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Development of a vaccine to treat

dengue types and test their effect.


Present :
Miguel de Jesus Reyes Castillo
Miguel Angel Escalera Lugo
Yosselin Irazar Perez Rodriguez
Jonathan Silva Mendoza
Abdiel Eleuterio Guerrero
Introduction

Flaviviridae, ARN
Aedes aegypti
single chain +

Serotypes:
DEN-1, DEN-2,
DEN-3, DEN-4
At present it is a problem in several countries of Latin America because its
incidence has increased in the last decades. It has caused about 1181 deaths,
only in America.
Materials and methods
Obtaining of strains West pac 74
(DEN-1), S16803, (DEN-2), CH53489
(DEN-3), TVP360 (DEN-4)

Cells were trypsinized


Adaptation of VERO cells and subcultured
Growth medium (5
days)
Inoculation of the
virus
Growth medium
(5days)
Preparation of antigens
Virus concentration by tangential
flow ultrafiltration
Sorvall
RT6000B

Sucrose gradient ultracentrifugation

Beckman
Optima
Max-XP

Incubation for
Virus inactivation with formalin
approximately 15 days
Evaluation of
immunogenicity

Virus plaque reduction


neutralization (PRNT)
assay
Inoculation of vaccines in monkeys

1.- 15 macaques rhesus were randomly assigned to 3 groups of 5 individuals each.


2.- Each one received formulations of the vaccine or saline solution, with 30 days
difference.
3. One group received formulations of the vaccine with doses of antigen plus
adjuvant (aluminum hydroxide)
4.- Another group received the formulation of the vaccine without adjuvant
5. A negative control group received phosphate buffered saline (PBS)
6. The dengue virus (DEN-1-DEN-4) was inoculated in each monkey.
7.- Monkeys health status is monitored every day.
Adyuvant
the adjuvants are heterogeneous and nonspecific agents that associated
with the vaccine antigens induce a more powerful and prolonged immune
response
Bibliography

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Simmons M, Burgess T, Lynch J, Putnak R, 2010. Protection against dengue virus by non-
replicating and live attenuated vaccines used together in a prime boost vaccination strategy.
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Sun W, Nisalak A, Gettayacamin M, Eckels KH, Putnak JR, Vaughn DW, Innis BL, Thomas SJ,
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Stefan Fernandez, Stephen J. Thomas, Rafael De La Barrera, Rawiwan Im-erbsin, Richard G.
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Kenneth H. Eckels*, 2015. An Adjuvanted, Tetravalent Dengue Virus-Purified Inactivated
Vaccine Candidate Induces Long-Lasting and Protective Antibody Responses Against
Dengue Challenge in Rhesus Macaques