELISpot was used to determine vaccine-specific cell numbers following each vaccine. Deconvoluting the vaccine-specific from total BCR repertoire showed that vaccine-specific sequence clusters comprised <0.1 % of total sequence clusters, and had certain stereotypic features. The vaccine-specific BCR sequence clusters were expanded after each of the three vaccine doses, despite no vaccine-specific B cells being detected by ELISpot after the first vaccine dose. These vaccine-specific BCR clusters detected after the first vaccine dose had distinct properties compared to those detected after subsequent doses; they were more mutated, present at low frequency even prior to vaccination, and appeared to be derived from more mature B cells. These results demonstrate the high-sensitivity of our vaccine-specific BCR analysis approach and suggest an alternative view of the B-cell response to novel antigens. In the response to the first vaccine dose, many vaccine-specific BCR clusters appeared to largely derive from previously activated cross-reactive B cells that have low affinity for the vaccine antigen, and subsequent doses were required to yield higher affinity B cells.
Highlights
A diverse B-cell repertoire is essential for recognition and response to infectious and vaccine antigens
We and others have shown that the total repertoire undergoes stereotypic changes following vaccination—the repertoire has an increase in mutation and a decrease in diversity 7 days following vaccination, consistent with an increase in the number of mutated plasma cells (PCs) released into the peripheral blood at this time [2, 5, 6]
Vaccine-specific cells are detected after vaccines 2 and 3 but not after vaccine 1 ELISpot was used to determine the number of HBsAgspecific IgG PCs and memory cells in the peripheral blood at each visit (Fig. 1)
Summary
A diverse B-cell repertoire is essential for recognition and response to infectious and vaccine antigens. High-throughput sequencing of B-cell receptor (BCR) genes can be used to study the B-cell repertoire at great depth and may shed more light on B-cell responses than conventional immunological methods. We use high-throughput BCR sequencing to provide novel insight into B-cell dynamics following a primary course of hepatitis B vaccination. Laserson et al showed that certain clones within the global B-cell repertoire undergo rapid expansions and contractions in response to vaccination [3]. These expansion dynamics were qualitatively different in different individuals and were not related to vaccine type. Focusing on the public repertoire can overcome this to an extent [7], but the public repertoire is enriched for clones specific to antigens that are commonly encountered by the population (e.g., tetanus toxoid, influenza) [10] and it is not clear to what extent the functional properties of the public antigen-specific repertoire and the private antigen-specific repertoire are different
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