Functional pentameric formation via coexpression of the Escherichia coli heat-labile enterotoxin B subunit and its fusion protein subunit with a neutralizing epitope of ApxIIA exotoxin improves the mucosal immunogenicity and protection against challenge by Actinobacillus pleuropneumoniae.
A coexpression strategy in Saccharomyces cerevisiae using episomal and integrative vectors for the Escherichia coli heat-labile enterotoxin B subunit (LTB) and a fusion protein of an ApxIIA toxin epitope produced by Actinobacillus pleuropneumoniae coupled to LTB, respectively, was adapted for the hetero-oligomerization of LTB and the LTB fusion construct. Enzyme-linked immunosorbent assay (ELISA) with GM1 ganglioside indicated that the LTB fusion construct, along with LTB, was oligomerized to make the functional heteropentameric form, which can bind to receptors on the mucosal epithelium. The antigen-specific antibody titer of mice orally administered antigen was increased when using recombinant yeast coexpressing the pentameric form instead of recombinant yeast expressing either the LTB fusion form or antigen alone. Better protection against challenge infection with A. pleuropneumoniae was also observed for coexpression in recombinant yeast compared with others. The present study clearly indicated that the coexpression strategy enabled the LTB fusion construct to participate in the pentameric formation, resulting in an improved induction of systemic and mucosal immune responses.
Institute for Molecular Biology and Genetics, Center for Fungal Pathogenesis, Chonbuk National University, Dukjindong 664-14, Jeonju, Chonbuk 561-756, South Korea.
Jung-Mi
JM
Kim
Seung-Moon
SM
Park
Jung-Ae
JA
Kim
Jin-Ah
JA
Park
Min-Hee
MH
Yi
Nan-Sun
NS
Kim
Jong-Lye
JL
Bae
Sung Goo
SG
Park
Yong-Suk
YS
Jang
Moon-Sik
MS
Yang
Dae-Hyuk
DH
Kim
D000189:Actinobacillus Infections / Q000276:immunology / Q000517:prevention & control*; D016977:Actinobacillus pleuropneumoniae / Q000235:genetics / Q000276:immunology*; D058956:Administration, Mucosal; D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood; D001426:Bacterial Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D001427:Bacterial Toxins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D004195:Disease Models, Animal; D004768:Enterotoxins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D000939:Epitopes / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics; D029968:Escherichia coli Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D022361:Escherichia coli Vaccines / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology*; D006460:Hemolysin Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D051379:Mice; D055503:Protein Multimerization; D021122:Protein Subunits / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics; D016019:Survival Analysis
D000907:Antibodies, Bacterial; C085361:ApxII toxin, bacteria; D001426:Bacterial Proteins; D001427:Bacterial Toxins; D004768:Enterotoxins; D000939:Epitopes; D029968:Escherichia coli Proteins; D022361:Escherichia coli Vaccines; D006460:Hemolysin Proteins; D021122:Protein Subunits; D011993:Recombinant Fusion Proteins; C027538:heat-labile enterotoxin, E coli