scholarly journals Human Parainfluenza Virus Type 1 C Proteins Are Nonessential Proteins That Inhibit the Host Interferon and Apoptotic Responses and Are Required for Efficient Replication in Nonhuman Primates

2008 ◽  
Vol 82 (18) ◽  
pp. 8965-8977 ◽  
Author(s):  
Emmalene J. Bartlett ◽  
Ann-Marie Cruz ◽  
Janice Esker ◽  
Adam Castaño ◽  
Henrick Schomacker ◽  
...  
Keyword(s):  
Virus Type ◽  
Parainfluenza Virus ◽  
Airway Epithelial ◽  
Efficient Replication ◽  
C Proteins ◽  
Induced Apoptosis ◽  
Human Parainfluenza Virus

ABSTRACT Recombinant human parainfluenza virus type 1 (rHPIV1) was modified to create rHPIV1-P(C−), a virus in which expression of the C proteins (C′, C, Y1, and Y2) was silenced without affecting the amino acid sequence of the P protein. Infectious rHPIV1-P(C−) was readily recovered from cDNA, indicating that the four C proteins were not essential for virus replication. Early during infection in vitro, rHPIV1-P(C−) replicated as efficiently as wild-type (wt) HPIV1, but its titer subsequently decreased coincident with the onset of an extensive cytopathic effect not observed with wt rHPIV1. rHPIV1-P(C−) infection, but not wt rHPIV1 infection, induced caspase 3 activation and nuclear fragmentation in LLC-MK2 cells, identifying the HPIV1 C proteins as inhibitors of apoptosis. In contrast to wt rHPIV1, rHPIV1-P(C−) and rHPIV1-CF170S, a mutant encoding an F170S substitution in C, induced interferon (IFN) and did not inhibit IFN signaling in vitro. However, only rHPIV1-P(C−) induced apoptosis. Thus, the anti-IFN and antiapoptosis activities of HPIV1 were separable: both activities are disabled in rHPIV1-P(C−), whereas only the anti-IFN activity is disabled in rHPIV1-CF170S. In African green monkeys (AGMs), rHPIV1-P(C−) was considerably more attenuated than rHPIV1-CF170S, suggesting that disabling the anti-IFN and antiapoptotic activities of HPIV1 had additive effects on attenuation in vivo. Although rHPIV1-P(C−) protected against challenge with wt HPIV1, its highly restricted replication in AGMs and in primary human airway epithelial cell cultures suggests that it might be overattenuated for use as a vaccine. Thus, the C proteins of HPIV1 are nonessential but have anti-IFN and antiapoptosis activities required for virulence in primates.

scholarly journals Role of Interferon in the Replication of Human Parainfluenza Virus Type 1 Wild Type and Mutant Viruses in Human Ciliated Airway Epithelium

2008 ◽  
Vol 82 (16) ◽  
pp. 8059-8070 ◽  
Author(s):  
Emmalene J. Bartlett ◽  
Margaret Hennessey ◽  
Mario H. Skiadopoulos ◽  
Alexander C. Schmidt ◽  
Peter L. Collins ◽  
...  
Keyword(s):  
Virus Type ◽  
Airway Epithelium ◽  
Parainfluenza Virus ◽  
Progeny Virus ◽  
Wild Type ◽  
Vaccine Candidates ◽  
C Proteins ◽  
Human Parainfluenza Virus

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.

scholarly journals Parainfluenza Virus Type 1 Induces Epithelial IL-8 Production via p38-MAPK Signalling

2014 ◽  
Vol 2014 ◽  
pp. 1-12 ◽  
Author(s):  
Miguel Ángel Galván Morales ◽  
Carlos Cabello Gutiérrez ◽  
Fidencio Mejía Nepomuceno ◽  
Leticia Valle Peralta ◽  
Elba Valencia Maqueda ◽  
...  
Keyword(s):  
Viral Replication ◽  
Cell Lines ◽  
Virus Type ◽  
Molecular Mechanisms ◽  
Parainfluenza Virus ◽  
Kinase Activation ◽  
Mitogen Activated Protein ◽  
Human Parainfluenza Virus

Human parainfluenza virus type 1 (HPIV-1) is the most common cause of croup in infants. The aim of this study was to describe molecular mechanisms associated with IL-8 production during HPIV-1 infection and the role of viral replication in MAPK synthesis and activation. Anin vitromodel of HPIV-1 infection in the HEp-2 and A549 cell lines was used; a kinetic-based ELISA for IL-8 detection was also used, phosphorylation of the mitogen-activated protein kinases (MAPKs) was identified by Western blot analysis, and specific inhibitors for each kinase were used to identify which MAPK was involved. Inactivated viruses were used to assess whether viral replication is required for IL-8 production. Results revealed a gradual increase in IL-8 production at different selected times, when phosphorylation of MAPK was detected. The secretion of IL-8 in the two cell lines infected with the HPIV-1 is related to the phosphorylation of the MAPK as well as viral replication. Inhibition of p38 suppressed the secretion of IL-8 in the HEp-2 cells. No kinase activation was observed when viruses were inactivated.

scholarly journals The P gene of human parainfluenza virus type 1 encodes P and C proteins but not a cysteine-rich V protein.

1991 ◽  
Vol 65 (6) ◽  
pp. 3406-3410 ◽  
Author(s):  
Y Matsuoka ◽  
J Curran ◽  
T Pelet ◽  
D Kolakofsky ◽  
R Ray ◽  
...  
Keyword(s):  
Virus Type ◽  
Parainfluenza Virus ◽  
V Protein ◽  
P Gene ◽  
C Proteins ◽  
Human Parainfluenza Virus

scholarly journals Attenuated Human Parainfluenza Virus Type 1 (HPIV1) Expressing the Fusion Glycoprotein of Human Respiratory Syncytial Virus (RSV) as a Bivalent HPIV1/RSV Vaccine

2015 ◽  
Vol 89 (20) ◽  
pp. 10319-10332 ◽  
Author(s):  
Natalie Mackow ◽  
Emérito Amaro-Carambot ◽  
Bo Liang ◽  
Sonja Surman ◽  
Matthias Lingemann ◽  
...  
Keyword(s):  
Respiratory Syncytial Virus ◽  
Virus Type ◽  
Deletion Mutation ◽  
Parainfluenza Virus ◽  
Temperature Sensitive ◽  
Bivalent Vaccine ◽  
Syncytial Virus ◽  
Human Parainfluenza Virus

ABSTRACTLive attenuated recombinant human parainfluenza virus type 1 (rHPIV1) was investigated as a vector to express the respiratory syncytial virus (RSV) fusion (F) glycoprotein, to provide a bivalent vaccine against RSV and HPIV1. The RSV F gene was engineered to include HPIV1 transcription signals and inserted individually into three gene locations in each of the two attenuated rHPIV1 backbones. Each backbone contained a single previously described attenuating mutation that was stabilized against deattenuation, specifically, a non-temperature-sensitive deletion mutation involving 6 nucleotides in the overlapping P/C open reading frames (ORFs) (CΔ170) or a temperature-sensitive missense mutation in the L ORF (LY942A). The insertion sites in the genome were pre-N (F1), N-P (F2), or P-M (F3) and were identical for both backbones.In vitro, the presence of the F insert reduced the rate of virus replication, but the final titers were the same as the final titer of wild-type (wt) HPIV1. High levels of RSV F expression in cultured cells were observed with rHPIV1-CΔ170-F1, -F2, and -F3 and rHPIV1-LY942A-F1. In hamsters, the rHPIV1-CΔ170-F1, -F2, and -F3 vectors were moderately restricted in the nasal turbinates, highly restricted in lungs, and genetically stablein vivo. Among the CΔ170vectors, the F1 virus was the most immunogenic and protective against wt RSV challenge. The rHPIV1-LY942Avectors were highly restrictedin vivoand were not detectably immunogenic or protective, indicative of overattenuation. The CΔ170-F1 construct appears to be suitably attenuated and immunogenic for further development as a bivalent intranasal pediatric vaccine.IMPORTANCEThere are no vaccines for the pediatric respiratory pathogens RSV and HPIV. We are developing live attenuated RSV and HPIV vaccines for use in virus-naive infants. Live attenuated RSV strains in particular are difficult to develop due to their poor growth and physical instability, but these obstacles could be avoided by the use of a vaccine vector. We describe the development and preclinical evaluation of live attenuated rHPIV1 vectors expressing the RSV F protein. Two different attenuated rHPIV1 backbones were each engineered to express RSV F from three different gene positions. The rHPIV1-CΔ170-F1 vector, bearing an attenuating deletion mutation (CΔ170) in the P/C gene and expressing RSV F from the pre-N position, was attenuated, stable, and immunogenic against the RSV F protein and HPIV1 in the hamster model and provided substantial protection against RSV challenge. This study provides a candidate rHPIV1-RSV-F vaccine virus suitable for continued development as a bivalent vaccine against two major childhood pathogens.

scholarly journals Involvement of Actin Microfilaments in the Replication of Human Parainfluenza Virus Type 3

1998 ◽  
Vol 72 (4) ◽  
pp. 2655-2662 ◽  
Author(s):  
Sanhita Gupta ◽  
Bishnu P. De ◽  
Judith A. Drazba ◽  
Amiya K. Banerjee
Keyword(s):  
Virus Type ◽  
Parainfluenza Virus ◽  
Actin Microfilaments ◽  
Viral Mrnas ◽  
Polymeric Form ◽  
Genomic Rnas ◽  
Human Parainfluenza Virus ◽  
Type 3

ABSTRACT Several studies indicate that paramyxoviruses require a specific cellular factor(s) for transcription of their genomic RNAs. We previously reported that the cellular cytoskeletal protein actin, in its polymeric form, participates in the transcription of human parainfluenza virus type 3 (HPIV3) in vitro. In the present study, we investigated the role of the polymeric form of actin, i.e., the actin microfilaments of the cytoskeletal framework, in the reproduction of HPIV3 in vivo. Pulse-chase labeling analyses indicate that the viral nucleocapsid-associated proteins, NP and P, are present predominantly in the cytoskeletal framework during infection. By in situ hybridization, we found that viral mRNAs and genomic RNA were synthesized from the nucleocapsids that were bound to the cytoskeletal framework. Double immunofluorescent labeling and confocal microscopy of the cytoarchitecture revealed that the viral nucleocapsids are specifically localized on the actin microfilaments. Treatment of cells with the actin-depolymerizing agent, cytochalasin D, resulted in the inhibition of viral RNA synthesis and ribonucleoprotein accumulation. These results strongly suggest that actin microfilaments play an important role in the replication of HPIV3.

scholarly journals The C Proteins of Human Parainfluenza Virus Type 1 (HPIV1) Control the Transcription of a Broad Array of Cellular Genes That Would Otherwise Respond to HPIV1 Infection

2008 ◽  
Vol 83 (4) ◽  
pp. 1892-1910 ◽  
Author(s):  
Jim B. Boonyaratanakornkit ◽  
Emmalene J. Bartlett ◽  
Emerito Amaro-Carambot ◽  
Peter L. Collins ◽  
Brian R. Murphy ◽  
...  
Keyword(s):  
Gene Expression ◽  
Transcription Factor ◽  
Virus Type ◽  
Parainfluenza Virus ◽  
Phenotypic Differences ◽  
Cellular Genes ◽  
C Proteins ◽  
Human Parainfluenza Virus ◽  
Expression Kinetics

ABSTRACTHuman parainfluenza virus type 1 (HPIV1) is an important respiratory pathogen in children and the most common cause of viral croup. We performed a microarray-based analysis of gene expression kinetics to examine how wild-type (wt) HPIV1 infection altered gene expression in human respiratory epithelial cells and what role beta interferon played in this response. We similarly evaluated HPIV1-P(C−), a highly attenuated and apoptosis-inducing virus that does not express any of the four C proteins, and HPIV1-CF170S, a less attenuated mutant that contains a single point mutation in C and, like wt HPIV1, does not efficiently induce apoptosis, to examine the role of the C proteins in controlling host gene expression. We also used these data to investigate whether the phenotypic differences between the two C mutants could be explained at the transcriptional level. Mutation or deletion of the C proteins of HPIV1 permitted the activation of over 2,000 cellular genes that otherwise would be repressed by HPIV1 infection. Thus, the C proteins profoundly suppress the response of human respiratory cells to HPIV1 infection. Cellular pathways targeted by the HPIV1 C proteins were identified and their transcriptional control was analyzed using bioinformatics. Transcription factor binding sites for IRF and NF-κB were overrepresented in some of the C protein-targeted pathways, but other pathways were dominated by less-known factors, such as forkhead transcription factor FOXD1. Surprisingly, the host responses to the P(C−) and CF170Smutants were very similar, and only subtle differences in the expression kinetics of caspase 3 and TRAIL receptor 2 were observed. Thus, changes in host cell transcription did not reflect the striking phenotypic differences observed between these two viruses.

scholarly journals The C Proteins of Human Parainfluenza Virus Type 1 Block IFN Signaling by Binding and Retaining Stat1 in Perinuclear Aggregates at the Late Endosome

PLoS ONE ◽  
2012 ◽  
Vol 7 (2) ◽  
pp. e28382 ◽  
Author(s):  
Henrick Schomacker ◽  
Rebecca M. Hebner ◽  
Jim Boonyaratanakornkit ◽  
Sonja Surman ◽  
Emerito Amaro-Carambot ◽  
...  
Keyword(s):  
Virus Type ◽  
Late Endosome ◽  
Parainfluenza Virus ◽  
C Proteins ◽  
Human Parainfluenza Virus
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