Mapping the neutralizing epitopes of F18 fimbrial adhesin subunit FedF of enterotoxigenic Escherichia coli (ETEC)

• Published in: Veterinary microbiology 2019-03, Vol.230, p.171-177
• Main Authors: Lu, Ti, Seo, Hyesuk, Moxley, Rodney A., Zhang, Weiping
• Format: Article
• Language: English
• Subjects: Animals; Antibodies; Antibodies, Bacterial - blood; Antibodies, Bacterial - immunology; Antibodies, Neutralizing - blood; Antibodies, Neutralizing - immunology; Antigens, Bacterial - immunology; Bacteria; Bacterial Vaccines - immunology; Cell Line; Colonization; Data processing; Diarrhea; Diarrhea - microbiology; E coli; Enterotoxigenic Escherichia coli - immunology; Enterotoxigenic Escherichia coli - metabolism; Enterotoxins; Epitope Mapping; Escherichia coli; Escherichia coli Infections - immunology; Escherichia coli Infections - prevention & control; Escherichia coli Infections - veterinary; Escherichia coli Proteins - genetics; Escherichia coli Proteins - immunology; ETEC (enterotoxigenic Escherichia coli); F18; FedF; Female; Fimbriae Proteins - genetics; Fimbriae Proteins - immunology; Immunodominance; Immunodominant Epitopes - immunology; Immunodominant Epitopes - isolation & purification; Immunogenicity; Immunoglobulins; Intestine; Mice; Mice, Inbred BALB C; Neutralizing; Neutralizing epitope; PWD (post-weaning diarrhea); Swine; Swine Diseases - microbiology; Vaccine; Vaccine development; Vaccines; Virulence factors; Weaning
• ISSN: 0378-1135; 1873-2542; 1873-2542
• DOI: 10.1016/j.vetmic.2019.02.015

•Neutralizing epitopes are key antigens for vaccines against pig post-weaning diarrhea caused by F18 and K88 fimbrial ETEC.•Immunodominant epitopes of F18 fimbrial adhesin FedF are determined computationally and empirically.•Neutralizing epitopes of F18 FedF adhesin are identified through immunization studies and in vitro assays.•FedF epitopes #3 (IPSSSGTLTCQAGT) and #7 (QPDATGSWYD) are potentially candidates of F18 antigens for PWD vaccine development. K88 and F18 fimbrial enterotoxigenic Escherichia coli (ETEC) are the major causes of post-weaning diarrhea (PWD) in pigs. A vaccine that induces broad immunity to prevent K88 and F18 fimbrial ETEC bacterial attachment and colonization in pig small intestines and to neutralize enterotoxin enterotoxicity would be effective for PWD. Structure-based multiepitope-fusion-antigen (MEFA) technology using a backbone immunogen to present neutralizing epitopes of representing virulence factors capacitates development of broadly protective ETEC vaccines. Neutralizing epitopes have been identified from K88 fimbrial adhesin (FaeG) and enterotoxins but not F18 fimbrial adhesin. In this study, we in silico identified immunodominant epitopes from F18ac fimbrial subunit FedF which plays a critical role in F18 fimbrial adherence, genetically fused each epitope to a carrier, examined immunogenicity of each epitope fusion, and determined epitope-derived antibodies neutralizing activities against F18 fimbrial adherence. Data showed that seven immune-dominant epitopes were identified from FedF subunit. Fused to heterologous human ETEC adhesin subunit CfaB, epitope fusions induced anti-F18 antibodies in subcutaneously immunized mice. Moreover, antibodies derived from each fusion significantly blocked adherence of a F18-fimbrial E. coli bacteria to pig intestinal cell line IPEC-J2. While all seven epitopes exhibited neutralizing activity, results from this study identified FedF epitopes #3 (IPSSSGTLTCQAGT) and #7 (QPDATGSWYD) the most effective for antibodies against F18 fimbrial adherence, and suggested their future application in PWD vaccine development.