Identifying functional anti-Staphylococcus aureus antibodies by sequencing antibody repertoires of patient plasmablasts

Clin Immunol. May-Jun 2014;152(1-2):77-89. doi: 10.1016/j.clim.2014.02.010. Epub 2014 Mar 1.

Abstract

Infection by Staphylococcus aureus is on the rise, and there is a need for a better understanding of host immune responses that combat S. aureus. Here we use DNA barcoding to enable deep sequencing of the paired heavy- and light-chain immunoglobulin genes expressed by individual plasmablasts derived from S. aureus-infected humans. Bioinformatic analysis of the antibody repertoires revealed clonal families of heavy-chain sequences and enabled rational selection of antibodies for recombinant expression. Of the ten recombinant antibodies produced, seven bound to S. aureus, of which four promoted opsonophagocytosis of S. aureus. Five of the antibodies bound to known S. aureus cell-surface antigens, including fibronectin-binding protein A. Fibronectin-binding protein A-specific antibodies were isolated from two independent S. aureus-infected patients and mediated neutrophil killing of S. aureus in in vitro assays. Thus, our DNA barcoding approach enabled efficient identification of antibodies involved in protective host antibody responses against S. aureus.

Keywords: Antibody; Infection; Staphylococcus aureus.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • 3T3 Cells
  • Adhesins, Bacterial / immunology
  • Animals
  • Antibodies, Bacterial / immunology*
  • Antibody Formation / immunology
  • Base Sequence
  • DNA Barcoding, Taxonomic
  • Humans
  • Immunoglobulin Heavy Chains / genetics*
  • Immunoglobulin Heavy Chains / immunology
  • Immunoglobulin Light Chains / genetics*
  • Immunoglobulin Light Chains / immunology
  • Mice
  • Neutrophils / immunology
  • Phagocytosis / immunology
  • Recombinant Proteins / immunology
  • Sequence Analysis, DNA / methods
  • Staphylococcal Infections / immunology*
  • Staphylococcus aureus / immunology*

Substances

  • Adhesins, Bacterial
  • Antibodies, Bacterial
  • Immunoglobulin Heavy Chains
  • Immunoglobulin Light Chains
  • Recombinant Proteins
  • fibronectin-binding proteins, bacterial