Diapositiva 1

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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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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.
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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
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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
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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
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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.
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promCMV
RV-gp
PoliA
Amp
ORI
pWR-21 (5610 PB)
Fodor I et al, Acta Vet Hung 2000, 48229-326
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CONSTRUCTION OF VECTORS
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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
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DETECTION OF PROTEIN, USING WESTERN BLOT
1 2 3 4 5 6
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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
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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
maizelane 5, non transformed maizepurified G
protein lane 7, MW.
Figure 4. Detection of G protein, using WB. Lane
1-4, G protein expressed by transformed
maizelane 5, non transformed maizepurified G
protein lane 7, MW.
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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.
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Survivorship of mice
Survivorship in mice immunized with
three different treatments and challenged
intracraneally with 100 LD50 of a vampire-bat
strain.
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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
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Merçi bien!
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Danke!