title: Antibody repertoire deep sequencing reveals antigen-independent selection in maturing B cells.
abstract: Antibody repertoires are known to be shaped by selection for antigen binding. Unexpectedly, we now show that selection also acts on a non-antigen-binding antibody region: the heavy-chain variable (VH)-encoded "elbow" between variable and constant domains. By sequencing 2.8 million recombined heavy-chain genes from immature and mature B-cell subsets in mice, we demonstrate a striking gradient in VH gene use as pre-B cells mature into follicular and then into marginal zone B cells. Cells whose antibodies use VH genes that encode a more flexible elbow are more likely to mature. This effect is distinct from, and exceeds in magnitude, previously described maturation-associated changes in heavy-chain complementarity determining region 3, a key antigen-binding region, which arise from junctional diversity rather than differential VH gene use. Thus, deep sequencing reveals a previously unidentified mode of B-cell selection.
authors.affiliation: Department of Pathology andDepartment of Systems Biology, Harvard Medical School, Boston, MA 02115; and.
authors.forename: Joseph
authors.initials: J
authors.lastname: Kaplinsky
authors.affiliation: Department of Pathology andDepartment of Systems Biology, Harvard Medical School, Boston, MA 02115; and.
authors.forename: Anthony
authors.initials: A
authors.lastname: Li
authors.affiliation: Department of Pathology, New York University Medical Center, New York, NY 10016.
authors.forename: Amy
authors.initials: A
authors.lastname: Sun
authors.affiliation: Department of Pathology, New York University Medical Center, New York, NY 10016.
authors.forename: Maryaline
authors.initials: M
authors.lastname: Coffre
authors.affiliation: Department of Pathology, New York University Medical Center, New York, NY 10016.
authors.forename: Sergei B
authors.initials: SB
authors.lastname: Koralov
authors.affiliation: Department of Pathology andDepartment of Systems Biology, Harvard Medical School, Boston, MA 02115; andDivision of Clinical Informatics, Department of Medicine, Beth Israel Deaconess Medical Center, Boston, MA 02215; rarnaout@gmail.com.
authors.forename: Ramy
authors.initials: R
authors.lastname: Arnaout
mesh_terms: D000818:Animals; D001402:B-Lymphocytes / Q000276:immunology*; D022801:Complementarity Determining Regions / Q000235:genetics / Q000276:immunology*; D005260:Female; D059014:High-Throughput Nucleotide Sequencing; D007143:Immunoglobulin Heavy Chains / Q000235:genetics / Q000276:immunology*; D051379:Mice
chemical_list: D022801:Complementarity Determining Regions; D007143:Immunoglobulin Heavy Chains
keywords: development; immunomics; principal component analysis