Frequency-potency analysis of IgG+ memory B cells delineates neutralizing antibody responses at single-cell resolution
Michelle K. Tenggara,
Seo-Ho Oh,
Catherine Yang,
Hardik K. Nariya,
Amanda M. Metz,
Amit A. Upadhyay,
Dedeepya R. Gudipati,
Lizheng Guo,
Emily G. McGhee,
Kiran Gill,
Elise G. Viox,
Rosemarie D. Mason,
Nicole A. Doria-Rose,
Kathryn E. Foulds,
John R. Mascola,
Yuhong Du,
Haian Fu,
John D. Altman,
Qi Yan,
Zizhang Sheng,
Steven E. Bosinger,
Rui Kong
Affiliations
Michelle K. Tenggara
Emory Vaccine Center, Atlanta, GA 30329, USA; Emory National Primate Research Center, Atlanta, GA 30329, USA
Seo-Ho Oh
Emory Vaccine Center, Atlanta, GA 30329, USA; Emory National Primate Research Center, Atlanta, GA 30329, USA
Catherine Yang
Emory Vaccine Center, Atlanta, GA 30329, USA; Emory National Primate Research Center, Atlanta, GA 30329, USA
Hardik K. Nariya
Emory Vaccine Center, Atlanta, GA 30329, USA; Emory National Primate Research Center, Atlanta, GA 30329, USA
Amanda M. Metz
Emory National Primate Research Center, Atlanta, GA 30329, USA
Amit A. Upadhyay
Emory National Primate Research Center, Atlanta, GA 30329, USA
Dedeepya R. Gudipati
Emory Vaccine Center, Atlanta, GA 30329, USA; Emory National Primate Research Center, Atlanta, GA 30329, USA
Lizheng Guo
Emory Vaccine Center, Atlanta, GA 30329, USA; Emory National Primate Research Center, Atlanta, GA 30329, USA
Emily G. McGhee
Emory Vaccine Center, Atlanta, GA 30329, USA; Emory National Primate Research Center, Atlanta, GA 30329, USA
Kiran Gill
Emory National Primate Research Center, Atlanta, GA 30329, USA
Elise G. Viox
Vaccine Research Center, National Institutes of Health, Bethesda, MD 20892, USA
Rosemarie D. Mason
Vaccine Research Center, National Institutes of Health, Bethesda, MD 20892, USA
Nicole A. Doria-Rose
Vaccine Research Center, National Institutes of Health, Bethesda, MD 20892, USA
Kathryn E. Foulds
Vaccine Research Center, National Institutes of Health, Bethesda, MD 20892, USA
John R. Mascola
Vaccine Research Center, National Institutes of Health, Bethesda, MD 20892, USA
Yuhong Du
Department of Pharmacology and Chemical Biology, Emory University School of Medicine, Atlanta, GA 30322, USA; Emory Chemical Biology Discovery Center, Emory University School of Medicine, Atlanta, GA 30322, USA
Haian Fu
Department of Pharmacology and Chemical Biology, Emory University School of Medicine, Atlanta, GA 30322, USA; Emory Chemical Biology Discovery Center, Emory University School of Medicine, Atlanta, GA 30322, USA; Department of Hematology and Medical Oncology and Winship Cancer Institute, Emory University, Atlanta, GA 30322, USA
John D. Altman
Emory Vaccine Center, Atlanta, GA 30329, USA; Emory National Primate Research Center, Atlanta, GA 30329, USA; Department of Microbiology and Immunology, Emory University School of Medicine, Atlanta, GA 30322, USA
Qi Yan
Department of Obstetrics and Gynecology, Columbia University Irving Medical Center, New York, NY 10032, USA
Zizhang Sheng
Aaron Diamond AIDS Research Center, Columbia University Vagelos College of Physicians and Surgeons, New York, NY 10032, USA
Steven E. Bosinger
Emory Vaccine Center, Atlanta, GA 30329, USA; Emory National Primate Research Center, Atlanta, GA 30329, USA; Department of Pathology and Laboratory Medicine, Emory University School of Medicine, Atlanta, GA 30322, USA
Rui Kong
Emory Vaccine Center, Atlanta, GA 30329, USA; Emory National Primate Research Center, Atlanta, GA 30329, USA; Department of Pathology and Laboratory Medicine, Emory University School of Medicine, Atlanta, GA 30322, USA; Corresponding author
Summary: Identifying individual functional B cell receptors (BCRs) is common, but two-dimensional analysis of B cell frequency versus BCR potency would delineate both quantity and quality of antigen-specific memory B cells. We efficiently determine quantitative BCR neutralizing activities using a single-cell-derived antibody supernatant analysis (SCAN) workflow and develop a frequency-potency algorithm to estimate B cell frequencies at various neutralizing activity or binding affinity cutoffs. In an HIV-1 fusion peptide (FP) immunization study, frequency-potency curves elucidate the quantity and quality of FP-specific immunoglobulin G (IgG)+ memory B cells for different animals, time points, and antibody lineages at single-cell resolution. The BCR neutralizing activities are mainly determined by their affinities to soluble envelope trimer. Frequency analysis definitively demonstrates dominant neutralizing antibody lineages. These findings establish SCAN and frequency-potency analyses as promising approaches for general B cell analysis and monoclonal antibody (mAb) discovery. They also provide specific rationales for HIV-1 FP-directed vaccine optimization.
