Poliovirus replicons encoding the B subunit of Helicobacter pylori urease elicit a Th1 associated immune response.
The development of a vaccine for Helicobacter pylori is a key strategy for reducing the worldwide prevalence of H. pylori infection. Although immunization with recombinant B subunit of H. pylori urease (ureB) has yielded promising results, for the most part, these studies relied on the use of strong adjuvant, cholera toxin, precluding the use in humans. Thus, the development of new vaccine strategies for H. pylori is essential. Previous studies from our laboratory have described a vaccine vector based on poliovirus in which foreign genes are substituted for the poliovirus capsid genes. The genomes encoding foreign proteins (replicons) are encapsidated into authentic poliovirions by providing the capsids in trans. To test the utility of replicons as a vaccine vector for H. pylori, a replicon was constructed which encodes ureB. Expression of ureB in cells from the replicon was demonstrated by metabolic labeling followed by immunoprecipitation with anti-urease antibodies. To investigate the immunogenicity of the replicons, mice containing the transgene for the receptor for poliovirus were immunized via the intramuscular route. Mice given three doses of replicons did not develop substantial antibodies to ureB as determined by Western blot analysis using lysates from H. pylori. In contrast, mice given two doses of replicon followed by a single injection of recombinant ureB developed serum antibodies to ureB which were predominately IgG2a. Splenic lymphocytes from mice immunized with replicons alone, or replicons plus recombinant ureB produced abundant interferon-gamma and no detectable interleukin-4 upon stimulation with recombinant ureB. These results establish that poliovirus replicons encoding H. pylori ureB are immunogenic and induce primarily a T helper 1 associated immune response.
Novak
M J
MJ
Department of Microbiology, University of Alabama at Birmingham 35294, USA.
Smythies
L E
LE
McPherson
S A
SA
Smith
P D
PD
Morrow
C D
CD
D000818:Animals; D000907:Antibodies, Bacterial / Q000096:biosynthesis; D001426:Bacterial Proteins / Q000096:biosynthesis / Q000235:genetics / Q000627:therapeutic use; D001428:Bacterial Vaccines / Q000235:genetics / Q000276:immunology; D015153:Blotting, Western; D002213:Capsid / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D036022:Capsid Proteins; D004269:DNA, Bacterial / Q000235:genetics / Q000627:therapeutic use; D004279:DNA, Viral / Q000276:immunology; D016481:Helicobacter Infections / Q000276:immunology* / Q000517:prevention & control*; D016480:Helicobacter pylori / Q000235:genetics* / Q000276:immunology*; D007371:Interferon-gamma / Q000378:metabolism; D015847:Interleukin-4 / Q000378:metabolism; D051379:Mice; D008822:Mice, Transgenic; D010452:Peptide Biosynthesis; D010455:Peptides / Q000235:genetics / Q000276:immunology; D017955:Poliovirus / Q000235:genetics* / Q000276:immunology; D011233:Precipitin Tests; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000097:blood / Q000235:genetics / Q000276:immunology; D012093:Replicon / Q000276:immunology*; D018417:Th1 Cells / Q000276:immunology*; D014510:Urease / Q000235:genetics* / Q000276:immunology*; D014611:Vaccination / Q000379:methods
D000907:Antibodies, Bacterial; D001426:Bacterial Proteins; D001428:Bacterial Vaccines; D036022:Capsid Proteins; D004269:DNA, Bacterial; D004279:DNA, Viral; D010455:Peptides; D011993:Recombinant Fusion Proteins; C060339:VP4 protein, Rotavirus; D015847:Interleukin-4; D007371:Interferon-gamma; D014510:Urease