title: Single-cell and deep sequencing of IgG-switched macaque B cells reveal a diverse Ig repertoire following immunization.
abstract: The nonhuman primate model is important for preclinical evaluation of prophylactic and therapeutic intervention strategies. The recent description of the rhesus macaque germline Ig loci and establishment of a database of germline gene segments offer improved opportunities to delineate Ig gene usage in the overall B cell repertoire as well as in response to vaccination. We applied 454-pyrosequencing and single-cell RT-PCR of bulk and sorted memory B cells, respectively, to investigate IGHV gene segment expression in rhesus macaques. The two methods gave remarkably concordant results and identified groups of gene segments that are frequently or rarely used. We further examined the VH repertoire of Ag-specific memory B cells induced by immunization with recombinant HIV-1 envelope glycoproteins, an important vaccine component. We demonstrate that HIV-1 envelope glycoprotein immunization activates a highly polyclonal response composed of most of the expressed VH gene segments, illustrating the considerable genetic diversity of responding B cells following vaccination.
authors.affiliation: Department of Microbiology, Tumor and Cell Biology, Karolinska Institutet, SE-171 77 Stockholm, Sweden;
authors.forename: Christopher
authors.initials: C
authors.lastname: Sundling
authors.forename: Zhenhai
authors.initials: Z
authors.lastname: Zhang
authors.forename: Ganesh E
authors.initials: GE
authors.lastname: Phad
authors.forename: Zizhang
authors.initials: Z
authors.lastname: Sheng
authors.forename: Yimeng
authors.initials: Y
authors.lastname: Wang
authors.forename: John R
authors.initials: JR
authors.lastname: Mascola
authors.forename: Yuxing
authors.initials: Y
authors.lastname: Li
authors.forename: Richard T
authors.initials: RT
authors.lastname: Wyatt
authors.forename: Lawrence
authors.initials: L
authors.lastname: Shapiro
authors.forename: Gunilla B
authors.initials: GB
authors.lastname: Karlsson Hedestam
mesh_terms: D000818:Animals; D001402:B-Lymphocytes / Q000276:immunology* / Q000378:metabolism*; D015483:HIV Antibodies / Q000235:genetics / Q000276:immunology; D015497:HIV-1 / Q000276:immunology; D059014:High-Throughput Nucleotide Sequencing; D007114:Immunization; D017578:Immunoglobulin Class Switching; D007074:Immunoglobulin G / Q000235:genetics*; D007143:Immunoglobulin Heavy Chains / Q000235:genetics; D007135:Immunoglobulin Variable Region / Q000235:genetics; D007136:Immunoglobulins / Q000235:genetics* / Q000276:immunology*; D007156:Immunologic Memory; D008253:Macaca mulatta; D008969:Molecular Sequence Data; D059010:Single-Cell Analysis
chemical_list: D015483:HIV Antibodies; D007074:Immunoglobulin G; D007143:Immunoglobulin Heavy Chains; D007135:Immunoglobulin Variable Region; D007136:Immunoglobulins