Surface-displayed expression of a neutralizing epitope of ApxIIA exotoxin in Saccharomyces cerevisiae and oral administration of it for protective immune responses against challenge by Actinobacillus pleuropneumoniae.
A neutralizing epitope fragment of ApxIIA toxin (ApxIIA#5) of the Korean Actinobacillus pleuropneumoniae serotype 2 strain was expressed and immobilized on the cell surface of Saccharomyces cerevisiae for efficient vaccine development. Expression of ApxIIA#5 was confirmed by Western blot analysis using cell-wall proteins, and the surface display of ApxIIA#5 was further visualized under confocal microscopy. Quantitative ELISA revealed that the recombinant ApxIIA#5 directed to the cell surface consisted of approximately 16% cell-wall proteins, estimated to be 35 mg of ApxIIA#5 protein per liter of cultured cells. An immunoassay revealed that antigen-specific antibodies against ApxIIA#5 were present in the sera of mice fed recombinant ApxIIA#5-expressing yeast, but not in mice fed the wild-type nor the vector-only transformant. Moreover, the mice fed the recombinant epitope-expressing yeast were protected from injection of a lethal dose of A. pleuropneumoniae.
Kim
Jung-Mi
JM
Institute for Molecular Biology and Genetics, Center for Fungal Pathogenesis, Chonbuk National University, Korea.
Jung
Dea-Im
DI
Eom
Yoo Jeong
YJ
Park
Seung-Moon
SM
Yoo
Han-Sang
HS
Jang
Yong-Suk
YS
Yang
Moon-Sik
MS
Kim
Dae-Hyuk
DH
D016977:Actinobacillus pleuropneumoniae / Q000276:immunology*; D000284:Administration, Oral; D000818:Animals; D000918:Antibody Specificity; D001426:Bacterial Proteins / Q000276:immunology*; D015153:Blotting, Western; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000008:administration & dosage / Q000235:genetics* / Q000276:immunology*; D005260:Female; D015870:Gene Expression; D006460:Hemolysin Proteins / Q000276:immunology*; D051379:Mice; D008853:Microscopy; D010957:Plasmids / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D014170:Transformation, Genetic; D014611:Vaccination
C085361:ApxII toxin, bacteria; D001426:Bacterial Proteins; D000939:Epitopes; D006460:Hemolysin Proteins