Transcriptional and clonal characterization of B cell plasmablast diversity following primary and secondary natural DENV infection.

Antibody-mediated humoral immunity is thought to play a central role in mediating the immunopathogenesis of acute DENV infection, but limited data are available on the diversity, specificity, and functionality of the antibody response at the molecular level elicited by primary or secondary DENV infection. In order to close this functional gap in our understanding of DENV-specific humoral immunity, we utilized high-throughput single cell RNA sequencing to investigate B cells circulating in both primary and secondary natural DENV infections. We captured full-length paired immunoglobulin receptor sequence data from 9,027 B cells from a total of 6 subjects, including 2,717 plasmablasts. In addition to IgG and IgM class-switched cells, we unexpectedly found a high proportion of the DENV-elicited plasmablasts expressing IgA, principally in individuals with primary DENV infections. These IgA class-switched cells were extensively hypermutated even in individuals with a serologically confirmed primary DENV infection. Utilizing a combination of conventional biochemical assays and high-throughput shotgun mutagenesis, we determined that DENV-reactive IgA class-switched antibodies represent a significant fraction of DENV-reactive Igs generated in response to DENV infection, and that they exhibit a comparable epitope specificity to DENV-reactive IgG antibodies. These results provide insight into the molecular-level diversity of DENV-elicited humoral immunity and identify a heretofore unappreciated IgA plasmablast response to DENV infection.

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Copyright © 2020 The Authors. Published by Elsevier B.V. All rights reserved.

Déclaration de conflit d'intérêts

Declaration of Competing Interest A.T.W reports grants from Military Infectious Disease Research Program, during the conduct of the study. B.J.D, E.D., M.E.F, A.G., J.L., and A.L.R. reports grants from National Institute of Allergy and Infectious Diseases, during the conduct of the study. B.J.D. reports other support from Integral Molecular, outside the submitted work. S.J.T reports other support from US DoD, other support from GSK, during the conduct of the study; personal fees and other support from GSK Vaccines, personal fees and other support from Takeda, personal fees and other support from Merck, personal fees and other support from PrimeVax, personal fees and other support from Themisbio, personal fees and other support from Chugai Pharma, personal fees and other support from Cormac Life Sciences, personal fees and other support from HHS NVPO / Tunnel Govt Services, personal fees and other support from Janssen, other support from GreenMark Partners, personal fees from Tremeau Pharma, outside the submitted work; In addition, Dr. Thomas has a patent US10086061B2 (combined flavivirus vaccines) issued. All other authors have nothing to disclose.