Leading Edge
Select
Zika Virus on the Move
There is great urgency to understand Zika virus as it con-
tinues its spread throughout the Americas and in tropical re-
gions around the globe. What do we actually know about its
biology? And what basic research questions need to be
answered to counter its spread and effects?
The essentials are this: Zika falls in the category of positive
single-strand RNA virus and has a genome of 10 kb, which
encodes three structural proteins (capsid, premembrane/
membrane, and envelope) and seven nonstructural proteins.
By sequence homology, Zika is similar to other recently
emerging pathogens, West Nile, chikungunya, and dengue
viruses, and like these, it is also spread by the Aedes mos-
quito. Although the virus was first noted in Uganda in 1947,
the strains causing the current outbreak are part of the Asian
lineage, closely related to that which caused the 2007
epidemic in French Polynesia (Enfissi et al., 2016). This was
the first outbreak outside of Asia and Africa and was charac-
terized by a significant incidence of neurological symptoms,
including Guillain-Barre syndrome. It is unclear if the virus has
undergone changes that have accelerated its spread or that
contributed to its neurological sequelae, though one early
report proposes that codon usage for the nonstructural 1
(NS1) gene in the Asian lineage may be better optimized for
expression in humans (Freire et al., 2015). Although many
cases of Zika are asymptomatic and the most common
symptoms of infection are rash, fever, and joint pain, the
recent outbreak has caused alarm because it has been
associated with a significant increase in microcephaly in
newborns.
Lets start with the basicshow does Zika virus get first
into human cells? Hamel et al. (2015) recently explored this
question by exposing Zika virus to human skin cells (from
adult biopsies and neonatal foreskins) to simulate what hap-
pens after exposure to an infected mosquito. Similar to
dengue virus, the authors show that Zika appears to use
the C-type lectin receptor DC-SIGN and members of the
TIM and TAM families of phosphatidylserine receptors on
host cell surface to gain access to the cytoplasm via recep-
tor-mediated endocytosis. Their findings indicate that dermal
Image from iStock.com/doug4537.
fibroblast, keratinocytes, and immature dendritic cells can all
be readily infected and support viral replication. Skin den-
dritic cells in particular look to be an important Zika target
since they have also been proposed to facilitate the spread
of dengue virus. Assessment of other vulnerable cell types
awaits further investigation.
Hamel et al. also shed light on host-virus interactions that
shape the course of infection. In infected primary human fi-
broblasts, the virus triggers an innate immune response,
including expression of interferons, and both type I and
type 2 interferons inhibit viral replication. At the molecular
level TLR3, which recognizes double-stranded RNA appears
to be particularly central to the innate immune response to
Zika infection. This is reminiscent of other flaviviruses that
have been studied. At the cellular level the virus induces au-
tophagosome formation to promote replication and may
trigger apoptosis to foster viral dissemination. In broad
strokes, these cellular events appear similar to what is known
about dengue infection. Future work will delineate the extent
to which the pathways targeted and involved in infection are
the same, and whether these point to shared vulnerabilities
that could be useful therapeutically to quell infection by flavi-
viruses more generally.
The incidence of neurological symptoms associated with
Zika infection is a distinctive feature. While Guillain-Barre
syndrome, a weakness or paralysis caused by immune
attack on the peripheral nervous system, is frequently asso-
ciated with the aftermath of a variety of infections and so
its association with Zika is perhaps not altogether surprising,
the incidence of microcephaly in newborns is more myste-
rious. In contrast to Guillain-Barre, microcephaly impacts
the developing central nervous system and is likely due to
teratogenic effects of the virus, rather than the immune
response, and microcephaly has not previously been consid-
ered a potential repercussion of infection with flaviviruses.
That said, it is not entirely without precedent, as other types
of infection, such as toxoplasmosis and cytomegalovirus, are
also implicated in microcephaly. It should be cautioned that a
causal link between microcephaly and Zika virus has not yet
been firmly established (for a discussion of the growing evi-
dence read Vogel, 2016).
As concern grows, so does the impetus for developing a
Zika vaccine and for augmenting Aedes mosquito control
strategies. Here, recent efforts for dengue may point the
way forward. Brazil, which has been hit hard by both dengue
and Zika, recently approved for use the worlds first dengue
vaccine, Dengvaxia (from Sanofi Pasteur), which has also
been approved in Mexico and the Philippines. It is a chimeric
vaccine in which the envelope and pre-membrane genes of
the four serotypes of dengue replace those of an attenuated
strain of yellow fever. Other vaccine candidates, which
employ diverse approaches, are in phase I and II trials. No
such vaccines for Zika are at this stage of development,
and basic research directed at this goal will be key. Exploring
how neutralizing antibodies from previously infected individ-
uals block viral infection pinpoints potential targets for thera-
peutic development and provides insight into weak links in
Cell 164, February 11, 2016 2016 Elsevier Inc. 585
the viral life cycle. For instance, Jin et al. (2015) recently
defined a critical epitope bound by neutralizing monoclonal
antibodies for chikungunya that block both its entry and
release. Additionally, new vaccine design strategies also
offer promise, including the recent use of structure-based
engineering to create broadly-neutralizing antibodies for
dengue (Robinson et al., 2015). Targeting the mosquitos
that carry these diseases is another approach, bolstered by
the discovery that the wMel strain of Wolbachia, a genus of
bacteria that infect insects, blocks dengue transmission
(Walker et al., 2011). First tested in the lab, the wMel strain
of Wolbachia has since been introduced into natural mos-
quito populations with results showing promise (Hoffmann
et al., 2011; Nguyen et al., 2015)
The expansion of mosquito-borne flaviviruses over the past
twenty years, first dengue, then West Nile, and more recently
chikungunya and Zika have followed similar patterns (dis-
cussed by Fauci and Morens, 2016). This recurrence places
them at the forefront of research on emerging pathogens. In
light of this and the many open questions highlight the need
to accelerate research on their basic biology and vaccine
development, in order to position the research community
to rapidly respond to this and future epidemics.

REFERENCES

Enfissi, A., Codrington, J., Roosblad, J., Kazanji, M., and Rousset, D. (2016).
Lancet 387, 227.
Fauci, A.S., and Morens, D.M. (2016). N. Engl. J. Med. Published online
January 13, 2016. http://dx.doi.org/10.1056/NEJMp1600297.
Freire, C. C.M., Iamarino, A., Lima-Neto, D.F., Sall, A. A., and Zanotto, P. M. A.
(2015). BioRxiv http://dx.doi.org/10.1101/032839.
Hamel, R., Dejarnac, O., Wichit, S., Ekchariyawat, P., Neyret, A., Luplertlop, N.,
Perera-Lecoin, M., Surasombatpattana, P., Talignani, L., Thomas, F., et al.
(2015). J. Virol. 89, 88808896.
Hoffmann, A.A., Montgomery, B.L., Popovici, J., Iturbe-Ormaetxe, I.,
Johnson, P.H., Muzzi, F., Greenfield, M., Durkan, M., Leong, Y.S., Dong, Y.,
et al. (2011). Nature 476, 454457.
Jin, J., Liss, N.M., Chen, D.H., Liao, M., Fox, J.M., Shimak, R.M., Fong, R.H.,
Chafets, D., Bakkour, S., Keating, S., et al. (2015). Cell Rep. 13, 25532564.
Nguyen, T.H., Nguyen, H.L., Nguyen, T.Y., Vu, S.N., Tran, N.D., Le, T.N.,
Vien, Q.M., Bui, T.C., Le, H.T., Kutcher, S., et al. (2015). Parasit. Vectors 8, 563.
Robinson, L.N., Tharakaraman, K., Rowley, K.J., Costa, V.V., Chan, K.R.,
Wong, Y.H., Ong, L.C., Tan, H.C., Koch, T., Cain, D., et al. (2015). Cell 162,
493504.
Vogel, G. (2016). Science 351, 110111.
Walker, T., Johnson, P.H., Moreira, L.A., Iturbe-Ormaetxe, I., Frentiu, F.D.,
McMeniman, C.J., Leong, Y.S., Dong, Y., Axford, J., Kriesner, P., et al.
(2011). Nature 476, 450453.

Robert P. Kruger

Cell 164, February 11, 2016 2016 Elsevier Inc. 587