posted by Maria Lucrecia Alvarez at
1:50 PM
Monday, October 08, 2007
Saturday, September 15, 2007
SUBUNIT VACCINE DEVELOPED IN TOMATO TARGETED AGAINST PLAGUE AND ITS IMMUNOGENICITY IN MICE
1 ) THE LAB
DR. CARDINEAU'S LAB
Center for Infectious Diseases and Vaccinology (CIDV). The Biodesign Institute at Arizona State University. 1001 S. McAllister Ave., Tempe, AZ 85287, USA.
____________________________________________________
DIRECTOR:
Dr. Guy Cardineau
e-mail: guy.cardineau@asu.edu
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POSTDOCTORAL RESEARCH ASSOCIATE:
Dr. Maria Lucrecia Alvarez
e-mail: lucrecia.alvarez@asu.edu
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RESEARCH TECHNICIANS:
Heidi Pinyerd
Federico Martin
UNDERGRADUATED STUDENTS:
Iason Crisante
Angela Rojas
Sachi Dale
Michael Ewing
2 ) RESULTS OBTAINED
ABSTRACT
Yersinia pestis, the causative agent of plague, is one of the most virulent bacteria ever known but presently there are no approved vaccines for protection against it. Our goal was to obtain a vaccine that would address several issues: ease of delivery, mucosal efficacy, safety, rapid scalability, and cost. We developed a novel production and delivery system for a plague vaccine consisting of a Y. pestis F1-V antigen fusion protein expressed in tomato and orally delivered as a powder. We selected elite tomato plants expressing high levels of the F1-V fusion protein in fruit. The immunogenicity of these F1-V transgenic tomatoes was confirmed in BALB/c mice that were primed with subcutaneous bacterially produced F1-V and boosted orally with transgenic tomatoes. The humoral response was dominated by IgG1 over IgG2a, the preferred response to confer protection against plague.
INTRODUCTION
Plague or “Black Death” is an infection of rodents caused by the bacterium Yersinia pestis and accidentally transmitted to humans by the bite of infected fleas. The primary vector is Xenopsylla cheopis, the rodent flea with a predilection for rats, that bites humans after the death of the infected rodents. The disease follows urban and rural cycles in nature but Yersinia pestis could also be aerosolized and used as a bioterrorist agent.
The History of Bioterrorism: Plague
Video Source: http://www.cdc.gov/
GOAL
Develop a plant-derived oral vaccine in tomato for the prevention of bubonic and pneumonic plague.
RESULTS
A) EXPRESSION OF F1-V FUSION PROTEIN IN TOMATO LEAVES AND FRUIT
B) MOLECULAR ANALYSIS
CONCLUSIONS
1) Transgenic tomato plants expressing high levels of the fusion protein F1-V in fruits (3 to 8% of total soluble protein, 600 to 1600 µg /g freeze-dried tomato) were obtained as a possible plague vaccine.
2) Most of the second generation elite line plants have a low gene copy number (2-5) (Southern-blot) and they showed only one RNA transcript of the expected size in Northen-blot analysis.
3) The immunogenicity of the orally administrated F1-V transgenic tomatoes was confirmed by prime-boost experiments in mice. The humoral response was dominated by IgG1 over IgG2a. This suggests a type 2 T-Helper immune response mediated by antibodies that is the optimum response to confer protection against plague.
4) Preliminary results: mice primed with s.c. bacterially derived F1-V and boosted with transgenic tomato were protected against a challenge with s.c. Y. pestis, which was done almost 18 months after the last boosting.
posted by Maria Lucrecia Alvarez at
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Wednesday, June 14, 2006
Recent Publications
- Alvarez ML, Pinyerd HL, Crisantes JD, Rigano MM, Pinkhasov J, Walmsley AM, Mason HS, Cardineau GA (2006). Plant-made subunit vaccine against pneumonic and bubonic plague is orally immunogenic in mice. Vaccine 24(14): 2477-90. To view this article, click here .
- Alvarez ML, Pinyerd HL, Topal E, Cardineau GA (2008). P19-dependent and P19-independent reversion of f1-v gene silencing in tomato. Plant mol Biol (in press).
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