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

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

Author information

  • 1Division of Immunology and Rheumatology, Stanford University, CCSR 4135, 269 Campus Dr., Stanford, CA 94305, USA; VA Palo Alto Health Care System, 3801 Miranda Ave, Palo Alto, CA 94304, USA; Stanford Immunology Program, Stanford University School of Medicine, Stanford, CA 94305, USA.
  • 2Division of Immunology and Rheumatology, Stanford University, CCSR 4135, 269 Campus Dr., Stanford, CA 94305, USA; VA Palo Alto Health Care System, 3801 Miranda Ave, Palo Alto, CA 94304, USA.
  • 3Division of Immunology and Rheumatology, Stanford University, CCSR 4135, 269 Campus Dr., Stanford, CA 94305, USA.
  • 4Division of Immunology and Rheumatology, Stanford University, CCSR 4135, 269 Campus Dr., Stanford, CA 94305, USA; VA Palo Alto Health Care System, 3801 Miranda Ave, Palo Alto, CA 94304, USA; Stanford Immunology Program, Stanford University School of Medicine, Stanford, CA 94305, USA. Electronic address: wrobins@stanford.edu.

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.

Published by Elsevier Inc.

KEYWORDS:

Antibody; Infection; Staphylococcus aureus

PMID:
24589749
[PubMed - indexed for MEDLINE]
PMCID:
PMC4066023
Free PMC Article
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