Vaccine efficacy of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) from Edwardsiella ictaluri against E. tarda in tilapia.
Glyceraldehyde-3-phosphate dehydrogenase (GAPDH), derived from the outer-membrane protein (OMP) fraction, has been used as a potential candidate for vaccine development. The gene-encoding 37 kDa GAPDH outer membrane protein (OMP) from Edwardsiella ictaluri was amplified using polymerase chain reaction (PCR) and was cloned and expressed in Escherichia coli BL21 (DE3). Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), Western blotting, and nucleotide and amino acid sequencing were used to analyze the expressed antigenic protein and gene encoding this protein. Comparative DNA and protein sequence analysis of GAPDH from E. ictaluri GAPDHs from several Gram-negative bacterial species within the Enterobacteriaceae family revealed that the GAPDHs within this group are highly conserved and share a sequence similarity of 75-100% with E. ictaluri GDPDH. Rabbit antiserum raised against the E. ictaluri recombinant GAPDH (rGAPDH) protein recognized purified GADPH, indicating that it has a strong immunogenicity. Tilapia fish were intraperitoneally immunized with formalin-killed E. ictaluri whole cells, and rGAPDH (30 μg fish(-1)) from E. ictaluri, both of which were emulsified in ISA 763A adjuvant. At 3 months after immunization, fish were challenged with the E. tarda strain to assess vaccine efficacy; the relative percent survival (RPS) values were found to exceed 71.4%. The specific mean antibody titer log2 level of groups vaccinated with rGAPDH at 3 months was significantly higher than that of non-vaccinated fish (control group). Therefore, this recombinant protein can be considered a multi-purpose candidate vaccine against several pathogenic bacteria.
Department of Tropical Agriculture and International Cooperation.
Thanh
T
Trung Cao
Ming-An
MA
Tsai
Chung-Da
CD
Yang
Pei-Chyi
PC
Wang
Tsun-Yung
TY
Kuo
Hsu-Chung
HC
Gabriel Chen
Shih-Chu
SC
Chen
D000276:Adjuvants, Immunologic / Q000008:administration & dosage; D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood; D001428:Bacterial Vaccines / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology*; D003001:Cloning, Molecular; D020610:Edwardsiella ictaluri / Q000201:enzymology*; D020609:Edwardsiella tarda / Q000276:immunology*; D004756:Enterobacteriaceae Infections / Q000276:immunology / Q000517:prevention & control*; D004926:Escherichia coli / Q000235:genetics; D005393:Fish Diseases / Q000276:immunology / Q000517:prevention & control*; D005986:Glyceraldehyde 3-Phosphate / Q000276:immunology*; D016133:Polymerase Chain Reaction; D017386:Sequence Homology, Amino Acid; D016019:Survival Analysis; D017210:Tilapia; D014614:Vaccines, Synthetic / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology
D000276:Adjuvants, Immunologic; D000907:Antibodies, Bacterial; D001428:Bacterial Vaccines; D014614:Vaccines, Synthetic; D005986:Glyceraldehyde 3-Phosphate