Genetic measurement of memory B-cell recall using antibody repertoire sequencing
- 29 July 2013
- journal article
- research article
- Published by Proceedings of the National Academy of Sciences in Proceedings of the National Academy of Sciences
- Vol. 110 (33) , 13463-13468
- https://doi.org/10.1073/pnas.1312146110
Abstract
Annual influenza vaccinations aim to protect against seasonal infections, and vaccine strain compositions are updated every year. This protection is based on antibodies that are produced by either newly activated or memory B cells recalled from previous encounters with influenza vaccination or infection. The extent to which the B-cell repertoire responds to vaccination and recalls antibodies has so far not been analyzed at a genetic level--which is to say, at the level of antibody sequences. Here, we developed a consensus read sequencing approach that incorporates unique barcode labels on each starting RNA molecule. These labels allow one to combine multiple sequencing reads covering the same RNA molecule to reduce the error rate to a desired level, and they also enable accurate quantification of RNA and isotype levels. We validated this approach and analyzed the differential response of the antibody repertoire to live-attenuated or trivalent-inactivated influenza vaccination. Additionally, we analyzed the antibody repertoire in response to repeated yearly vaccinations with trivalent-inactivated influenza vaccination. We found antibody sequences that were present in both years, providing a direct genetic measurement of B-cell recall.Keywords
This publication has 25 references indexed in Scilit:
- Statistical inference of the generation probability of T-cell receptors from sequence repertoiresProceedings of the National Academy of Sciences, 2012
- Pandemic H1N1 influenza vaccine induces a recall response in humans that favors broadly cross-reactive memory B cellsProceedings of the National Academy of Sciences, 2012
- Digital RNA sequencing minimizes sequence-dependent bias and amplification noise with optimized single-molecule barcodesProceedings of the National Academy of Sciences, 2012
- Systems biology of vaccination for seasonal influenza in humansNature Immunology, 2011
- Detection and quantification of rare mutations with massively parallel sequencingProceedings of the National Academy of Sciences, 2011
- Counting individual DNA molecules by the stochastic attachment of diverse labelsProceedings of the National Academy of Sciences, 2011
- Determinism and stochasticity during maturation of the zebrafish antibody repertoireProceedings of the National Academy of Sciences, 2011
- Precise determination of the diversity of a combinatorial antibody library gives insight into the human immunoglobulin repertoireProceedings of the National Academy of Sciences, 2009
- High-Throughput Sequencing of the Zebrafish Antibody RepertoireScience, 2009
- Rapid cloning of high-affinity human monoclonal antibodies against influenza virusNature, 2008