Pentavalent single-domain antibodies reduce Campylobacter jejuni motility and colonization in chickens

PLoS One. 2013 Dec 31;8(12):e83928. doi: 10.1371/journal.pone.0083928. eCollection 2013.

Abstract

Campylobacter jejuni is the leading cause of bacterial foodborne illness in the world, with symptoms ranging from acute diarrhea to severe neurological disorders. Contaminated poultry meat is a major source of C. jejuni infection, and therefore, strategies to reduce this organism in poultry, are expected to reduce the incidence of Campylobacter-associated diseases. We have investigated whether oral administration of C. jejuni-specific single-domain antibodies would reduce bacterial colonization levels in chickens. Llama single-domain antibodies specific for C. jejuni were isolated from a phage display library generated from the heavy chain IgG variable domain repertoire of a llama immunized with C. jejuni flagella. Two flagella-specific single-domain antibodies were pentamerized to yield high avidity antibodies capable of multivalent binding to the target antigen. When administered orally to C. jejuni-infected two-day old chicks, the pentabodies significantly reduced C. jejuni colonization in the ceca. In vitro, the motility of the bacteria was also reduced in the presence of the flagella-specific pentabodies, suggesting the mechanism of action is through either direct interference with flagellar motility or antibody-mediated aggregation. Fluorescent microscopy and Western blot analyses revealed specific binding of the anti-flagella pentabodies to the C. jejuni flagellin.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Administration, Oral
  • Amino Acid Sequence
  • Animals
  • Antibodies, Bacterial / administration & dosage
  • Antibodies, Bacterial / immunology
  • Antibodies, Bacterial / pharmacology*
  • Blotting, Western
  • Camelids, New World
  • Campylobacter Infections / immunology
  • Campylobacter Infections / microbiology
  • Campylobacter Infections / prevention & control
  • Campylobacter Infections / veterinary*
  • Campylobacter jejuni / immunology*
  • Cecum / microbiology
  • Cell Movement / physiology*
  • Cell Surface Display Techniques
  • Chickens
  • Enzyme-Linked Immunosorbent Assay
  • Female
  • Flagella / immunology
  • Flagella / metabolism
  • Flagellin / immunology
  • Flagellin / metabolism
  • Gastrointestinal Tract / immunology
  • Gastrointestinal Tract / microbiology*
  • Immunization
  • Male
  • Microscopy, Fluorescence
  • Molecular Sequence Data
  • Poultry Diseases / immunology
  • Poultry Diseases / microbiology
  • Poultry Diseases / prevention & control*
  • Sequence Homology, Amino Acid
  • Single-Domain Antibodies / administration & dosage
  • Single-Domain Antibodies / immunology
  • Single-Domain Antibodies / pharmacology*
  • Surface Plasmon Resonance

Substances

  • Antibodies, Bacterial
  • Single-Domain Antibodies
  • Flagellin

Grant support

The authors are grateful to Dow AgroSciences and AbCelex Technologies for partially funding this project. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.