Department of Medicine
Immunoprophylaxis and Therapy | Therapeutics
In the current study, an improved NGS approach was developed to study the B-cell repertoire evolution in a simple mouse immunization model including only two DNA immunizations. The combination of 5'RACE and Ion Torrent long reads enabled unbiased immunoglobulin repertoire analysis even from small amounts of peripheral mouse blood. The B-cell population expanded by the vaccine displayed a relatively strong clonality. Upon priming with the first vaccine dose, we observed a consistent pattern of V-segment gene and CDR3 usage (public specificities). Interestingly, this pattern diversified with the second dose of immunization -it was relatively different in individual mice in spite of having received the same vaccine regimen (private specificities). Nevertheless, there were several instances in which the same public V-segment genes and CDR3s that were expanded after the first dose were further amplified after the second immunization. Taken together, it appears that the major clonotypes expanded by vaccination were originally a homogeneous subset that later diversified after a second dose leading to diverse "private" clonal compositions in different mice. These results established a new platform valuable to perform longitudinal analyses of the Ig germline gene usage and clonotype evolution throughout an immunization regimen in a commonly used animal model.
DNA vaccine, germline, Immunoglobulin, Next-generation sequencing, V-gene, vaccine
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Copyright © 2017 The Author(s). Published with license by Taylor and Francis. This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivatives License (http://creativecommons.org/licenses/by-nc-nd/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited, and is not altered, transformed, or built upon in any way.
DOI of Published Version
Hum Vaccin Immunother. 2017 Dec 2;13(12):2987-2995. doi: 10.1080/21645515.2017.1379638. Epub 2017 Oct 19. Link to article on publisher's site
Human vaccines and immunotherapeutics
Farfán Arribas DJ, Liu S, Wang S, Lu S. (2017). The dynamics of immunoglobulin V-gene usage and clonotype expansion in mice after prime and boost immunizations as analyzed by NGS. Open Access Articles. https://doi.org/10.1080/21645515.2017.1379638. Retrieved from https://escholarship.umassmed.edu/oapubs/3276
Creative Commons License
This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 4.0 License.