title: Pentavalent single-domain antibodies reduce Campylobacter jejuni motility and colonization in chickens.
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.
authors.affiliation: AbCelex Technologies Inc., Toronto, Ontario, Canada.
authors.forename: Ali
authors.initials: A
authors.lastname: Riazi
authors.affiliation: Human Health Therapeutics, National Research Council of Canada, Ottawa, Ontario, Canada.
authors.forename: Philippa C R
authors.initials: PC
authors.lastname: Strong
authors.affiliation: Human Health Therapeutics, National Research Council of Canada, Ottawa, Ontario, Canada.
authors.forename: Russell
authors.initials: R
authors.lastname: Coleman
authors.affiliation: Human Health Therapeutics, National Research Council of Canada, Ottawa, Ontario, Canada.
authors.forename: Wangxue
authors.initials: W
authors.lastname: Chen
authors.affiliation: Human Health Therapeutics, National Research Council of Canada, Ottawa, Ontario, Canada.
authors.forename: Tomoko
authors.initials: T
authors.lastname: Hirama
authors.affiliation: Human Health Therapeutics, National Research Council of Canada, Ottawa, Ontario, Canada.
authors.forename: Henk
authors.initials: H
authors.lastname: van Faassen
authors.affiliation: Dow AgroSciences, Indianapolis, Indiana, United States of America.
authors.forename: Matthew
authors.initials: M
authors.lastname: Henry
authors.affiliation: Human Health Therapeutics, National Research Council of Canada, Ottawa, Ontario, Canada.
authors.forename: Susan M
authors.initials: SM
authors.lastname: Logan
authors.affiliation: Centennial Centre for Interdisciplinary Science, Department of Biological Sciences and Alberta Glycomics Centre, The University of Alberta, Edmonton, Alberta, Canada.
authors.forename: Christine M
authors.initials: CM
authors.lastname: Szymanski
authors.affiliation: Human Health Therapeutics, National Research Council of Canada, Ottawa, Ontario, Canada ; Centennial Centre for Interdisciplinary Science, Department of Biological Sciences and Alberta Glycomics Centre, The University of Alberta, Edmonton, Alberta, Canada.
authors.forename: Roger
authors.initials: R
authors.lastname: Mackenzie
authors.affiliation: Human Health Therapeutics, National Research Council of Canada, Ottawa, Ontario, Canada ; School of Environmental Sciences, University of Guelph, Guelph, Ontario, Canada ; Department of Biology, Carleton University, Ottawa, Ontario, Canada.
authors.forename: Mehdi Arbabi
authors.initials: MA
authors.lastname: Ghahroudi
mesh_terms: D000284:Administration, Oral; D000595:Amino Acid Sequence; D000818:Animals; D000907:Antibodies, Bacterial / Q000008:administration & dosage / Q000276:immunology / Q000494:pharmacology*; D015153:Blotting, Western; D002161:Camelids, New World; D002169:Campylobacter Infections / Q000276:immunology / Q000382:microbiology / Q000517:prevention & control / Q000662:veterinary*; D016123:Campylobacter jejuni / Q000276:immunology*; D002432:Cecum / Q000382:microbiology; D002465:Cell Movement / Q000502:physiology*; D061505:Cell Surface Display Techniques; D002645:Chickens; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D005407:Flagella / Q000276:immunology / Q000378:metabolism; D005408:Flagellin / Q000276:immunology / Q000378:metabolism; D041981:Gastrointestinal Tract / Q000276:immunology / Q000382:microbiology*; D007114:Immunization; D008297:Male; D008856:Microscopy, Fluorescence; D008969:Molecular Sequence Data; D011201:Poultry Diseases / Q000276:immunology / Q000382:microbiology / Q000517:prevention & control*; D017386:Sequence Homology, Amino Acid; D061905:Single-Domain Antibodies / Q000008:administration & dosage / Q000276:immunology / Q000494:pharmacology*; D020349:Surface Plasmon Resonance
chemical_list: D000907:Antibodies, Bacterial; D061905:Single-Domain Antibodies; D005408:Flagellin