Transcriptional and clonal characterization of B cell plasmablast diversity following primary and secondary natural DENV infection
Adam T. Waickman,
Gregory D. Gromowski,
Wiriya Rutvisuttinunt,
Tao Li,
Hayden Siegfried,
Kaitlin Victor,
Caitlin Kuklis,
Methee Gomootsukavadee,
Michael K. McCracken,
Benjamin Gabriel,
Anuja Mathew,
Ariadna Grinyo i Escuer,
Mallorie E. Fouch,
Jenny Liang,
Stefan Fernandez,
Edgar Davidson,
Benjamin J. Doranz,
Anon Srikiatkhachorn,
Timothy Endy,
Stephen J. Thomas,
Damon Ellison,
Alan L. Rothman,
Richard G. Jarman,
Jeffrey R. Currier,
Heather Friberg
Affiliations
Adam T. Waickman
Viral Diseases Branch, Walter Reed Army Institute of Research, Silver Spring, MD, United States; Corresponding author.
Gregory D. Gromowski
Viral Diseases Branch, Walter Reed Army Institute of Research, Silver Spring, MD, United States
Wiriya Rutvisuttinunt
Viral Diseases Branch, Walter Reed Army Institute of Research, Silver Spring, MD, United States
Tao Li
Viral Diseases Branch, Walter Reed Army Institute of Research, Silver Spring, MD, United States
Hayden Siegfried
Viral Diseases Branch, Walter Reed Army Institute of Research, Silver Spring, MD, United States
Kaitlin Victor
Viral Diseases Branch, Walter Reed Army Institute of Research, Silver Spring, MD, United States
Caitlin Kuklis
Viral Diseases Branch, Walter Reed Army Institute of Research, Silver Spring, MD, United States
Methee Gomootsukavadee
Department of Virology, Armed Forces Research Institute of Medical Sciences, Bangkok, Thailand
Michael K. McCracken
Viral Diseases Branch, Walter Reed Army Institute of Research, Silver Spring, MD, United States
Benjamin Gabriel
Department of Cell and Molecular Biology, Institute for Immunology and Informatics, University of Rhode Island, Providence, RI, United States
Anuja Mathew
Department of Cell and Molecular Biology, Institute for Immunology and Informatics, University of Rhode Island, Providence, RI, United States
Ariadna Grinyo i Escuer
Integral Molecular, Philadelphia, PA, United States
Mallorie E. Fouch
Integral Molecular, Philadelphia, PA, United States
Jenny Liang
Integral Molecular, Philadelphia, PA, United States
Stefan Fernandez
Department of Virology, Armed Forces Research Institute of Medical Sciences, Bangkok, Thailand
Edgar Davidson
Integral Molecular, Philadelphia, PA, United States
Benjamin J. Doranz
Integral Molecular, Philadelphia, PA, United States
Anon Srikiatkhachorn
Department of Cell and Molecular Biology, Institute for Immunology and Informatics, University of Rhode Island, Providence, RI, United States; Faculty of Medicine, King Mongkut's Institute of Technology Ladkrabang, Bangkok, Thailand
Timothy Endy
Department of Microbiology and Immunology, State University of New York Upstate Medical University, Syracuse, New York, USA
Stephen J. Thomas
Institute for Global Health and Translational Sciences, State University of New York Upstate Medical University, Syracuse, New York, USA.
Damon Ellison
Viral Diseases Branch, Walter Reed Army Institute of Research, Silver Spring, MD, United States
Alan L. Rothman
Department of Cell and Molecular Biology, Institute for Immunology and Informatics, University of Rhode Island, Providence, RI, United States
Richard G. Jarman
Viral Diseases Branch, Walter Reed Army Institute of Research, Silver Spring, MD, United States
Jeffrey R. Currier
Viral Diseases Branch, Walter Reed Army Institute of Research, Silver Spring, MD, United States
Heather Friberg
Viral Diseases Branch, Walter Reed Army Institute of Research, Silver Spring, MD, United States
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. Keywords: Dengue, Plasmablast, Single-cell RNA sequencing, IgA, Monoclonal antibody
