After mosquito inoculation of a human host, cellular entry likely resembles that of other flaviviruses,

whereby the virus enters skin cells through cellular receptors, enabling migration to the lymph nodes
and bloodstream.
Few studies have investigated the pathogenesis of Zika virus infection.
One study showed that human skin fibroblasts, keratinocytes, and immature dendritic cells
allow entry of Zika virus.[19]
Several entry and adhesion factors (e.g., AXL receptor tyrosine kinase) facilitate infection, and
cellular autophagy, needed for flaviviral replication, enhances Zika virus replication in skin
fibroblasts.[19]
After cellular entry, flaviviruses typically replicate within endoplasmic reticulum-derived vesicles.
However, Zika virus antigens were found exclusively in the nuclei of infected cells; this finding
suggests a location for replication that differs from that of other flaviviruses and merits further
investigation.

Aspects of Zika virus pathogenesis remain unclear. Zika virus's association with neurologic sequelae,
including potential neuropathophysiologic mechanisms, is being actively investigated.

Structure of ZIKV
Similar to other flaviviruses, ZIKV is an enveloped, icosahedral virus with a non-
segmented, single-stranded, positive-sense RNA genome, closely related to the
Spondweni virus.

The genome is 10,794 bases long, and has two non-coding flanking regions, the 5 NCR and
the 3 NCR.

It comprises a single long open reading frame: 5-C-prM-E-NS1-NS2A-NS2B-NS3-NS4A-

NS4B-NS5-3.

The NS1, NS3, and NS5 are large, highly-conserved proteins, while the NS2A, NS2B, NS4A,
and NS4B proteins are smaller, hydrophobic proteins.

The ORF codes for a polyprotein that subsequently cleaves into the capsid (C), precursor
membrane (prM), envelope (E), and non-structural proteins (NS).

The 5 end has a methylated nucleotide cap.

The non-polyadenylated 3 terminus has a looped structure.

The secondary structure leads to the formation of a sub-genomic flavivirus RNA (sfRNA),
which is essential for pathogenicity.

The E protein composes the majority of the virion surface and is involved with aspects of
replication, such as host cell binding and membrane fusion.

Cryo-electron microscopic studies showed ZIKV to be similar to other flaviviruses.

The envelope (E) protein is very similar to the West Nile virus, Japanese encephalitis virus,
and the dengue virus [13].

The virion is about 40 nm in diameter, having 5-10 nm surface projections arranged in an

icosahedral symmetry.

The ZIKV contains a nucleocapsid (approx. 25-30 nm in diameter), surrounded by a lipid

bilayer, containing the prM and E envelope proteins [13].

Replication of ZIKV
Using the PF-13 ZIKV strain isolated during an outbreak in French Polynesia, Hamel et al.
extensively studied the interaction of ZIKV in humans[14].
 They observed that fibroblast, keratinocytes, and immature dendritic cells are all
susceptible to ZIKV infection, and support viral replication.

Once the virus is deposited on the epidermis and dermis of the host, there is a rapid
increase in RNA copy numbers and ZIKV particles, which indicate active viral replication.

replicate initially in the dendritic cells at the site of infection where it then spreads to other areas of the body
via the bloodstream and lymphatic system. Viral replication typically occurs in the cytoplasm of infected
cells, but there is a study that shows ZIKV antigens can also be found in the nuclei of infected host cells

The ZIKV infection of the epidermal keratinocytes results in cell apoptosis, indicated by the
appearance of cytoplasmic vacuolation and the presence of pyknotic nuclei in the
stratum granulosum.

Inducing apoptotic cell death prevents the antiviral immune response, and increases the
dissemination of the virions.

The E protein is responsible for the fusion and entry of the virus through the host
membrane.

However, the exact mode of entry is not yet understood.

Post dissemination, the virus continues to spread to the lymph nodes and blood stream.

Although flaviviruses are generally known to replicate in the cytoplasm, ZIKA antigens have
also been detected in the nuclei of infected cells [15].

Zika virus life cycle

The life cycle of ZIKV is similar to other known flaviviruses.
Briefly, the E proteins are involved in the attachment of the virus to receptors on the host
membrane.
Subsequently, the virus gets internalised via endocytosis.
The viral RNA is released into the cytoplasm following fusion of the viral and host
membranes.
The ssRNA is then translated, and the resulting polyprotein further gets cleaved into
various structural and non-structural proteins (C, prM, E and NS proteins) [16].
Replication takes place on the ER surface. ZIKA antigens have also been detected in the
nuclei of infected cells
Transcription and replication of the dsDNA result in the formation of new viral mRNA and
ssRNA, respectively.
Following viral assembly at the ER, the virion budding utilises the host ESCRT (endosomal
sorting complexes required for transport) machinery to transport to the Golgi apparatus.
The prM protein is cleaved, and the virion is rendered mature.
Finally, the mature virus exits the cell via exocytosis [16].

Host Immune Response [14]

Following viral infection the hosts immune system recognises viral proteins and elicits an
antiviral response.

Previous studies on flaviviruses showed that both TLR3 and TLR7 (Toll-like receptors) are
able to recognise molecules on these pathogens.

Following recognition, the TLRs induce the host defense, initiating various signalling
pathways, which lead to the production of type I Interferons (IFNs), inflammatory
cytokines and chemokines.
 However, an in vitro study by Hamel et al on human fibroblast cell lines showed that ZIKV
infection only enhanced TLR3, but not TLR7.

The increased levels of TLR3 were accompanied with the production of IFN- and IFN- in
the infected cells, a corresponding increase in the transcriptional levels of interferon
regulatory factor 7 (IRF7), and the upregulation of expression of several IFN-stimulated
genes.

Simultaneously, there is an increase in the expression of two chemokines (CXCL10 and

CXCL11), which are known to play crucial roles in adaptive as well as innate immunity.
Upregulation of CCL5, another chemokine known for its antiviral activity, was also
observed.

As an antiviral host defence mechanism, the researchers found that both types I and II
interferons are capable of inhibiting virus replication.

The Hamel et al study also showed that ZIKV infection triggered autophagy in the infected
fibroblasts.

Generally, autophagy-mediated degradation of viral proteins prevents the replication of the

virus; however, some viruses, including ZIKV, utilize certain components of the autophagy
cycle, and promote viral replication and dissemination.

Electron microscopic studies also revealed membrane vesicles (70 100 nm) in close
association with the ER; this indicates that ZIKV replication occurs intimately with the
host cell membrane.

Reproductive Cycle of a Zika virus in a Host Cell

The reproductive cycle of ZIKV follows that of other known flaviviruses.

First, the virion attaches to the host cell membrane receptors via the envelope protein which induces
virion endocytosis.

Next, the virus membrane fuses with the endosomal membrane and the ssRNA genome of the virus is
released into the cytoplasm of the host cell.

It is then translated into a polyprotein that is subsequently cleaved to form all structural and non-
structural proteins.

Replication then takes place at intracellular compartments known as cytoplasmic viral factories in the
endoplasmic reticulum resulting in a dsRNA genome.

The dsRNA genome is then transcribed resulting in additional ssRNA genomes.

Assembly then occurs within the endoplasmic retiiculum and the new virions are transported to the
Golgi apparatus and then excreted into the intracellular space where the new virions can infect new
host cells. [6]