Identification and immunogenicity of an immunodominant mimotope of Avibacterium paragallinarum from a phage display peptide library.
Avibacterium paragallinarum is the causative agent of infectious coryza. The protective antigens of this important pathogen have not yet been clearly identified. In this paper, we applied phage display technique to screen the immunodominant mimotopes of a serovar A strain of A. paragallinarum by using a random 12-peptide library, and evaluated the immunogenicity in chickens of the selected mimotope. Polyclonal antibody directed against A. paragallinarum strain 0083 (serovar A) was used as the target antibody and phage clones binding to this target were screened from the 12-mer random peptide library. More than 50% of the phage clones selected in the third round carried the consensus peptide motif sequence A-DP(M)L. The phage clones containing the peptide motif reacted with the target antibody and this interaction could be blocked, in a dose-dependent manner, by A. paragallinarum. One of the peptide sequences, YGLLAVDPLFKP, was selected and the corresponding oligonucleotide sequence was synthesized and then inserted into the expression vector pFliTrx. The recombinant plasmid was transferred into an expression host Escherichia coli GI826 by electroporation, resulting in a recombinant E. coli expressing the peptide on the bacterial surface. Intramuscular injection of the epitope-expressing recombinant bacteria into chickens induced a specific serological response to serovar A. A. paragallinarum. The chickens given the recombinant E. coli showed significant protection against challenge with A. paragallinarum 0083. These results indicated a potential for the use of the mimotope in the development of molecular vaccines for infectious coryza.
Wang
Hongjun
H
Key Laboratory of Preventive Veterinary Medicine of Ministry of Agriculture, College of Veterinary Medicine, China Agricultural University, Beijing 100094, China.
Gao
Yaping
Y
Gong
Yumei
Y
Chen
Xiaoling
X
Liu
Chuan
C
Zhou
Xuemei
X
Blackall
P J
PJ
Zhang
Peijun
P
Yang
Hanchun
H
D000595:Amino Acid Sequence; D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood; D000942:Antigens, Bacterial / Q000276:immunology*; D001435:Bacteriophages; D001483:Base Sequence; D002645:Chickens; D004269:DNA, Bacterial / Q000737:chemistry; D004306:Dose-Response Relationship, Immunologic; D004797:Enzyme-Linked Immunosorbent Assay / Q000662:veterinary; D005260:Female; D006192:Haemophilus Infections / Q000276:immunology / Q000382:microbiology / Q000517:prevention & control / Q000662:veterinary*; D044134:Haemophilus paragallinarum / Q000276:immunology*; D016056:Immunodominant Epitopes; D008969:Molecular Sequence Data; D019151:Peptide Library; D010455:Peptides / Q000276:immunology; D011201:Poultry Diseases / Q000276:immunology / Q000382:microbiology* / Q000517:prevention & control; D012141:Respiratory Tract Infections / Q000276:immunology / Q000382:microbiology / Q000662:veterinary*; D013047:Specific Pathogen-Free Organisms
D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial; D004269:DNA, Bacterial; D016056:Immunodominant Epitopes; D019151:Peptide Library; D010455:Peptides