@article{1688366cc66241fa931d368f09ab0de6,
title = "B cells expressing IgM B cell receptors of HIV-1 neutralizing antibodies discriminate antigen affinities by sensing binding association rates",
abstract = "HIV-1 envelope (Env) proteins designed to induce neutralizing antibody responses allow study of the role of affinities (equilibrium dissociation constant [KD]) and kinetic rates (association/dissociation rates) on B cell antigen recognition. It is unclear whether affinity discrimination during B cell activation is based solely on Env protein binding KD and whether B cells discriminate among proteins of similar affinities that bind with different kinetic rates. Here, we use a panel of Env proteins and Ramos B cell lines expressing immunoglobulin M (IgM) B cell receptors (BCRs) with specificity for CD4-binding-site broadly neutralizing antibodies to study the role of antigen binding kinetic rates on both early (proximal/distal signaling) and late events (BCR/antigen internalization) in B cell activation. Our results support a kinetic model for B cell activation in which Env protein affinity discrimination is based not on overall KD but on sensing of association rate and a threshold antigen-BCR half-life.",
keywords = "B cell, BCR, CP: Immunology, HIV-1, IgM, affinity, antigen internalization, association rate, calcium mobilization, envelope, phospho-signaling",
author = "Hossain, \{Md Alamgir\} and Kara Anasti and Brian Watts and Kenneth Cronin and Ronald Derking and Bettina Groschel and Kane, \{Advaiti Pai\} and Edwards, \{R. J.\} and David Easterhoff and Jinsong Zhang and Wes Rountree and Yaneth Ortiz and Kevin Saunders and Schief, \{William R.\} and Sanders, \{Rogier W.\} and Laurent Verkoczy and Michael Reth and Alam, \{S. Munir\}",
note = "Funding Information: We thank Dr. Bart Haynes, DHVI, Director of CHAVD (Duke Consortia), for providing facility resources, scientific advice, and critical comments. We are grateful to Rogier W. Sanders, Ronald Derking, and Tom Bijl (Amsterdam UMC) for the expression and production of GT1.2 gp140 trimer and GT1.2 NPs and William Schief, Bettina Groschel, Saman Eskandarzadeh, Yumiko Adachi, Mike Kubitz, Ryan Tingle, and Nicole Phelps at Scripps Research for producing eODGT6 and eODGT8 proteins and eODGT8 60-mer NPs. We thank Kevin Saunders and Elizabeth Donahue (Duke HVI) for expressing and purifying Env gp120 and gp140 trimers and NPs. We thank the DHVI teams from BIAcore Facility, Flow cytometry, Protein Expression (Kevin Saunders) Center, and the DHVI Finance and administrative teams for their support. Research reported in this publication was supported by the National Institute of Allergy and Infectious Diseases (NIAID) of the National Institutes of Health (NIH) under award number R01AI145656 (principal investigator [PI]: S.M.A.). The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. S.M.A. conceived and designed the study and wrote the paper. M.A.H. and K.A. co-wrote and edited the paper. M.R. and L.V. provided collaborative support and revised and edited paper. K.A. performed and analyzed SPR and Ca-flux experiments. M.A.H. phenotyped Ramos cell lines and performed and analyzed cell-surface binding, phospho-signaling, and BCR/antigen downmodulation and internalization experiments. B.W. performed and analyzed thermodynamics experiments. K.C. prepared and purified Fab-trimer complexes. R.J.E. performed and analyzed NSEM data. A.P.K. performed and analyzed antigenicity of Env proteins. J.Z. performed ex vivo Ca-flux experiments. M.R. and Y.O. assisted with the protocols for BCR/antigen downmodulation and internalization. D.E. developed cell lines. M.A.H. phenotyped and sorted for high BCR-expressing Ramos cell lines expressing CH31, VRC01, or VRC01 UCA-IgM BCRs. W.R. performed statistical analysis for correlation evaluation. R.W.S. and R.D. designed and produced GT1.2 gp140 trimers and GT1.2 NPs. W.R.S. and B.G. designed and produced eODGT6 and eODGT8 monomers and 60-mer NPs. All authors reviewed and edited the manuscript. None of the authors have a conflict of interest. Funding Information: We thank Dr. Bart Haynes, DHVI, Director of CHAVD (Duke Consortia), for providing facility resources, scientific advice, and critical comments. We are grateful to Rogier W. Sanders, Ronald Derking, and Tom Bijl (Amsterdam UMC) for the expression and production of GT1.2 gp140 trimer and GT1.2 NPs and William Schief, Bettina Groschel, Saman Eskandarzadeh, Yumiko Adachi, Mike Kubitz, Ryan Tingle, and Nicole Phelps at Scripps Research for producing eODGT6 and eODGT8 proteins and eODGT8 60-mer NPs. We thank Kevin Saunders and Elizabeth Donahue (Duke HVI) for expressing and purifying Env gp120 and gp140 trimers and NPs. We thank the DHVI teams from BIAcore Facility, Flow cytometry, Protein Expression (Kevin Saunders) Center, and the DHVI Finance and administrative teams for their support. Research reported in this publication was supported by the National Institute of Allergy and Infectious Diseases (NIAID) of the National Institutes of Health (NIH) under award number R01AI145656 (principal investigator [PI]: S.M.A.). The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. Publisher Copyright: {\textcopyright} 2022 The Authors",
year = "2022",
month = jun,
day = "28",
doi = "10.1016/j.celrep.2022.111021",
language = "English",
volume = "39",
pages = "111021",
journal = "Cell reports",
issn = "2211-1247",
publisher = "Cell Press",
number = "13",
}
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