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DEVELOPMENT OF AN EDIBLE RABIES VACCINE IN MAIZE, USING VNUKOVO STRAIN

DEVELOPMENT OF AN EDIBLE RABIES VACCINE IN MAIZE, USING VNUKOVO STRAIN. Elizabeth Loza Rubio, NCVM-INIFAP, Mexico Edith Rojas Anaya NCVM-INIFAP Luis Gómez Nuñez NCVM-INIFAP Teresa Olivera Flores, UNAM Miguel Gómez-Lim, CINVESTAV.

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DEVELOPMENT OF AN EDIBLE RABIES VACCINE IN MAIZE, USING VNUKOVO STRAIN

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  1. DEVELOPMENT OF AN EDIBLE RABIES VACCINE IN MAIZE, USING VNUKOVO STRAIN Elizabeth Loza Rubio, NCVM-INIFAP, Mexico Edith Rojas Anaya NCVM-INIFAP Luis Gómez Nuñez NCVM-INIFAP Teresa Olivera Flores, UNAM Miguel Gómez-Lim, CINVESTAV

  2. Plants are natural bioreactors and are effective for production of recombinant proteins and antigens. There are already several plant-produced proteins on the market, including one at large scale. Proof of concept has been well established for the production of a wide range of many therapeutic proteins, including vaccines for humans and animals.

  3. POTENTIAL ADVANTAGES FOR THE PRODUCTION OF RECOMBINANT PROTEINS IN PLANTS Plant systems are more economical than industrial facilities using fermentation or bioreactor systems. The technology is already available for harvesting and processing plants and products on a large scale. Plants can be directed to target proteins into intracellular compartments in which they are more stable. Health risks arising from contamination with potential human pathogens or toxins are minimized

  4. SOME ADVANTAGES OF ANTIGENS EXPRESSED IN TRANSGENIC PLANTS Oral delivery of vaccines is an attractive alternative to injection, largely for reasons of low cost and easy administration. The sowing process, harvests, storage and transport is such as for the traditional plants They are more stable to the hot weather It does not require the culture of the infectious agent Multivalent vaccines can be developed

  5. Several cereals, and in particular the maize have been the system of choice to express antigenic proteins, which can be stored for long periods without excessive deterioration It constitutes a major proportion of animal diet, and heat and pressure treatments are not necessary

  6. OBJECTIVE The goal of this work was to obtain transgenic maize expressing the rabies virus G protein of Vnukovo strain and to evaluate immunogenicity in mice, by oral route.

  7. promCMV RV-gp PoliA Amp ORI pWR-21 (5610 PB) Fodor I et al, Acta Vet Hung 2000, 48:229-326

  8. MAR promUbi Gen G Ter pUCpSS MAR Vector pGHNC5 Gen N Ter CaMV 35S CaMV pUCpSSrabN 4.9 Kb CONSTRUCTION OF VECTORS

  9. DETECTION OF G GENE OF RABIES VIRUS IN DIFFERENT LINES OF TRANSFORMED MAIZE, BY PCR Lane 1, MW; lane 2 Rabies virus; lane 3, Non transformed maize; Lane 4, 5, 6, transformed maize

  10. DETECTION OF PROTEIN, USING WESTERN BLOT 1 2 3 4 5 6 7 69 kDa 65 kDa Lane 1-4, G protein expressed by transformed maize; lane 5, non transformed maize; lane 6, purified G protein; lane 7, MW. 1 2 3 4 5 6 7 1 2 3 4 5 6 7 69 KDa 69 KDa Figure 4. Detection of G protein, using WB. Lane 1-4, G protein expressed by transformed maize;lane 5, non transformed maize;purified G protein; lane 7, MW. Figure 4. Detection of G protein, using WB. Lane 1-4, G protein expressed by transformed maize;lane 5, non transformed maize;purified G protein; lane 7, MW.

  11. Virus neutralizing anti-rabies antibodies in mice. Group 1. Mice were immunized with a modified live rabies virus vaccine; Group 2. Adult mice were feed with 50 mg of G protein expressed in maize (orally); Group 3. Control non-transformed corn.

  12. Survivorship of mice Survivorship in mice immunized with three different treatments and challenged intracraneally with 100 LD50 of a vampire-bat strain.

  13. CONCLUSION It was possible to transform corn using biolistics The presence of G gene and its product was detected in transformed plants Co-integration percentage in analysed plants was of 93.3%. The amount of G protein detected in grains was approximately an average of 1% G protein of Vnukovo strain, expressed in transgenic maize may work as oral immunogen against an heterologous challenge CONCLUSIONS The edible vaccine was able to induce neutralizing antibodies in both mono and polygastric models (Fig. 5 and 7), which were able to protect mice (100%) and ovines (>80%) against challenge with a vampire bat-strain, which is one of the main reservoirs in Latin America. G protein expressed in transgenic maize may work as oral immunogen against rabies It is the first time that this kind of vaccines is evaluated and could work in a ruminant species. ACKNOWLEDGMENT PROJECT G34635B, CONACYT MEXICO National Foundation for Infectious Disease for the Travel Grant

  14. Merçi bien! Thank you! Спасибо Grazie 謝謝 ¡Gracias! Danke!

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