Attenuating mutations in the P/C gene of human parainfluenza virus type 1 (HPIV1) vaccine candidates abrogate the inhibition of both induction and signaling of type I interferon (IFN) by wild-type HPIV1

ABSTRACT Human parainfluenza virus type 1 (HPIV1) is a significant cause of pediatric respiratory disease in the upper and lower airways. An in vitro model of human ciliated airway epithelium (HAE), a useful tool for studying respiratory virus-host interactions, was used in this study to show that HPIV1 selectively infects ciliated cells within the HAE and that progeny virus is released from the apical surface with little apparent gross cytopathology. In HAE, type I interferon (IFN) is induced following infection with an HPIV1 mutant expressing defective C proteins with an F170S amino acid substitution, rHPIV1-CF170S, but not following infection with wild-type HPIV1. IFN induction coincided with a 100- to 1,000-fold reduction in virus titer, supporting the hypothesis that the HPIV1 C proteins are critical for the inhibition of the innate immune response. Two recently characterized live attenuated HPIV1 vaccine candidates expressing mutant C proteins were also evaluated in HAE. The vaccine candidates, rHPIV1-CR84G/Δ170HNT553ALY942A and rHPIV1-CR84G/Δ170HNT553ALΔ1710-11, which contain temperature-sensitive (ts) attenuating (att) and non-ts att mutations, were highly restricted in growth in HAE at permissive (32°C) and restrictive (37°C) temperatures. The viruses grew slightly better at 37°C than at 32°C, and rHPIV1-CR84G/Δ170HNT553ALY942A was less attenuated than rHPIV1-CR84G/Δ170HNT553ALΔ1710-11. The level of replication in HAE correlated with that previously observed for African green monkeys, suggesting that the HAE model has potential as a tool for the preclinical evaluation of HPIV1 vaccines, although how these in vitro data will correlate with vaccine virus replication in seronegative human subjects remains to be seen.

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Human parainfluenza virus type 1 regulates cholesterol biosynthesis and establishes quiescent infection in human airway cells

PLoS Pathogens ◽

10.1371/journal.ppat.1009908 ◽

2021 ◽

Vol 17 (9) ◽

pp. e1009908

Author(s):

Yuki Kurebayashi ◽

Shringkhala Bajimaya ◽

Masahiro Watanabe ◽

Nicholas Lim ◽

Michael Lutz ◽

...

Keyword(s):

Virus Type ◽

Cholesterol Biosynthesis ◽

Parainfluenza Virus ◽

Type I ◽

Human Airway ◽

Virion Release ◽

Infected Cells ◽

Human Parainfluenza Virus ◽

Airway Cells

Human parainfluenza virus type 1 (hPIV1) and 3 (hPIV3) cause seasonal epidemics, but little is known about their interaction with human airway cells. In this study, we determined cytopathology, replication, and progeny virion release from human airway cells during long-term infection in vitro. Both viruses readily established persistent infection without causing significant cytopathic effects. However, assembly and release of hPIV1 rapidly declined in sharp contrast to hPIV3 due to impaired viral ribonucleocapsid (vRNP) trafficking and virus assembly. Transcriptomic analysis revealed that both viruses induced similar levels of type I and III IFNs. However, hPIV1 induced specific ISGs stronger than hPIV3, such as MX2, which bound to hPIV1 vRNPs in infected cells. In addition, hPIV1 but not hPIV3 suppressed genes involved in lipid biogenesis and hPIV1 infection resulted in ubiquitination and degradation of 3-hydroxy-3-methylglutaryl-coenzyme A reductase, a rate limiting enzyme in cholesterol biosynthesis. Consequently, formation of cholesterol-rich lipid rafts was impaired in hPIV1 infected cells. These results indicate that hPIV1 is capable of regulating cholesterol biogenesis, which likely together with ISGs contributes to establishment of a quiescent infection.

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A novel human parainfluenza virus type 1 (HPIV1) with separated P and C genes is useful for generating C gene mutants for evaluation as live-attenuated virus vaccine candidates

Vaccine ◽

10.1016/j.vaccine.2009.10.069 ◽

2010 ◽

Vol 28 (3) ◽

pp. 767-779 ◽

Cited By ~ 6

Author(s):

Emmalene J. Bartlett ◽

Ann-Marie Cruz ◽

Jim Boonyaratanakornkit ◽

Janice Esker ◽

Adam Castaño ◽

...

Keyword(s):

Virus Type ◽

Virus Vaccine ◽

Parainfluenza Virus ◽

Vaccine Candidates ◽

Attenuated Virus ◽

Human Parainfluenza Virus

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Attenuation and efficacy of human parainfluenza virus type 1 (HPIV1) vaccine candidates containing stabilized mutations in the P/C and L genes

Virology Journal ◽

10.1186/1743-422x-4-67 ◽

2007 ◽

Vol 4 (1) ◽

pp. 67 ◽

Cited By ~ 18

Author(s):

Emmalene J Bartlett ◽

Adam Castaño ◽

Sonja R Surman ◽

Peter L Collins ◽

Mario H Skiadopoulos ◽

...

Keyword(s):

Virus Type ◽

Parainfluenza Virus ◽

Vaccine Candidates ◽

Human Parainfluenza Virus

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Generation of Recombinant Human Parainfluenza Virus Type 1 Vaccine Candidates by Importation of Temperature-Sensitive and Attenuating Mutations from Heterologous Paramyxoviruses

Journal of Virology ◽

10.1128/jvi.78.4.2017-2028.2004 ◽

2004 ◽

Vol 78 (4) ◽

pp. 2017-2028 ◽

Cited By ~ 32

Author(s):

Jason T. Newman ◽

Jeffrey M. Riggs ◽

Sonja R. Surman ◽

Josephine M. McAuliffe ◽

Teresa A. Mulaikal ◽

...

Keyword(s):

Respiratory Tract ◽

Virus Type ◽

Neutralizing Antibodies ◽

Effective Means ◽

Parainfluenza Virus ◽

Temperature Sensitive ◽

C Protein ◽

Vaccine Candidates ◽

Human Parainfluenza Virus

ABSTRACT Human parainfluenza virus type 1 (HPIV1) is a significant cause of respiratory tract disease in infants and young children for which a vaccine is needed. In the present study, we sought to attenuate HPIV1 by the importation of one or more known attenuating point mutations from heterologous paramyxoviruses into homologous sites in HPIV1. The introduced mutations were derived from three attenuated paramyxoviruses: (i) HPIV3cp45, a live-attenuated HPIV3 vaccine candidate containing multiple attenuating mutations; (ii) the respiratory syncytial virus cpts530 with an attenuating mutation in the L polymerase protein; and (iii) a murine PIV1 (MPIV1) attenuated by a mutation in the accessory C protein. Recombinant HPIV1 (rHPIV1) mutants bearing a single imported mutation in C, any of three different mutations in L, or a pair of mutations in F exhibited a 100-fold or greater reduction in replication in the upper or lower respiratory tract of hamsters. Both temperature-sensitive (ts) (mutations in the L and F proteins) and non-ts (the mutation in the C protein) attenuating mutations were identified. rHPIV1 mutants containing a combination of mutations in L were generated that were more attenuated than viruses bearing the individual mutations, showing that the systematic accretion of mutations can yield progressive increases in attenuation. Hamsters immunized with rHPIV1 mutants bearing one or two mutations developed neutralizing antibodies and were resistant to challenge with wild-type HPIV1. Thus, importation of attenuating mutations from heterologous viruses is an effective means for rapidly identifying mutations that attenuate HPIV1 and for generating live-attenuated HPIV1 vaccine candidates.

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The Long Noncoding Region of the Human Parainfluenza Virus Type 1 F Gene Contributes to the Read-Through Transcription at the M-F Gene Junction

Journal of Virology ◽

10.1128/jvi.76.16.8244-8251.2002 ◽

2002 ◽

Vol 76 (16) ◽

pp. 8244-8251 ◽

Cited By ~ 31

Author(s):

Tatiana Bousse ◽

Tatyana Matrosovich ◽

Allen Portner ◽

Atsushi Kato ◽

Yoshiyuki Nagai ◽

...

Keyword(s):

Virus Type ◽

Parainfluenza Virus ◽

Noncoding Sequence ◽

Wild Type ◽

F Gene ◽

Transcriptional Termination ◽

Infected Cells ◽

Human Parainfluenza Virus ◽

Gene Junction

ABSTRACT Sendai virus (SV) and human parainfluenza virus type 1 (hPIV1) have genomes consisting of nonsegmented negative-sense RNA in which the six genes are separated by well-conserved intergenic (IG) sequences and transcriptional start (S) and end signals. In hPIV1-infected cells, transcriptional termination at the M-F gene junction is ineffective; a large number of M-F read-through transcripts are produced (T. Bousse, T. Takimoto, K. G. Murti, and A. Portner, Virology 232:44-52, 1997). In contrast, few M-F read-through transcripts are detected in SV-infected cells. Sequence analysis indicated that the hPIV1 IG and S sequences in the M-F junction differ from those of SV. Furthermore, the hPIV1 F gene contains an unusually long noncoding sequence. To identify the cis-acting elements that prevent transcriptional termination at the M-F junction, we rescued recombinant SV (rSVhMFjCG) in which its M-F gene junction was replaced by that of hPIV1. Cells infected with rSVhMFjCG produced an abundance of M-F read-through transcripts; this result indicated that the hPIV1 M-F junction is responsible for inefficient termination. When one or both of the IG and S sites in rSVhMFjCG were replaced by those of SV, the efficiency of transcriptional termination increased but not to the level observed in wild-type SV-infected cells. Deletion of most of the long noncoding region of the hPIV1 F gene in rSVhMFjCG in addition to the mutations in IG and S signals resulted in efficient termination that was equivalent to the level observed in wild-type virus-infected cells. Therefore, the long noncoding sequence of the hPIV1 F gene contains cis-acting element(s) that affects transcriptional termination. Our evaluation of the effect of inefficient transcriptional termination on viral replication in culture revealed that cells infected with rSVhMFjCG produced less F protein than cells infected with wild-type SV and that assembly of the recombinant SV in culture was less efficient. These phenotypes seem to be responsible for the extended survival of mice infected with rSVhMFjCG.

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Human parainfluenza virus type I (HPIV1) vaccine candidates designed by reverse genetics are attenuated and efficacious in African green monkeys

Vaccine ◽

10.1016/j.vaccine.2005.04.035 ◽

2005 ◽

Vol 23 (38) ◽

pp. 4631-4646 ◽

Cited By ~ 34

Author(s):

Emmalene J. Bartlett ◽

Emerito Amaro-Carambot ◽

Sonja R. Surman ◽

Jason T. Newman ◽

Peter L. Collins ◽

...

Keyword(s):

Virus Type ◽

Reverse Genetics ◽

Parainfluenza Virus ◽

Type I ◽

Vaccine Candidates ◽

Human Parainfluenza Virus ◽

Green Monkeys

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Introducing point and deletion mutations into the P/C gene of human parainfluenza virus type 1 (HPIV1) by reverse genetics generates attenuated and efficacious vaccine candidates

Vaccine ◽

10.1016/j.vaccine.2005.10.047 ◽

2006 ◽

Vol 24 (14) ◽

pp. 2674-2684 ◽

Cited By ~ 20

Author(s):

Emmalene J. Bartlett ◽

Emerito Amaro-Carambot ◽

Sonja R. Surman ◽

Peter L. Collins ◽

Brian R. Murphy ◽

...

Keyword(s):

Virus Type ◽

Reverse Genetics ◽

Parainfluenza Virus ◽

Vaccine Candidates ◽

Deletion Mutations ◽

Efficacious Vaccine ◽

Human Parainfluenza Virus

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The human parainfluenza virus type-1 prototypic strain contains a heat-labile hemagglutinin-neuraminidase protein

Virus Research ◽

10.1016/0168-1702(94)90063-9 ◽

1994 ◽

Vol 32 (1) ◽

pp. 85-92 ◽

Cited By ~ 2

Author(s):

Toru Takimoto ◽

Allen Portner

Keyword(s):

Virus Type ◽

Parainfluenza Virus ◽

Heat Labile ◽

Human Parainfluenza Virus

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Mechanism of Interference Mediated by Human Parainfluenza Virus Type 3 Infection

Journal of Virology ◽

10.1128/jvi.74.24.11792-11799.2000 ◽

2000 ◽

Vol 74 (24) ◽

pp. 11792-11799 ◽

Cited By ~ 21

Author(s):

Maria-Arantxa Horga ◽

G. Luca Gusella ◽

Olga Greengard ◽

Natalia Poltoratskaia ◽

Matteo Porotto ◽

...

Keyword(s):

Virus Type ◽

Fluorescent Protein ◽

Parainfluenza Virus ◽

Wild Type ◽

Challenge Virus ◽

Neuraminidase Activity ◽

Definitive Evidence ◽

Viral Interference ◽

Human Parainfluenza Virus ◽

Type 3

ABSTRACT Viral interference is characterized by the resistance of infected cells to infection by a challenge virus. Mechanisms of viral interference have not been characterized for human parainfluenza virus type 3 (HPF3), and the possible role of the neuraminidase (receptor-destroying) enzyme of the hemagglutinin-neuraminidase (HN) glycoprotein has not been assessed. To determine whether continual HN expression results in depletion of the viral receptors and thus prevents entry and cell fusion, we tested whether cells expressing wild-type HPF3 HN are resistant to viral infection. Stable expression of wild-type HN-green fluorescent protein (GFP) on cell membranes in different amounts allowed us to establish a correlation between the level of HN expression, the level of neuraminidase activity, and the level of protection from HPF3 infection. Cells with the highest levels of HN expression and neuraminidase activity on the cell surface were most resistant to infection by HPF3. To determine whether this resistance is attributable to the viral neuraminidase, we used a cloned variant HPF3 HN that has two amino acid alterations in HN leading to the loss of detectable neuraminidase activity. Cells expressing the neuraminidase-deficient variant HN-GFP were not protected from infection, despite expressing HN on their surface at levels even higher than the wild-type cell clones. Our results demonstrate that the HPF3 HN-mediated interference effect can be attributed to the presence of an active neuraminidase enzyme activity and provide the first definitive evidence that the mechanism for attachment interference by a paramyxovirus is attributable to the viral neuraminidase.

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