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VIRAL INFECTION
Filamentous viruses prevail under pressure
Enveloped viruses often produce particles with varying shapes. Elongated particles are shown to be crucial for
maintaining infectivity under conditions in which viral glycoprotein activity is compromised.
Anice C. Lowen
M
any viruses produce particles
that are heterogeneous in length
and shape, ranging from spheres
to elongated filaments. Spherical particles
are sufficient to envelope the viral genome
and ferry it to new target cells or a new
host. Filaments can also fulfil this essential
function but, owing to their larger surface
area, use a much greater store of structural
resources to accomplish the same task.
Why, then, have viruses evolved to produce
filaments? Virologists have puzzled over
this question for decades. In this issue of
Nature Microbiology, Li et al. set out to
address this long-standing conundrum and
in doing so define the functional relevance
of particle length for virus attachment to,
and entry into, target cells1. They find that
when attachment and entry are constrained,
for example by host immune responses,
spherical particles often fail to complete the
task of genome delivery, whereas elongated
particles prevail. The production of particles
that vary in length may serve to enable
viral infection under diverse and changing
conditions within and between hosts.
In the 1940s, examination of influenza A
virus by electron microscopy revealed that
virus particles can take on both spherical
and filamentous morphologies, with a
broad range of particle lengths observed2
(Fig. 1). Upon passage in the laboratory, the
ability of an influenza isolate to produce
filamentous particles is often lost, suggesting
that filamentous forms offer a fitness
advantage in hosts but not in laboratory
cultures3. Owing to their increased surface
area, elongated particles incorporate a
greater number of haemagglutinin (HA)
proteins, the glycoprotein responsible for
viral attachment to and entry into target
cells4. Nevertheless, increased particle length
does not seem to increase infectivity, at least
under standard laboratory conditions5,6.
Seeking to uncover the role of
filamentous viral particles during infection,
Li et al. hypothesized that production
of particles of varied length ensures that
infection can proceed under a range of
environmental pressures. To test this idea,
they investigated whether particle shape
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affects the sensitivity of viruses to cell-entry
constraints, including antibody interference
with virus attachment, inhibition of
membrane fusion and incomplete
proteolytic activation of the fusion
glycoprotein. Influenza A virus was used as
the test case for this work and, to extend the
concepts to a second viral system, influenza
virus particles carrying the Ebola virus
glycoprotein (GP) were also examined.
To test the effect of particle length
on infectivity, the authors needed to
separate long and short particles. This was
challenging because of their similar densities
and identical genetic components, but
Li et al. overcame this problem by applying
serial centrifugation to enrich filaments
in the pellet and leave (mainly) spheres in 0.5 μm
the supernatant. The infectivity of these
two virus populations was then compared
and was found to be the same under Fig. 1 | Influenza A virus particles can be
baseline conditions. However, differences in spherical or filamentous, with a wide range of
infectivity were noted when two antibodies filament lengths produced. Reproduced with
that target HA were added. One of the permission from ref. 8, American Society for
antibodies solely blocks the attachment of Microbiology.
HA to cell surfaces, and another prevents
HA-mediated membrane fusion. In both
cases, filamentous virus populations were To investigate the mechanism underlying
less sensitive to neutralization. Thus, in the filament resistance to the fusion-inhibiting
context of cell-entry constraints imposed by antibody, single-particle membrane fusion
neutralizing antibodies, elongated particles experiments were performed. Spherical or
were more infectious. filamentous virus particles were overlaid
To test whether the attachment of the onto a lipid bilayer, and fusion of the viral
virion to the cell surface was altered by the envelope with that lipid bilayer was triggered
application of an antibody, the adherence by reducing the pH. At low pH, HA on the
of virus particles to cell surfaces was virus particles inserts a fusion peptide into
assayed. In this experiment, cells were the opposing membrane before pulling back,
chilled on ice (so that particles could causing the two membranes to fuse. This
attach but could not be taken up), and process usually initiates infection and allows
viral genomes at the cell surface were delivery of the viral genome to the cell.
detected using in situ hybridization. This Li et al. found that both virus populations
method was used to enumerate particles initiated a similar number of fusion events,
because genome copies per virus do not but elongated particles mediated fusion
vary with particle shape. Consistent with more quickly. Filamentous viruses were
the infectivity results, virus populations also found to be relatively resistant to
enriched in filaments or spheres were the fusion-blocking antibody, as shown
found to attach with equal efficiency under by monitoring the proportion of single
baseline conditions, but more filaments particles initiating fusion in the presence
were able to bind to the cell surface of an antibody. So, whilst spherical and
when viruses were pretreated with an filamentous particles are equally competent
attachment-blocking antibody. for membrane fusion, under the constraint
Nature Microbiology | VOL 6 | May 2021 | 536–537 | www.nature.com/naturemicrobiology

imposed by a fusion-blocking antibody,
filamentous particles mediate fusion more
efficiently.
The authors propose a model in which
the elongated surface of a filament allows
a larger number of HA trimers to interface
with the target membrane and thereby
increases the number of potential fusion
clusters per virion. A fusion cluster is a
group of neighbouring HAs that act together
to precipitate fusion. In this model, longer
particles allow more fusion clusters to
form, such that the first productive fusion
event occurs faster. In addition, when a
subset of HA molecules is bound by an
antibody, filaments would have an increased
likelihood of fusion clusters forming at all.
The experimental results are qualitatively
consistent with both of these concepts. To
further solidify their conclusions, Li et al.
used a simulation model of fusion dynamics
and showed that the measured trends of
time to fusion and yield of fusion events are
attributable to differences in the surface area
of the virus interacting with the membrane.
To extend their investigation to an
additional pleomorphic virus, Li et al.
used a model system in which influenza
virus particles are decorated with Ebola
virus GP rather than HA. Here, delivery of
the viral genome to cells is dependent on
attachment and fusion mediated by GP. In
this system, populations of elongated virus
particles showed more efficient infection
both in the absence and the presence of
an antibody that blocks GP-mediated
fusion. While the similar outcomes seen
with HA- and GP-decorated viruses
suggest that the functional importance
of filamentous particles is generalizable
across diverse virus families, examination
of Ebola virus particles is needed to solidify
this conclusion. In the system used, GP
distribution on particles and the particle
Nature Microbiology | VOL 6 | May 2021 | 536–537 | www.nature.com/naturemicrobiology
lengths represented likely differ from Ebola
virus itself.
Whilst Li et al. focused their investigation
on attachment and fusion, virus particle
shape may affect additional steps of the viral
life cycle. The efficiency with which nascent
viral particles are formed, released from
producer cells and dispersed within and
between hosts are all areas where particle
shape could be highly significant. The
influenza virus neuraminidase (NA) protein
facilitates release and dispersal and, together
with HA, is the second major glycoprotein
at the particle surface. Further investigation
centred on the implications of particle shape
for NA function is needed and, since HA
and NA have opposing activities, the effect
of particle shape on HA and NA balance is
highly relevant.
The authors propose an appealing model
in which pleomorphic particle shape acts
as a bet-hedging strategy7. In bet-hedging,
organisms invest in multiple strategies at
once, potentially reducing overall fitness
in their typical environment but ensuring
increased fitness under stress. Spherical
influenza viruses may be the dominant
source of infection in naïve hosts as their
uptake and production are more efficient.
These smaller particles may also be critical
for transmission mediated by small droplet
aerosols. However, in the presence of
HA-specific humoral immunity, mucus
— or other factors interfering with viral
entry — filaments may be needed to sustain
infection. The seemingly wasteful elongated
particles may therefore become essential as
their spherical counterparts are efficiently
neutralized. Extending this line of reasoning,
pleomorphic particles might conceivably
enable emergence of non-endemic viruses
into a new host species. It is possible that
filamentous particles may compensate for
cell-entry deficiencies in a new host, thereby
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enabling a low level of propagation and,
in turn, the opportunity for adaptation
by mutation. These concepts are likely to
extend to diverse viruses with pleomorphic
particle shape, providing a clear
evolutionary rationale for a long-observed
biological phenomenon.
More broadly, the findings of Li et al.
raise questions about how the shape of virus
particles relates to infectivity. Many viruses
produce particles of uniform shape and
size, perhaps owing to constraints imposed
at the particle assembly stage. Virion
uniformity would preclude bet-hedging
based on varied particle size, so the
question arises as to whether these viruses
have evolved different means of ensuring
infectivity in variable environments. ❐
Anice C. Lowen    ✉
Department of Microbiology and Immunology,
Emory University School of Medicine,
Atlanta, GA, USA.
✉e-mail: anice.lowen@emory.edu
Published online: 29 April 2021
https://doi.org/10.1038/s41564-021-00903-1
References
1. Li, T. et al. Nat. Microbiol. https://doi.org/10.1038/s41564-021-
00877-0 (2021).
2. Mosley, V. M. & Wyckoff, R. W. G. Nature 157, 263
(1946).
3. Choppin, P. W., Murphy, J. S. & Tamm, I. J. Exp. Med. 112,
945–952 (1960).
4. Roberts, P. C., Lamb, R. A. & Compans, R. W. Virology 240,
127–137 (1998).
5. Seladi-Schulman, J., Campbell, P. J., Suppiah, S., Steel, J. &
Lowen, A. C. PLoS ONE 9, e112462 (2014).
6. Dadonaite, B., Vijayakrishnan, S., Fodor, E., Bhella, D. &
Hutchinson, E. C. J. Gen. Virol. 97, 1755–1764 (2016).
7. Slatkin, M. Nature 250, 704–705 (1974).
8. Seladi-Schulman, J., Steel, J. & Lowen, A. C. J. Virol. 24,
13343–13353 (2013).
Competing interests
The author declares no competing interests.
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