title: Epitope mapping of Campylobacter jejuni flagellar capping protein (FliD) by chicken (Gallus gallus domesticus) sera.
abstract: Campylobacter jejuni, a Gram-negative rod, is a zoonotic pathogen associated with human acute bacterial gastroenteritis worldwide. The flagellum, composed of more than 35 proteins, is responsible for colonization of C. jejuni in the host gastrointestinal tract as well as inducing protective antibodies against the homologous serotype. In our previous study, we demonstrated that the flagellar capping protein (FliD) is an immunodominant protein that reacted strongly to sera from field chickens. In this communication, we mapped linear immunoreactive epitopes on FliD using a set of 158 synthetic peptides of 15-mer overlapping with 11 amino acid residues on peptide microarrays with sera from field chickens. The results from peptide microarrays showed (1) no cross-reactivity of the immobilized peptides with the secondary anti-chicken antibody in the control incubation, and (2) heterogeneous patterns of sera reacting to the immobilized peptides. The peptides that reacted to more than three chicken sera and had higher averages of fluorescence units were selected for further validation by the peptide ELISA. The results showed peptides 24, 91 and 92 had relatively high reactivity and less variation among 64 individual serum samples, indicating these peptides represented the shared immunodominant epitopes on the C. jejuni FliD protein. These peptides were also recognized by sera from chickens immunized with the purified recombinant FliD protein. The findings of the specific shared linear immunodominant epitopes on FliD in this study provide a rationale for further evaluation to determine their utility as epitope vaccines covering multiple serotypes for chicken immunization, and subsequently, for providing safer poultry products for human consumption.
authors.affiliation: United States Department of Agriculture, Agricultural Research Service, U.S. National Poultry Research Center, Poultry Microbiological Safety and Processing Research Unit, 950 College Station Road, Athens, GA 30605-2720, USA. Electronic address: hungyueh.yeh@ars.usda.gov.
authors.forename: Hung-Yueh
authors.initials: HY
authors.lastname: Yeh
authors.affiliation: Laboratory of Food Hygiene and Technology, Faculty of Veterinary Medicine, University of Selcuk, 42003, Konya, Turkey.
authors.forename: Arife Ezgi
authors.initials: AE
authors.lastname: Telli
authors.affiliation: Animal Health Diagnostic Center, Cornell University, Ithaca, NY 14853, USA.
authors.forename: Jarra F
authors.initials: JF
authors.lastname: Jagne
authors.affiliation: University of Georgia, Athens, GA 30605, USA.
authors.forename: Christopher L
authors.initials: CL
authors.lastname: Benson
authors.affiliation: United States Department of Agriculture, Agricultural Research Service, U.S. National Poultry Research Center, Poultry Microbiological Safety and Processing Research Unit, 950 College Station Road, Athens, GA 30605-2720, USA.
authors.forename: Kelli L
authors.initials: KL
authors.lastname: Hiett
authors.affiliation: United States Department of Agriculture, Agricultural Research Service, U.S. National Poultry Research Center, Poultry Microbiological Safety and Processing Research Unit, 950 College Station Road, Athens, GA 30605-2720, USA.
authors.forename: John E
authors.initials: JE
authors.lastname: Line
mesh_terms: D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood / Q000276:immunology; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D016123:Campylobacter jejuni / Q000276:immunology*; D002645:Chickens / Q000276:immunology; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay; D018604:Epitope Mapping; D005407:Flagella / Q000737:chemistry; D006801:Humans; D007114:Immunization; D016056:Immunodominant Epitopes; D010446:Peptide Fragments / Q000737:chemistry / Q000276:immunology; D040081:Protein Array Analysis; D011994:Recombinant Proteins / Q000276:immunology
chemical_list: D000907:Antibodies, Bacterial; D001426:Bacterial Proteins; D016056:Immunodominant Epitopes; D010446:Peptide Fragments; D011994:Recombinant Proteins; C076733:FlaD protein, Bacteria
keywords: Campylobacter jejuni; Epitope mapping; Flagellar capping protein; FliD; Foodborne pathogen; Zoonosis