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ARS Home » Southeast Area » Athens, Georgia » U.S. National Poultry Research Center » Poultry Microbiological Safety & Processing Research » Research » Publications at this Location » Publication #321570

Research Project: Pre-Harvest Interventions For Application During Poultry Production To Reduce Food-Borne Bacterial Pathogens

Location: Poultry Microbiological Safety & Processing Research

Title: Epitope mapping of campylobacter jejuni flagellar capping protein (FliD) by chicken (gallus gallus domesticus) sera

Author
item Yeh, Hung-yueh
item Telli, Arife Ezgi - Selcuk University
item Jagne, Jarra - Cornell University - New York
item Benson, Christopher - University Of Georgia
item Hiett, Kelli
item Line, John - Eric

Submitted to: Comparative Immunology Microbiology and Infectious Diseases
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 10/18/2016
Publication Date: 10/18/2016
Citation: Yeh, H., Telli, A., Jagne, J.G., Benson, C.L., Hiett, K.L., Line, J.E. 2016. Epitope mapping of campylobacter jejuni flagellar capping protein (FliD) by chicken (gallus gallus domesticus) sera. Comparative Immunology Microbiology and Infectious Diseases. 49:76-81.

Interpretive Summary: Campylobacter jejuni is an important foodborne pathogen, which causes human acute bacterial gastroenteritis worldwide. Poultry products are considered as the major source of this bacterium for human infection. To prevent human infection, vaccination of chickens is one of powerful means to reduce this bacterium in broiler chicken gut and minimize contamination in poultry. The flagellar capping protein (FliD) has been implicated for microorganism colonization in the host gastrointestinal tract as well as inducing protective antibodies. 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 commercial field chickens. The results 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 strongly reacted to more than three chicken sera 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.

Technical 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 strongly reacted to more than three chicken sera 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.