Signatures of the neutralizing dengue virus-reactive memory B cell repertoire identified by multiplexed envelope dimer probes
Year: 2026
Type: preprint
Abstract: Sequential dengue virus (DENV) infection is thought to induce long-lived protection against dengue. However, little is known about the specificity, phenotype, and repertoire of the enduring B cell response due to limitations in high-throughput methods. We used stabilized DENV envelope (E) dimers as probes for in-depth memory B cell repertoire characterization using two complementary sequencing strategies. Using Rapid Assembly, Transfection, and Production of Immunoglobulins (RATP-Ig), we isolated 227 DENV2 E dimer-specific memory B cells from an immune donor, of which 87% bound at least two serotypes, 68% bound the fusion loop (FL-sensitive), and 42% were neutralizing. Among the most neutralizing, cross-binding antibodies, the FL-sensitive antibodies had weaker but broad neutralization, including to other flaviviruses, whereas FL-'agnostic' antibodies mapped to E domain II and III and were potently neutralizing, but only against one or two serotypes. Competitive binding of the DENV1-4 E dimers distinguished the more potently neutralizing FL-agnostic antibodies and broad and potent anti (α)-envelope dimer epitope (EDE) antibodies from the low-quality FL-sensitive antibodies. Further, analysis of the heavy chain complementarity determining region 3 (CDRH3) showed FL-sensitive and α-EDE antibodies were more negatively charged, but α-EDE antibodies were distinguished by longer and more aromatic CDRH3s. We then demonstrated how oligonucleotide-tagged DENV1-4 dimers and proteogenomic single-cell B cell receptor sequencing (LIBRA-seq) could identify memory B cells predicted to have potent neutralization based on DENV-binding and CDRH3 physiochemical properties. This study provides insight into the secondary DENV-specific B cell repertoire and approaches for identifying protective antibodies as part of large-screening efforts.
Source: medRxiv
Authors: Kelsey E. Lowman, Rosemary A. Aogo, Sarah Smith, Ysabelle Broderson, Saba Firdous, Gitanjali Bhushan, Devina J. Thiono, Patrick Mpingabo, Amy R. Henry, Timothy S. Johnston, Jesmine Roberts-Torres, Luis Estrada, Chaim A. Schramm, Foo Cheung, Jessie J. Polanco, Richard Apps, Krista Gangler, Jeffrey I. Cohen, Sonya Jacobson, Edgar Davidson, Benjamin J. Doranz, James Cherry, Iyadh Douagi, Barbara A. Blacklaws, Jonathan L. Heeney, Aravinda M. de Silva, Daniel C. Douek, Leah C. Katzelnick
Language: English
Field: Medicine
Domain: Health Sciences
Open Access status: green
OpenAlex ID: W7213434937