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• PRESENTERS:
• AYESHA IRFAN
• HADIA LATIF KIYANI
• RUMAISA ASIF
• IQRA HOORAIN
• VANEEZA ARSHAD
1
GenomeIntroduction
DENGUE VIRUS:
The genome of virus is single positive stranded
RNA.
Belongs to the family Flaviviridae and genus
Flavivirus
Arbovirus
2
DENGUE VIRUS:
• Virus is spread by mosquito of genus, Aedes.
• Most important vector of dengue virus is, Aedes
aegypti.
• Aedes is found in tropical and sub-tropical regions
of world.
• The mosquito is small dark colored with typical
white marking on body.
3
Structure
Roughly spherical Diameter: 50nm
Viral genome is surrounded by capsid (C)
protein
Capsid is surrounded by viral envelope
Embedded in viral envelope are 180
copies of the E & M proteins by a short
transmembrane segment that span
through the lipid bilayer
4
Serotypes
SEROTYPES
OF DENGUE
VIRUS
DEN-1 DEN-2
DEN-3 DEN-4
5
HISTORY
• First probable record found in Chinese medical encyclopedia-
“water poison by a flying insect”
• First endemic occurred simultaneously in Asia and Africa.
• First confirmed case was reported in 1789.
• The virus was first isolated in 1943 by Kimura and Hotta.
• The virus gets its name from DINGA word of Spanish origin.
• The word is derived from a phrase of Swahili language meaning :
“Cramp seizure by an evil spirit”
6
Epidemiology:
7
According to ECDC-2019
Epidemiology:
AMERICA:
• Belize
• Honduras
• Brazil
Reported
cases:
2,848,381
AFRICA:
• Benin
• Sudan
• Tanzania
Reported
cases:
24,983
ASIA:
• Japan
• Maldives
• Bangladesh
• Pakistan
• India
Reported cases:
1,51,904
AUSTRALIA:
In Australia
the reported
cases are:
1268
8
AMERICA:
• Belize
• Honduras
• Brazil
Reported
cases:
2,848,381
AFRICA:
• Benin
• Sudan
• Tanzania
Reported
cases:
24,983
Epidemiology:
DENGUE OUT BREAK IN
PAKISTAN 2019:
9
Genome and Transmission
of Dengue Virus
Presented By: Hadia Latif Kiyani
10
Genome
Single stranded +sense RNA
It can directly translated into proteins
Consists of 10,700 (~11kb) nucleotides and encodes 3,411 amino acids long
precursor polyprotein
The genome is a single long open reading frame (ORF), flanked by 5’
and 3' untranslated regions (UTRs)
Out of these proteins 3 are structural and 7 are non-structural proteins
Polyprotein is processed co- and post-translationally, by cellular and viral proteases,
to produce 10 mature viral proteins
11
Genome
12
1 C (Capsid) protein: Viral RNA packaging
2 prM: Precursor for M (membrane) protein
3 E protein: Envelope glycoprotein, The job of the
envelope protein is to attach to a cell surface
and begin the process of infection (receptor
binding and fusion)
Structural proteins
13
Non-structural
proteins
Non structural
proteins
Function
• Require for anchoring of viral
replication complex
• Cofactor for NS3 serine protease
• Serine protease/helicase/NTPase
• Inhibition of interferon signal
transduction
• RNA dependent RNA polymerase
(RdRp); methyl transferase
• NS1
• NS2B
• NS3
• NS4B
• NS5
14
Transmission of
Dengue Virus
Transmission of Dengue
Virus
15
Transmission
Vector Transmission
16
Transmission
In rare events, dengue can be transmitted
Organ transplantations
Blood transfusions from infected donors
An infected pregnant mother can transmit the
dengue virus to her fetus
Non Vector Transmission
17
Life cycle of dengue virus
Presented By: RUMAISA ASIF
Virus Attachment
• DENV replicate in a variety of cells( liver, spleen, lymph node, kidney and other
organs)
• Doesnot use specific receptors but binds to diverse molecules
16Alen, M.M. and Schols, D., 2012. Dengue virus entry as target for antiviral therapy. Journal of tropical medicine, 2012.
Entry
• Clatherine mediated endocytosis
• Endosome formed
Fusion
• Low pH in the endosome
• Envelope glycoproteins form a trimeric knob like str
• Domain II of E protein project outwardly
• Hydrophobic fusion loop in domain II
• inserted into endosomal membrane
Uncoating
17
Translation of viral genome
• Host ribosomal machinery utilized
• Single polypeptide formed
• Cleavage of polypeptide
• Viral proteins synthesized
21
Replication
• Replication complex formed
• Cyclization of the genome occurs
• -RNA strand synthesized first
• +RNA strand synthesized from - strand
RC
Viral proteins
Viral genome
Host cell factors
22
Assembly and packaging
• At ER membrane
• Neucleocapsid enveloped by ER membrane
• Immature virion is formed
Maturation and Release
• In trans Golgi apparatus(TGA)
• prM cap the fusion peptide(at domain II)
• prM cleaved into M protein and pr-peptide by furin
• Pr-peptide disassociated upon virus release via exocytosis
24
25International Journal of Medical Sciences 14: 1342 image No. 003
Pathogenesis
classification of dengue
Presented by Iqra Hoorain
26
Dengue fever
dengue
hemorrhagic
fever
Dengue shock
syndrome
Infection
Types
27
Phases of
infection
28
 The DENV are closely related, antigenically
distinct
 increases the risk of severe dengue
 high potential for mutation
 DENV strains vary in their ability to infect
different cell types or cause severe disease
 RISK FACTORS INCLUDE both host and viral
factors
 Specific virus serotypes and genotypes
replicate more readily in specific population
groups
Serotypes and
viral virulence
DENV1 DENV2 DENV3 DENV4
29
Pathogenesis
Original
antigenic sin
Antibody-
dependent
enhancement
Autoimmunity
Cytokine
storm
30
Infection with
a single virus
is followed by
up to three
outcomes:
durable
protection
against
infection
with a strain
of the same
DENV,
brief
protection
against
infection or
disease with a
different
dengue
serotype
infection with
a different
dengue
serotype that
may result in
severe
disease
31
proliferation of cross-reacting
memory-cells induced by the primary-
infection
lower-affinity of binding with the
infected-macrophages, not being as
effective in clearing secondary-
infections
boosting the immune-pathology and
damage to cells
32
Antibody-
dependent
enhancement
Minor antigenic-differences between the
serotypes, ineffective neutralization.
Antibodies bind heterologous DENV via
Fcγ receptor expressed on target cells
High viremia.
High amount of cytokines being released.
Induce capillary endothelial pathology
and vascular leakage, leading dengue
shock syndrome.
33
34
Autoimmunity
• Antibodies produced during a DENV cross-react with some
self-antigen.
• NS1 is the only NS protein that can be released into the
bloodstream.
• The hypothesis is that anti-DV NS1 is involved in the
pathogenesis of DHF/DSS
• Antibodies recognizing a linear epitope in the E protein to
bind human plasminogen and inhibit plasmin activity.
35
Vaccines treatment prevention
Presented By: Vaneeza Arshad
DIAGNOSTIC TESTS
1. Viral isolation
2. Antigen detection
3. Genome detection
4. Antibody detection
n.
tient
ltured
A
B
s
ies
hase
t)
c
D
us
35
TREATMENT
Prevent
dehydration
painkillers
NSAIDS
IV Fluids
38
CONTROL
STRATEGIES
Chemical
control• Use of insecticides
• Use of pheromones as
“attract ands kill”
39
VACCINATION
• Dengavaxia
• First licensed in Mexico Dec 2015
• Administered as a 3 dose series
• 9-45 years
• Now licensed in 20 countries
• Only given to people with confirmed prior dengue virus infection
• Other vaccines under trial
DENVax (phase II trials)
TDENV PIV
V-180 ( phase I stage)
40
PREVENTION
• Clothing
• Mosquito repellents
• Mosquito traps and nets
• Avoid scents
• Timing
• Stagnant water
41
42
References
1. WHO. Dengue and severe dengue; 2017.
2. Bains S. Severe dengue infection. Medscape; 2017.
3. WHO. Dengue control; 2018.
4. CLINICAL MICROBIOLOGY REVIEWS, Oct. 2009, p. 564–581
5. Hidari, K.I. and Suzuki, T., 2011. Dengue virus receptor. Tropical medicine and
health, 39(4SUPPLEMENT), pp.S37-S43.
6. Hung, S.L., Lee, P.L., Chen, H.W., Chen, L.K., Kao, C.L. and King, C.C., 1999. Analysis of
the steps involved in dengue virus entry into host cells. Virology, 257(1), pp.156-167.
7. International Journal of Medical Sciences 14: 1342 image No. 003
8. Clyde, K., Kyle, J.L. and Harris, E., 2006. Recent advances in deciphering viral and host
determinants of dengue virus replication and pathogenesis. Journal of virology, 80(23),
pp.11418-11431.
9. Cruz-Oliveira, C., Freire, J.M., Conceição, T.M., Higa, L.M., Castanho, M.A. and Da Poian,
A.T., 2015. Receptors and routes of dengue virus entry into the host cells. FEMS
microbiology reviews, 39(2), pp.155-170.
THANKYOU
43

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Dengue virus

  • 1. • PRESENTERS: • AYESHA IRFAN • HADIA LATIF KIYANI • RUMAISA ASIF • IQRA HOORAIN • VANEEZA ARSHAD 1
  • 2. GenomeIntroduction DENGUE VIRUS: The genome of virus is single positive stranded RNA. Belongs to the family Flaviviridae and genus Flavivirus Arbovirus 2
  • 3. DENGUE VIRUS: • Virus is spread by mosquito of genus, Aedes. • Most important vector of dengue virus is, Aedes aegypti. • Aedes is found in tropical and sub-tropical regions of world. • The mosquito is small dark colored with typical white marking on body. 3
  • 4. Structure Roughly spherical Diameter: 50nm Viral genome is surrounded by capsid (C) protein Capsid is surrounded by viral envelope Embedded in viral envelope are 180 copies of the E & M proteins by a short transmembrane segment that span through the lipid bilayer 4
  • 6. HISTORY • First probable record found in Chinese medical encyclopedia- “water poison by a flying insect” • First endemic occurred simultaneously in Asia and Africa. • First confirmed case was reported in 1789. • The virus was first isolated in 1943 by Kimura and Hotta. • The virus gets its name from DINGA word of Spanish origin. • The word is derived from a phrase of Swahili language meaning : “Cramp seizure by an evil spirit” 6
  • 8. Epidemiology: AMERICA: • Belize • Honduras • Brazil Reported cases: 2,848,381 AFRICA: • Benin • Sudan • Tanzania Reported cases: 24,983 ASIA: • Japan • Maldives • Bangladesh • Pakistan • India Reported cases: 1,51,904 AUSTRALIA: In Australia the reported cases are: 1268 8 AMERICA: • Belize • Honduras • Brazil Reported cases: 2,848,381 AFRICA: • Benin • Sudan • Tanzania Reported cases: 24,983
  • 9. Epidemiology: DENGUE OUT BREAK IN PAKISTAN 2019: 9
  • 10. Genome and Transmission of Dengue Virus Presented By: Hadia Latif Kiyani 10
  • 11. Genome Single stranded +sense RNA It can directly translated into proteins Consists of 10,700 (~11kb) nucleotides and encodes 3,411 amino acids long precursor polyprotein The genome is a single long open reading frame (ORF), flanked by 5’ and 3' untranslated regions (UTRs) Out of these proteins 3 are structural and 7 are non-structural proteins Polyprotein is processed co- and post-translationally, by cellular and viral proteases, to produce 10 mature viral proteins 11
  • 13. 1 C (Capsid) protein: Viral RNA packaging 2 prM: Precursor for M (membrane) protein 3 E protein: Envelope glycoprotein, The job of the envelope protein is to attach to a cell surface and begin the process of infection (receptor binding and fusion) Structural proteins 13
  • 14. Non-structural proteins Non structural proteins Function • Require for anchoring of viral replication complex • Cofactor for NS3 serine protease • Serine protease/helicase/NTPase • Inhibition of interferon signal transduction • RNA dependent RNA polymerase (RdRp); methyl transferase • NS1 • NS2B • NS3 • NS4B • NS5 14
  • 17. Transmission In rare events, dengue can be transmitted Organ transplantations Blood transfusions from infected donors An infected pregnant mother can transmit the dengue virus to her fetus Non Vector Transmission 17
  • 18. Life cycle of dengue virus Presented By: RUMAISA ASIF
  • 19. Virus Attachment • DENV replicate in a variety of cells( liver, spleen, lymph node, kidney and other organs) • Doesnot use specific receptors but binds to diverse molecules 16Alen, M.M. and Schols, D., 2012. Dengue virus entry as target for antiviral therapy. Journal of tropical medicine, 2012.
  • 20. Entry • Clatherine mediated endocytosis • Endosome formed Fusion • Low pH in the endosome • Envelope glycoproteins form a trimeric knob like str • Domain II of E protein project outwardly • Hydrophobic fusion loop in domain II • inserted into endosomal membrane Uncoating 17
  • 21. Translation of viral genome • Host ribosomal machinery utilized • Single polypeptide formed • Cleavage of polypeptide • Viral proteins synthesized 21
  • 22. Replication • Replication complex formed • Cyclization of the genome occurs • -RNA strand synthesized first • +RNA strand synthesized from - strand RC Viral proteins Viral genome Host cell factors 22
  • 23. Assembly and packaging • At ER membrane • Neucleocapsid enveloped by ER membrane • Immature virion is formed
  • 24. Maturation and Release • In trans Golgi apparatus(TGA) • prM cap the fusion peptide(at domain II) • prM cleaved into M protein and pr-peptide by furin • Pr-peptide disassociated upon virus release via exocytosis 24
  • 25. 25International Journal of Medical Sciences 14: 1342 image No. 003
  • 29.  The DENV are closely related, antigenically distinct  increases the risk of severe dengue  high potential for mutation  DENV strains vary in their ability to infect different cell types or cause severe disease  RISK FACTORS INCLUDE both host and viral factors  Specific virus serotypes and genotypes replicate more readily in specific population groups Serotypes and viral virulence DENV1 DENV2 DENV3 DENV4 29
  • 31. Infection with a single virus is followed by up to three outcomes: durable protection against infection with a strain of the same DENV, brief protection against infection or disease with a different dengue serotype infection with a different dengue serotype that may result in severe disease 31
  • 32. proliferation of cross-reacting memory-cells induced by the primary- infection lower-affinity of binding with the infected-macrophages, not being as effective in clearing secondary- infections boosting the immune-pathology and damage to cells 32
  • 33. Antibody- dependent enhancement Minor antigenic-differences between the serotypes, ineffective neutralization. Antibodies bind heterologous DENV via Fcγ receptor expressed on target cells High viremia. High amount of cytokines being released. Induce capillary endothelial pathology and vascular leakage, leading dengue shock syndrome. 33
  • 34. 34
  • 35. Autoimmunity • Antibodies produced during a DENV cross-react with some self-antigen. • NS1 is the only NS protein that can be released into the bloodstream. • The hypothesis is that anti-DV NS1 is involved in the pathogenesis of DHF/DSS • Antibodies recognizing a linear epitope in the E protein to bind human plasminogen and inhibit plasmin activity. 35
  • 37. DIAGNOSTIC TESTS 1. Viral isolation 2. Antigen detection 3. Genome detection 4. Antibody detection n. tient ltured A B s ies hase t) c D us 35
  • 39. CONTROL STRATEGIES Chemical control• Use of insecticides • Use of pheromones as “attract ands kill” 39
  • 40. VACCINATION • Dengavaxia • First licensed in Mexico Dec 2015 • Administered as a 3 dose series • 9-45 years • Now licensed in 20 countries • Only given to people with confirmed prior dengue virus infection • Other vaccines under trial DENVax (phase II trials) TDENV PIV V-180 ( phase I stage) 40
  • 41. PREVENTION • Clothing • Mosquito repellents • Mosquito traps and nets • Avoid scents • Timing • Stagnant water 41
  • 42. 42 References 1. WHO. Dengue and severe dengue; 2017. 2. Bains S. Severe dengue infection. Medscape; 2017. 3. WHO. Dengue control; 2018. 4. CLINICAL MICROBIOLOGY REVIEWS, Oct. 2009, p. 564–581 5. Hidari, K.I. and Suzuki, T., 2011. Dengue virus receptor. Tropical medicine and health, 39(4SUPPLEMENT), pp.S37-S43. 6. Hung, S.L., Lee, P.L., Chen, H.W., Chen, L.K., Kao, C.L. and King, C.C., 1999. Analysis of the steps involved in dengue virus entry into host cells. Virology, 257(1), pp.156-167. 7. International Journal of Medical Sciences 14: 1342 image No. 003 8. Clyde, K., Kyle, J.L. and Harris, E., 2006. Recent advances in deciphering viral and host determinants of dengue virus replication and pathogenesis. Journal of virology, 80(23), pp.11418-11431. 9. Cruz-Oliveira, C., Freire, J.M., Conceição, T.M., Higa, L.M., Castanho, M.A. and Da Poian, A.T., 2015. Receptors and routes of dengue virus entry into the host cells. FEMS microbiology reviews, 39(2), pp.155-170.