title: Production and evaluation of chicken egg-yolk-derived antibodies against Campylobacter jejuni colonization-associated proteins.
abstract: Campylobacter jejuni is one of the most important causes of foodborne gastroenteritis. Chickens are considered a reservoir host of C. jejuni, and epidemiological studies have shown that contaminated chicken meat is a primary source of human infection. The objective of this study was to produce chicken egg-yolk-derived antibody (IgY) against the five C. jejuni colonization-associated proteins or CAPs (CadF, FlaA, MOMP, FlpA, and CmeC). Recombinant C. jejuni CAPs were expressed in Escherichia coli and were purified by affinity chromatography. Specific-pathogen-free laying hens were hyperimmunized with each recombinant CAP to induce production of α-CAP-specific IgY. Egg yolks were collected from immunized and nonimmunized hens and were lyophilized to obtain egg-yolk powder (EYP) with or without α-C. jejuni CAP-specific IgY. IgY was purified from EYP, and the antibody response in serum and egg yolk was tested by indirect enzyme-linked immunosorbent assay. The α-C. jejuni CAP-specific IgY levels were significantly (p<0.05) higher in both serum and EYP obtained from immunized hens as compared with the nonimmunized hens. Each α-C. jejuni CAP-specific IgY reacted with the C. jejuni cells and recombinant CAPs as detected by immunofluorescence microscopy and Western blot assays, respectively. We also show that α-CadF, α-MOMP, and α-CmeC IgY significantly reduced adherence of C. jejuni to the chicken hepatocellular carcinoma (LMH) cells, suggesting that these α-C. jejuni CAP-specific IgY may be useful as a passive immunotherapeutic to reduce C. jejuni colonization in chickens.
authors.affiliation: Department of Veterinary Microbiology and Pathology, Washington State University, Pullman, Washington 99164, USA.
authors.forename: Salma R
authors.initials: SR
authors.lastname: Al-Adwani
authors.forename: Rocio
authors.initials: R
authors.lastname: Crespo
authors.forename: Devendra H
authors.initials: DH
authors.lastname: Shah
mesh_terms: D000818:Animals; D000907:Antibodies, Bacterial / Q000096:biosynthesis / Q000097:blood; D000918:Antibody Specificity; D000942:Antigens, Bacterial / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification / Q000378:metabolism; D001426:Bacterial Proteins / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism; D002169:Campylobacter Infections / Q000276:immunology / Q000382:microbiology / Q000662:veterinary; D016123:Campylobacter jejuni / Q000276:immunology*; D045744:Cell Line, Tumor; D002645:Chickens; D004530:Egg Yolk / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay / Q000662:veterinary; D005260:Female; D006801:Humans; D007114:Immunization / Q000662:veterinary; D007136:Immunoglobulins / Q000097:blood / Q000276:immunology*; D008856:Microscopy, Fluorescence / Q000662:veterinary; D011994:Recombinant Proteins; D013047:Specific Pathogen-Free Organisms
chemical_list: D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial; D001426:Bacterial Proteins; C016512:IgY; D007136:Immunoglobulins; D011994:Recombinant Proteins