DEVELOPMENT OF AN EDIBLE RABIES VACCINE IN MAIZE

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

Plants are natural bioreactors and are effective for production of recombinant proteins and antigens. There 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.

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

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

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

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.

prom. CMV Amp RV-gp Poli. A ORI Fodor I et al, Acta Vet Hung 2000, 48: 229 -326

CONSTRUCTION OF VECTORS MAR prom. Ubi Gen G Ter p. UCp. SS MAR Vector p. GHNC 5 Gen N 35 S Ca. MV 4. 9 Kb Ter Ca. MV

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

DETECTION OF PROTEIN, USING WESTERN BLOT 1 69 k. Da 2 3 4 5 6 7 65 k. Da Lane 1 -4, G protein expressed by transformed maize; lane 5, non transformed maize; lane 6, purified G protein; lane 7, MW.

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.

Survivorship of mice Survivorship in mice immunized with three different treatments and challenged intracraneally with 100 LD 50 of a vampire-bat strain.

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

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