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
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