Repertoire comparison of the B‐cell receptor‐encoding loci in humans and rhesus macaques by next‐generation sequencing

Abstract

Rhesus macaques (RMs) are a widely used model system for the study of vaccines, infectious diseases and microbial pathogenesis. Their value as a model lies in their close evolutionary relationship to humans, which, in theory, allows them to serve as a close approximation of the human immune system. However, despite their prominence as a human surrogate model system, many aspects of the RM immune system remain ill characterized. In particular, B cell-mediated immunity in macaques has not been sufficiently characterized, and the B-cell receptor-encoding loci have not been thoroughly annotated. To address these gaps, we analyzed the circulating heavy- and light-chain repertoires in humans and RMs by next-generation sequencing. By comparing V gene segment usage, J-segment usage and CDR3 lengths between the two species, we identified several important similarities and differences. These differences were especially notable in the IgM+ B-cell repertoire. However, the class-switched, antigen-educated B-cell populations converged on a set of similar characteristics, implying similarities in how each species responds to antigen. Our study provides the first comprehensive overview of the circulating repertoires of the heavy- and light-chain sequences in RMs, and provides insight into how they may perform as a model system for B cell-mediated immunity in humans.

A method for the isolation and characterization of functional murine monoclonal antibodies by single B cell cloning

“…The light chains were less variable, with CDRL3s of 8–9 amino acids. Both CDR3 length ranges are consistent with antibodies generated in other model systems (34, 35). Each of the six heavy and light chain pairs were co-transfected into HEK293 cells to express recombinant antibodies.…”

Linking EPCR-Binding PfEMP1 to Brain Swelling in Pediatric Cerebral Malaria

“…The initial processing of FASTQ sequence data was performed as previously described (Vigdorovich et al, 2016) with some modifications and additional steps. Briefly, forward and reverse reads were assembled to reconstruct amplicon sequences using FLASH (Magoc and Salzberg, 2011).…”

Germline‐targeting immunogens

“…IOMA is less mutated (22 HC and 15 LC) than VRC01-class antibodies and neutralizes ~50% strains tested (with a mean IC50 of 2.3 μg/ml). IOMA-like bNAbs may thus be elicited more easily than VRC01-class antibodies through immunization, since LCs with 8 AA long CDRL3s are more frequently found in the human repertoire (54, 76, 77), but first, immunogens capable of binding their germline forms must be engineered. e) Although N276-linked carbohydrates are hindering the binding of germline VRC01-class antibodies, they are critically important for the binding of other CD4-BS bNAbs (78, 79).…”

“…Small animals, such as mice, rats, or rabbits (54), as well as non-human primates (NHPs) (77) do not express VH1-2*02 orthologs. In addition, LCs with 5 amino acid long CDRL3s may even be rarer in NHPs (rhesus macaques of Indian origin) than in humans (77).…”

“…IOMA is less mutated (22 HC and 15 LC) than VRC01-class antibodies and neutralizes approximately 50% strains tested (with a mean IC50 of 2.3 μg/mL). IOMA-like bNAbs may thus be elicited more easily than VRC01-class antibodies through immunization, as LCs with eight-amino-acid-long CDRL3s are more frequently found in the human repertoire, 54,77,78 but first, immunogens capable of binding their germline forms must be engineered.…”