ZIKA Virus and Host Immunity

Yoes Prijatna Dachlan

Faculty of Medicine
Universitas Airlangga

Poltekes Semarang Nov/19/2016

 Zika is spread mostly by the bite of an infected Aedes species
mosquito (Ae. aegypti and Ae. albopictus). These mosquitoes
bite during the day
Zika can be passed from a pregnant woman to her fetus
Infection during pregnancy can cause certain birth defects
There is no vaccine or medicine for Zika
The association of Zika virus infection with Guillain-Barr

syndrome (GBS) and congenital birth defects (particularly

microcephaly) amid the ongoing outbreak of Zika virus
infection in Brazil is still under investigation

(CDC, 2016)
(Yoes Prijatna Dachlan, 2016)
 asymptomatic clinical courses
clinical resemblance to other infection
The global prevalence of Zika virus owing to
with other flaviviruses (dengue, chikungunya)
has not been widely reported
difficulty in confirming diagnosis

the incubation period : 3-12 days

In most cases, Zika virus infection
Epidemiology of causes a mild, self-limited illness
Owing to the mild nature of the disease
Zika infection > 80% of Zika virus infection cases likely
go unnoticed

Zika virus was first isolated in

1947 from the blood of a Rhesus
Currently, Zika virus is known to be widely
monkey (Macaca mulatta) in Zika
distributed outside of Africa. Outbreaks
forest, near Entebbe in Uganda
have been described previously in
(Dhurba Giri, 2016) and was
Micronesia and French Polynesia
reported in a human field worker
shortly thereafter (Bhagyashri D Navalkele BD, 2016)

(Yoes Prijatna Dachlan, 2016)

The directed lines connect the most probable sources and target
localities of viral lineages (shown by arrows), with widths
proportional to the posterior probabilities and values shown in
red. Only plausible routes with probabilities above 50% are
shown. The distinct introductions into Senegal and Cte
d'Ivoire were represented by different colors. The estimated
time to the most recent common ancestor of strains from
different countries are shown with 95% posterior time intervals
in parenthesis and could be interpreted as the oldest possible
year of introduction of that lineage at that locality. Letter
codes: UG Uganda CF Central African Republic DE
Dezidougou in Cte d'Ivoire SS Sokala-Sobara in Cte
d'Ivoire KE Kedougou in Senegal SA Saboya in Senegal
BA Bandia in Senegal DA Dakar in Senegal BF Burkina
Faso NG Nigeria MY Malaysia FM Yap Island in the
Federated States of Micronesia
 Transmission and Risks (1)
Through mosquito bites
Zika virus is transmitted to people primarily through the bite of
an infected Aedes species mosquito (Ae. aegypti and Ae. albopictus).
These are the same mosquitoes that spread dengue and chikungunya
viruses
These mosquitoes typically lay eggs in and near standing water
Mosquitoes that spread chikungunya, dengue, and Zika are aggressive
daytime biters, but they can also bite at night
Mosquitoes become infected when they feed on a person
infected with the virus. Infected mosquitoes can then spread the virus
to other people through bites
Through sex
It can be passed from a person with Zika before their symptoms start,
while they have symptoms, and after their symptoms end
Though not well documented, the virus may also be passed by a person
who carries the virus but never develops symptoms.
Zika can remain in semen longer than in other body fluids, including
vaginal fluids, urine, and blood
(Yoes Prijatna Dachlan, 2016)
From mother to child
A pregnant woman can pass Zika virus to her fetus during
pregnancy. Zika is a cause of microcephaly and other severe
fetal brain defects. We are studying the full range of other
potential health problems that Zika virus infection during
pregnancy may cause.
A pregnant woman already infected with Zika virus can pass
the virus to her fetus during the pregnancy or around the time
of birth.
To date, there are no reports of infants getting Zika virus
through breastfeeding
Through blood transfusion
There have been multiple reports of blood transfusion
transmission cases in Brazil. These reports are currently being
investigated
During the French Polynesian outbreak, 2.8% of blood donors
tested positive for Zika and in previous outbreaks, the virus
has been found in blood donors
(CDC, 2016)
 Transmission and Risks (2)

Through laboratory and healthcare setting exposure

Prior to the current outbreak, there were four reports of
laboratory acquired Zika virus infections, although the
route of transmission was not clearly established in all
cases.
As of June 15, 2016, there has been one reported case of
laboratory-acquired Zika virus disease in the United States.
To date, no cases of confirmed Zika virus transmission in
healthcare settings have been reported in the United States.
Recommendations are available for healthcare providers to
help prevent exposure to Zika virus in health care setting

Risks
Anyone who lives in or travels to an area where Zika virus is found and has not already been infected with Zika virus can get it from
mosquito bites. Once a person has been infected, he or she is likely to be protected from future infections

(CDC, 2016)
(Yoes Prijatna Dachlan, 2016)
 ZIKA Virus is an Emerging Global Pathogen
(Beckham JD; Pastula DM; Massey A; Tyler KL, 2016)
(Wiki,2016)
Zika virus (ZIKV) belongs to the Flavivirus genus of the Flaviviridae
family, which includes globally relevant arthropod-transmitted human
pathogens such as dengue (DENV), yellow fever (YFV), West Nile
(WNV), Japanese encephalitis (JEV), and tick-borne encephalitis
viruses (Lazear and Diamond, 2016, Pierson and Diamond, 2013).
Within the mosquito-borne clade of flaviviruses,
ZIKV is a member of the Spondweni group; both genetically and
serologically, ZIKV is related closely to the four serotypes of DENV
with approximately 43% amino acid identity and extensive antibody
cross-reactivity (Alkan et al., 2015, Lanciotti et al., 2008).
Mice sustained high viral loads in the brain and spinal cord, consistent
with evidence that ZIKV causes neurodevelopmental defects in human
fetuses.
( Lazear HM et al., 2016)
A Mouse Model of Zika Virus Pathogenesis
Zika virus is an icosahedral, enveloped, single-stranded RNA virus.[1]
The lipid envelope is covered with dense projections that consist of a
membrane and envelope glycoproteins
 Beraneka ragam organisme dan molekul- molekul yang dimilikinya
merupakan suatu ancaman konstan terhadap tubuh manusia

Mekanisme pertahanan manusia mewujudkan suatu proteksi dari serangan

penyakit dikenal dengan sebutan: imunitas (immunity)

Jejaring (network) terdiri dari sel, molekul, organ, dan jalur-jalur yang
bertanggung jawab atas imunitas tersebut kemudian membangun suatu
sistem disebut sistem imun (immune system). Jejaring tersebut
SISTEM IMUN bersifat dinamis dan dibawah kooordinasi yang tinggi serta kompleks

Sistem imun harus mampu membedakan organisme non-self (nonself

organism; mikroorganisme patogenik) dan molekul self yang merupakan
bagian dari tubuh manusia

Respons terkoordinasi dan kolektif terhadap substansi asing (nonself

organism) disebut respons imun (immune reponse)

Mekanisme defens: identifikasi (rekognisi/mengenal) substansi asing

menetralisasi dan menghancurkan substansi asing
(Yoes Prijatna Dachlan, 2016)
 Cells, Organs, Infectious
and disease

Microenvironment foreign substances Non

infectious
of the Immune System disease
Immunity protection from disease

Organs, molecules, cells, pathways

constitute
Network (interconnected) responsible for the immune system
immunity

accurate
harmony collective and coordinated response immune response
collaborative

(Yoes Prijatna Dachlan, 2016)

Sistem limfatik: terdiri atas pembuluh limfatik, kelenjar getah bening
(lymph nodes), dan organ limfatik, termasuk limpa dan kelenjar timus

(Yoes Prijatna Dachlan, 2016)

 Innate and adaptive immune cells of the human body
All cells are derived from self-renewing haematopoietic stem cells in the bone marrow, and
they arise from myeloid or lymphoid progenitors

(Lamb TJ, 2012)

(Yoes Prijatna Dachlan, 2016)
 Prinsip utama Respons Imun
Eliminasi mikroba melalui sistem imun innate dengan
menggunakan mekanisme non-spesifik
Isyarat diberikan oleh sistem imun innate kepada sistem
imun adaptive untuk menyiapkan respons imun dan
jenisnya
Sel-sel sistem imun adaptive melaksanakan rekognisi
spesifik terhadap Ag dan kemudian memobilisasi
mekanisme yang berpotensi untuk mengeliminasi
mikroorganisme tsb
Sistem imun membuat memori atas respons yang
terdahulu
Tolerans terhadap self-antigens

(Yoes Prijatna Dachlan, 2016) (Paul WE., 2008)

 Immune System

Adaptive
Innate
(Spesifik)
(Non-spesifik)
2nd of defence
1st of defence Melindungi dari paparan ulang

komponen seluler komponen humoral komponen seluler komponen humoral

(Gene Mayer, Ph.D, 2011)

(Yoes Prijatna Dachlan, 2016)

 Innate Adaptive
Non-specific Immunity Specific Immunity
Response is antigen-independent Response is antigen-dependent
There is immediate maximal response There is a lag time between exposure and maximal
response
Not antigen-specific Antigen-specific
Exposure results in no immunologic memory Exposure

(Gene Mayer, Ph.D, 2011)

(Yoes Prijatna Dachlan, 2016)

 Mechanic and
chemical barrier
M Neu

B1 B T NK
cells cells

DC
M
Innate immune
system

T
T cells
cells
B2 B
cells

Adaptive immune
system

(Yoes Prijatna Dachlan,2016)

 Innate immune mechanism Cell types Function

Barrier defence Epithelial cells Prevent pathogen entry

Humoral Soluble factors Opsonisation

Complement proteins Cytotoxicity
Acute phase proteins Sistemic effects
Antibodies

Phagocytosis Macrophages (M) Engulfment of foreign particles

Neutrophils and cell lysis

Cytotoxicity NK cells
Neutrophils Cell lysis
Eosinophils

Cell signaling All innate immune cells Activation of innate and adaptive
immune responses

Antigen presentation Macrophages (M) Activation of T and B cells

Dendritic cells

(Yoes Prijatna Dachlan, 2016) (William AE et al., 2012)

 Respons mencegah
mengendalikan infeksi
dini
eliminasi

eliminasi
Immune sel-sel yang rusak
mechanisms mengawali proses
perbaikan jaringan

PAMPs
Recognisi
FUNGSI mikroba DAMPs (Shields AM et.al., 2011)
INNATE RAMPs
terhadap
MIKROBA Stimulasi respons imun adaptive ;
Mempengaruhi karakter respons adaptive
menyusun perlawanan efektip dan optimal
terhadap berbagai jenis mikroba

Inflammation

Jenis respons

Antiviral defense

(Yoes Prijatna Dachlan 2015) (ABBAS, 8th ed., 2015)

 Rekognisi mikroba

TLRs permukaan sel

Toll-like receptors Endosomes
NLR
Nucleotide binding
oligomerization domains
(NODs)-like receptors
PAMPs
CLRs rekognisi
PRRs
C-type lectin receptors: Struktur/Komponen/Produk
Pattern recognition Dectin-1, Dectin-2, DC-
receptors mikroba
SIGN

RLR
RIG-I-like receptors:
mediator inflamatori
RIG-I, MDA5

Intra-cytosolic DNA
sensors
eliminasi koordinasi
patogen invasiv pengembangan
respons imun
PAMPs: Pathogen-associated molecular patterns
adaptive

(Yoes Prijatna Dachlan, 2015) (Mogensen TH, 2009; Savva A,Roger T, 2013)
 PAMPs
Molekul yang karakteristik pada
setiap klas patogen

Respons Imun Host factors sebagai sinyal danger

Innate recognisi berada di lokasi yang menyimpang
PRRs (e.g. endosomal DNA)
kompleks molekuler abnormal
(e.g. self DNA autoab complexes

other types of cellular stress

Resolusi Inflammatory disease/

infeksi autoimmunity
(Yoes Prijatna Dachlan, 2014)
ZIKV Genomic Evolution and Pathogenesis
Virus in the 20th century: analysis and comparison with past isolates. A recent analysis of the molecular evolution of ZIKV
isolates ZIKV likely first emerged in Uganda around 1920 followed by 2 independent ZIKV introductions into West and
Central Africa from the eastern portion of the continent a third introduction event into Malaysia around 1945
During its emergence, ZIKV underwent 13 recombination events, which is an unusual feature for a flavivirus since these viruses
are often genetically restricted by the need to replicate in evolutionarily disparate invertebrate (mosquitoes) and vertebrate
hosts
The unusual evolutionary plasticity of the ZIKV genome is likely due in part to vertebrate host preferences for the virus,which
may contribute to ZIKV genomic adaptation to specific vectorvertebrate host environments
This likely resulted in an increased frequency of ZIKV activity every1 to 2 years when compared with other arboviruses,which
exhibit an activity frequency of 5 to 8 years for dengue virus and yellow fever virus
The unique genomic features of ZIKV likely contributed to the rapid emergence of the virus over a very large geographic area.

(Beckham JD; Pastula DM; Massey A; Tyler KL, 2016)

(Yoes Prijatna Dachlan, 2016)
 Innate immune responses
ZIKV

Recognition
Entry route
of PAMPs by PRRs

stimulates
an intracellular
signaling
cascade

Dermal Fibroblasts and

Epidermal Keratinocytes

(Beckham JD; Pastula DM; Massey A; Tyler KL, 2016)

(Yoes Prijatna Dachlan, 2016)

 Biology of Zika Virus Infection in Human Skin Cells

ZIKV is transmitted
by the Aedes mosquito

Aedes deposits the virus

in the epidermis and dermis
the epidermis and dermis
Several entry, including DC-SIGN, AXL, Tyro3,
and, to a lesser extent, TIM-1, permitted ZIKV entry
Human keratinocytes
cytoplasmic vacuolation
superficial subcorneal edema
Infection
of skin fibroblasts
resulted in
the presence of high
RNA copy numbers

indicating

Partial degradation of cytoplasm by the the virus uses autophagy active viral replication in the
cell itself cellular apoptosis infected cells.
(Rodolphe Hamel et al., 2015)

boosts dissemination
(Actualite scientifique, No. 483, 2015)
(Yoes Prijatna Dachlan, 2016)
How the zika virus infects human cells
 ZIKV induces an innate antiviral response in primary
human skin fibroblasts

ZIKA (Beckham JD; Pastula DM; Massey A; Tyler KL, 2016)

(McCartney SA, 2010)
(PAMPs)

short dsRNAs
long dsRNAs

The induction of TLR3 expression is rapid, Fibroblast surface

whereas that of RIG-I and MDA-5 is delayed.

RIG I MDA5
Endosome (Slater L et al., 2010)
ZIKV-infected fibroblasts Co-ordinated Role of TLR3, RIG-I and MDA5
in Bronchial Epithelium
 PAMPs PRRs
(Pathogen-associated (Pattern Recognition Receptors)
Molecular Patterns)

TLR3 TLR3

adaptor TRIF

IKKa b IRF-3 (INF Regulatory Factor 3)

IRF-7
IFN-
NF-B production
type I IFNs
proinflammatory Three classes of pattern-recognition receptors (PRRs) for RNA virus recognition
cytokines

Osamu Takeuchi1, Shizuo Akira

Immunological Reviews 2009
(Yoes Prijatna Dachlan, 2016)
 Interferon (IFN)s and innate immune responses

Type I and type II IFNs are known to be important for control of other flavivirus infections
Replication of ZIKV in human fibroblasts is inhibited by both types of IFNs (type I and type II; in the
murine model)
The dendritic cell (DC) is one of the main target cells during infection by many flaviviruses, such as
DNEV, WNV and presumably the most potent antigen presenting cell (APC)s
No "cytokine storm effects" were observed in the serum during the acute phase of ZIKV patients with
more pronounced elevation of chemokines than inflammatory cytokines

( Wang T, Vasilakis N, Weaver SC, 2016)

Zika virus and host immunity

(Yoes Prijatna Dachlan, 2016)

 Provide partial protection in adult and infant mice against lethal
ZIKV infection

The extensive cross-reactivity of patient serum antibodies

against closely related flaviviruses such as DENV and YFV
do not seem to confer cross protection against ZIKV disease

Antibody-dependent enhancement (ADE) occurs through Fc receptors,

which bind the back ends of antibodies
Fc receptors are found on several
cell types, including macrophages
Adaptive immune responses (1) and placental epithelial cells.
They internalize antibodies and may
also internalize partially neutralized
viral particles bound by those
antibodies

Innate and adaptive immune response to ZIKV infection:

Correlates of risk? Correlates of protection?
The protective immune response is Abs or cell-mediated?
Does infection lead to lifelong protection?
Is cross-immunity conferred by infection by other flaviviruses
(implications for the clinical evaluation of vaccine candidates)?
(GloPID-R, 2016)
(Yoes Prijatna Dachlan, 2016)
 Although cross immunity does not inhibit related virus infection,
it may reduce the severity of infection to some extent
e.g. Rhesus monkeys immunized with ZIKV had a reduced
viremia upon a subsequent challenge with YFV

During WNV infection, T cells contribute to control of viral

dissemination and ultimate clearance of the virus

In a recent report, analysis of serum cytokine levels of travelers

Adaptive immune responses (2) returning from Asia, the Pacific and Brazil revealed elevated
A P
Th1 (IL-2),
C H
Th2 (IL-4, IL-13),
U A
Th17 (IL-17)
T S
Th9 (IL-9)
E E
during the acute phase,
but normal levels of these cytokines in the recovery phase

polyfunctional T cell responses are induced upon ZIKV infection

( Wang T, Vasilakis N, Weaver SC, 2016)

(Yoes Prijatna Dachlan, 2016)
 microbe
Antibody-dependent
enhancement (ADE) occurs
through Fc receptors, which bind the
back ends of antibodies
antigen Fc receptors are found on several
cell types, including macrophages
Ag-binding site
epitope and placental epithelial cells.
They internalize antibodies and may
also internalize partially neutralized
viral particles bound by those
Fab region antibodies
( Keener AB, 2016)
Fc region Antibody (Ab) Zika and Dengue Immunity:
A Complex Relationship

Fc receptor

phagocyte (Male, Brostoff, Roth, Roitt, 2006)

(Yoes Prijatna Dachlan, 2016)

 The Structure of the Human Placenta and the Role of IFN1 in Protecting against ZIKV Infection
The intrauterine environment during
human pregnancy. Embryonic
structures include the villous tree of
the human hemochorial placenta and
the umbilical cord, which transfers
blood between the placenta and the
fetus
(Yoes Prijatna Dachlan, 2016)
A single placental villus. Extravillous
trophoblasts invade and anchor the
placenta to the maternal decidua and to the
inner third of the myometrium
Multinucleated syncytiotrophoblasts
overlie the surfaces of the villous tree and
are in direct contact with maternal blood,
which fills the intervillous space (IVS)
once the placenta is fully formed
Mononuclear cytotrophoblasts are
subjacent to the syncytiotrophoblasts
The syncytiotrophoblasts release IFN
1 that can act in both autocrine and
paracrine manners to induce ISGs,
which protect against ZIKV and other
viral infections.
The paracrine function of IFNl could
work locally within the direct
maternal-fetal compartment or might
circulate
more systemically to act on other
maternal target cells
(Bayer A et al. Cell Host & Microbe 19, 2016)
 Placental trophoblast IFN production therefore is probably an important factor in limiting transplacental viral
and possibly bacterial spread from mother to fetus

The human placenta function as an immunologic barrier between the maternal and fetal circulation,
preventing potentially destructive maternal immune response from damaging semiallogeneic fetus. Local
immunosuppression has been one of the mean hypothesized to prevent the sensitization of the maternal
immune system to paternal alloantigens and development of subsequent effector functions, possibly leading
to the survival of the fetus. The ability of the trophoblast IFNs to suppress T- and B lymphocyte proliferation
may be one of the factors contributing to the immunosuppression at the feto-placental unit so that maternal
immune responses do not destroy pregnancy
The trophoblast IFNs may prevent virus infection and spreading by activating immunoreactive NK cells to lyse
virus-infected cell during pregnancy

(Yoes Prijatna Dachlan, 2016) (Aboagye-Mathiesen G, 1994)

 Images of the fetus brain at 36 weeks of pregnancy revealed
the brain was filled with fluid, which resulted in the childs
head appearing to be normal-sized.
the baby was also missing parts of the nervous system
including sections of the brain stem, the spinal cord and the
midbrain, which controls eye movements and processes
information from the eyes and ears.
the brain stopped growing in early development
the skull collapsing
skull was flattened with extra skin around the head

(Janice Williams, 2016)

Zika Virus Symptoms: New Pictures Show Most Severe Brain Damage In Infected
(Yoes Prijatna Dachlan, 2016) Fetuses
 ZIKA Virus Animal Models
Suitability for Vaccine Testing

(NIH, 2016)
(Yoes Prijatna Dachlan, 2016)
TERIMAKASIH

Faculty of Medicine
Universitas Airlangga
Surabaya, Indonesia