JOURNAL OF VIROLOGY, Mar. 2006, p. 2589–2595
Copyright © 2006, American Society for Microbiology. All Rights Reserved.
Vol. 80, No. 6
Age-Dependent Poliovirus Replication in the Mouse Central Nervous
System Is Determined by Internal Ribosome
Entry Site-Mediated Translation
Steven Kauder,† Sherry Kan,‡ and Vincent R. Racaniello*
Department of Microbiology, Columbia University College of Physicians and Surgeons, New York, New York 10032
Received 2 June 2005/Accepted 21 December 2005
Mouse cells are not permissive for the replication of human rhinovirus type 2 (HRV2). To determine the role
of the HRV2 internal ribosome entry site (IRES) in determining species specificity, a recombinant poliovirus
(P1/HRV2) was constructed by substituting the poliovirus IRES with the IRES from HRV2. This recombinant
virus replicated in all human and murine cell lines examined, demonstrating that the HRV2 IRES does not
limit viral replication in transformed murine cells. P1/HRV2 replicated in the brain and spinal cord in
neonatal but not adult mice transgenic for the poliovirus receptor, CD155. Passage of P1/HRV2 in mice led to
selection of a virus that caused paralysis in neonatal mice. To determine the relationship between HRV2
IRES-mediated translation and replication of P1/HRV2 in mice, recombinant human adenoviruses were used
to express bicistronic mRNAs in murine organs. The results demonstrate that the HRV2 IRES mediates
translation in organs of neonatal but not adult mice. These findings show that HRV2 IRES-mediated trans-
lation is a determinant of virus replication in the murine brain and spinal cord and suggest that the IRES
determines the species specificity of HRV2 infection.
Human rhinoviruses (HRVs) are nonenveloped, positive-
stranded RNA viruses of the family Picornaviridae (8). Approx-
imately 100 HRV serotypes have been identified and divided
into two groups based on receptor usage. The receptor for
major group HRVs is human intracellular adhesion molecule 1
(hICAM-1), and the receptor for minor group HRVs is human
low-density lipoprotein receptor (8). HRVs replicate in the
epithelium of the human respiratory tract (1, 2, 11, 41) and are
responsible for the majority of common cold infections of
humans (36). Only humans develop clinical symptoms after
HRV infection; experimental asymptomatic infections have
been documented in chimpanzees (10) and gibbons (35). A
rodent model for infection with wild-type HRV has not been
A small-animal model of HRV infection would be useful for
elucidating the mechanisms of HRV-induced pathogenesis and
to develop therapeutic interventions. The limited host range of
most HRV serotypes (8) has hindered establishment of such a
model. Mouse cells are not permissive for the replication of
minor group serotypes, despite the ability of these viruses to
enter these cells after binding the murine homolog of human
low-density lipoprotein receptor. Two exceptions are HRV1A
(37) and a variant of HRV2 (HRV2/L) selected by passage in
mouse cells (43). The changes required for growth of HRV2/L
in mouse cells have not been identified (43). HRV2/L produces
altered P2 proteins in infected cells, suggesting that the block
to HRV2 replication in murine cells could be due to a defect
in RNA replication (29). This conclusion is supported by the
finding that HRV2/L is less sensitive than HRV2 to chemical
inhibitors of RNA replication (43). It is believed that mouse
cell lines are neither permissive nor susceptible to infection
with most major group HRVs. While the block to replication
of HRV14 and HRV16 in mouse L cells is relieved upon
synthesis of hICAM-1, HRV39 fails to replicate (17). Passage
of HRV39 in mouse cells producing hICAM-1 led to the iden-
tification of a virus, HRV39/L, that can replicate in these cells
(16). Amino acid changes in viral proteins 2B and 3A mediate
HRV39 growth in mouse cells.
HRV host range may be determined in part by translation of
the viral mRNA. The 5? ends of picornaviral mRNAs are not
linked to a 7-methylguanosine cap structure and cannot be
translated by 5? end-dependent initiation as are most cellular
mRNAs (6, 24, 25). Instead, initiation on picornaviral RNA is
mediated by the viral internal ribosome entry site (IRES), a
cis-acting RNA sequence that binds ribosomes in the absence
of an mRNA cap or free 5? end (3, 4, 7, 32, 33). It has been
suggested that the limited tropism of poliovirus, a member of
the same virus family as HRV, is determined by organ-specific
differences in IRES-mediated translation (5, 14, 42). This hy-
pothesis has been disproved by the finding that the IRES of
poliovirus and other picornaviruses mediates translation in
organs that are not permissive for virus replication (9, 20, 39).
Results obtained with a transgenic mouse model for polio-
myelitis have implicated the HRV2 IRES as a determinant of
HRV2 host range (14, 15). Poliovirus infection of mice trans-
genic for the human poliovirus receptor gene, CD155 (TgPVR
mice) leads to virus replication in neurons of the brain and
spinal cord and flaccid paralysis (23, 38). Unlike wild-type
poliovirus, a recombinant poliovirus strain with the IRES of
HRV2 does not replicate or cause disease in CD155 transgenic
* Corresponding author. Mailing address: Department of Microbi-
ology, Columbia University College of Physicians and Surgeons, 701
W. 168th St., New York, NY 10032. Phone: (212) 305-5707. Fax: (212)
305-5106. E-mail: email@example.com.
† Present address: Gladstone Institute of Virology and Immunology,
University of California, San Francisco, CA 94141-9100.
‡ Present address: The Rockefeller University, New York, NY
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VOL. 80, 2006 RHINOVIRUS IRES AND HOST RANGE2595