Comprehensive baseline characterization of rhesus macaque B cell subtypes and VDJ repertoire via single-cell RNA/VDJ sequencing 2534

Description Rhesus macaques (RMs) are used widely for pre-clinical infection and vaccination studies, yet their B cell immunobiology and immunogenetics remain little characterized. In this study, single-cell RNA/VDJ-sequencing (scRNA/VDJ-seq) was conducted on peripheral blood mononuclear cell (PBMC) samples from 3 male and 3 female, 2-3 years old juvenile RM. From a total 48,888 PBMCs sequenced, 9 RM B cell clusters of distinct transcriptional states were identified, including naïve B cells (∼13% of total B cells), activated B cells (∼7%), unswitched memory B cells (∼36%, 2 subtypes), switched memory B cells (∼23%, 2 subtypes), marginal zone-like B cells (∼19%), natural killer-like B cells (∼1.6%) and antibody-secreting cells (∼0.2%). For each subset, IGH/K/L-V(D)JC gene family and subtype usage, and VDJ repertoire properties such as somatic hypermutation rate and levels, complementarity-determining region heavy chain (CDRH3) amino acid (AA) length and CDRH3 AA hydrophobicity scores were elucidated. While comparable to human B cell subsets, comprehensive analysis of baseline RM PBMC B cell subtypes, paired with their respective V(D)JC repertoire characteristics, reveals distinct properties of RM B cell subsets. Establishment of this library of data will enable a more complete analysis of humoral responses in this critical pre-clinical animal model, providing data for improved predictions of humoral responses observed in RM to anticipated results in humans.