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

Elizabeth Loza Rubio, NCVM-INIFAP, MexicoEdith Rojas Anaya NCVM-INIFAP Luis Gómez Nuñez NCVM-INIFAPTeresa Olivera Flores, UNAMMiguel Gómez-Lim, CINVESTAV

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.

POTENTIAL ADVANTAGES FOR THE PRODUCTION OF RECOMBINANT PROTEINS IN PLANTS

Plant systems are more economical than industrial facilities usingfermentation 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 pathogensor toxins are minimized

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

SOME ADVANTAGES OF ANTIGENS EXPRESSED IN TRANSGENIC PLANTS

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.

RV-gp PoliApromCMV

Fodor I et al, Acta Vet Hung 2000, 48:229-326

Amp ORI

Gen N35S CaMV Ter CaMV

4.9 Kb

MAR promUbi Gen G Ter pUCpSS MAR

Vector pGHNC5

CONSTRUCTION OF VECTORS

1 2 3 4 5 6

Figure 3. 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.

1231 bp20001200

800

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

1 2 3 4 5 6 7

Lane 1-4, G protein expressed by transformed maize;lane 5, non transformed maize; lane 6, purified G protein;

lane 7, MW.

69 kDa

DETECTION OF PROTEIN, USING WESTERN BLOT

65 kDa

0

0.5

1

1.5

2

2.5

1 2 3 4 5 6 1 2 3 4 5 6 1 2 3 4 5 6

Grupo 1 Grupo 2 Negativo

Mice groups

Ant

ibod

y tit

ers(

IU)

Group 1. Mice were immunized with a modified liverabies virus vaccine; Group 2. Adult mice were feed with 50g of G protein expressed in maize (orally); Group 3. Control non-transformed corn.

Virus neutralizing anti-rabies antibodiesin mice.

0

20

40

60

80

100

120

1 3 5 7 9 11 13 15 17 19 21 23 25 27 29

Days post-challenge

Surv

ival

(%

)

G1

G2

G3

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

Survivorship of mice

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

CONCLUSION

Merçi bien!

Thank you!

¡Gracias!Danke!

Спасибо

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