scholarly journals A Novel Chimeric Adenoassociated Virus 2/Human Bocavirus 1 Parvovirus Vector Efficiently Transduces Human Airway Epithelia

2013 ◽  
Vol 21 (12) ◽  
pp. 2181-2194 ◽  
Author(s):  
Ziying Yan ◽  
Nicholas W Keiser ◽  
Yi Song ◽  
Xuefeng Deng ◽  
Fang Cheng ◽  
...  
Keyword(s):  
Human Bocavirus ◽  
Airway Epithelia ◽  
Human Airway ◽  
Adenoassociated Virus ◽  
Human Airway Epithelia

Hairpin transfer-independent Parvovirus DNA Replication Produces Infectious Virus

2021 ◽  
Author(s):  
Weiran Shen ◽  
Zekun Wang ◽  
Kang Ning ◽  
Fang Cheng ◽  
John F. Engelhardt ◽  
...  
Keyword(s):  
Dna Replication ◽  
Hek293 Cells ◽  
Progeny Virus ◽  
Human Bocavirus ◽  
Viral Dna ◽  
Airway Epithelia ◽  
Viral Dna Replication ◽  
Human Airway ◽  
The Right ◽  
Human Airway Epithelia

Parvoviruses package a linear single-stranded DNA genome with hairpin structures at both ends. It has been thought that terminal hairpin sequences are indispensable for viral DNA replication. Here, we provide evidence that the hairpin-deleted duplex genomes of human bocavirus 1 (HBoV1) replicate in human embryonic kidney (HEK) 293 cells. We propose an alternative model for HBoV1 DNA replication in which the leading strand can initiate strand-displacement without “hairpin-transfer.” The transfection of the HBoV1 duplex genomes that retain a minimal replication origin at the right-end ( OriR ), but with extensive deletions in the right-end hairpin (REH), generated viruses in HEK293 cells at a level 10-20 times lower than the wild-type (WT) duplex genome. Importantly, these viruses that have a genome with various deletions after the OriR , but not the one retaining only the OriR , replicated in polarized human airway epithelia. We discovered that the 18-nt sequence (nt 5,403-5,420) beyond the OriR was sufficient to confer virus replication in polarized human airway epithelia, although its progeny virus production was ∼5 times lower than that of the WT virus. Thus, our study demonstrates that hairpin transfer-independent productive parvovirus DNA replication can occur. Importance Hairpin transfer-independent parvovirus replication was modeled with human bocavirus 1 (HBoV1) duplex genomes whose 5’ hairpin structure was ablated by various deletions. In HEK293 cells, these duplex viral genomes with ablated 5’/hairpin sequence replicated efficiently and generated viruses that productively infected polarized human airway epithelium. Thus, for the first time, we reveal a previously unknown phenomenon that the productive parvovirus DNA replication does not depend on the hairpin sequence at REH to initiate “rolling hairpin” DNA replication. Notably, the intermediates of viral DNA replication, as revealed two-dimensional electrophoresis, from transfections of hairpin sequence-deleted duplex genome and full-length genome in HEK293 cells, as well as from virus infection of polarized human airway epithelia are similar. Thus, the establishment of the hairpin transfer-independent parvoviral DNA replication deepens our understanding in viral DNA replication and may have implications in development of parvovirus-based viral vectors with alternative properties.

scholarly journals Human Parvovirus Infection of Human Airway Epithelia Induces Pyroptotic Cell Death by Inhibiting Apoptosis

2017 ◽  
Vol 91 (24) ◽  
Author(s):  
Xuefeng Deng ◽  
Wei Zou ◽  
Min Xiong ◽  
Zekun Wang ◽  
John F. Engelhardt ◽  
...  
Keyword(s):  
Cell Death ◽  
Epithelial Cells ◽  
Airway Epithelial Cells ◽  
Human Bocavirus ◽  
Airway Epithelial ◽  
Airway Epithelia ◽  
Human Airway ◽  
Caspase 1 ◽  
Tract Infections ◽  
Human Airway Epithelia

ABSTRACT Human bocavirus 1 (HBoV1) is a human parvovirus that causes acute respiratory tract infections in young children. In this study, we confirmed that, when polarized/well-differentiated human airway epithelia are infected with HBoV1 in vitro, they develop damage characterized by barrier function disruption and cell hypotrophy. Cell death mechanism analyses indicated that the infection induced pyroptotic cell death characterized by caspase-1 activation. Unlike infections with other parvoviruses, HBoV1 infection did not activate the apoptotic or necroptotic cell death pathway. When the NLRP3-ASC-caspase-1 inflammasome-induced pathway was inhibited by short hairpin RNA (shRNA), HBoV1-induced cell death dropped significantly; thus, NLRP3 mediated by ASC appears to be the pattern recognition receptor driving HBoV1 infection-induced pyroptosis. HBoV1 infection induced steady increases in the expression of interleukin 1α (IL-1α) and IL-18. HBoV1 infection was also associated with the marked expression of the antiapoptotic genes BIRC5 and IFI6. When the expression of BIRC5 and/or IFI6 was inhibited by shRNA, the infected cells underwent apoptosis rather than pyroptosis, as indicated by increased cleaved caspase-3 levels and the absence of caspase-1. BIRC5 and/or IFI6 gene inhibition also significantly reduced HBoV1 replication. Thus, HBoV1 infection of human airway epithelial cells activates antiapoptotic proteins that suppress apoptosis and promote pyroptosis. This response may have evolved to confer a replicative advantage, thus allowing HBoV1 to establish a persistent airway epithelial infection. This is the first report of pyroptosis in airway epithelia infected by a respiratory virus. IMPORTANCE Microbial infection of immune cells often induces pyroptosis, which is mediated by a cytosolic protein complex called the inflammasome that senses microbial pathogens and then activates the proinflammatory cytokines IL-1 and IL-18. While virus-infected airway epithelia often activate NLRP3 inflammasomes, studies to date suggest that these viruses kill the airway epithelial cells via the apoptotic or necrotic pathway; involvement of the pyroptosis pathway has not been reported previously. Here, we show for the first time that virus infection of human airway epithelia can also induce pyroptosis. Human bocavirus 1 (HBoV1), a human parvovirus, causes lower respiratory tract infections in young children. This study indicates that HBoV1 kills airway epithelial cells by activating genes that suppress apoptosis and thereby promote pyroptosis. This strategy appears to promote HBoV1 replication and may have evolved to allow HBoV1 to establish persistent infection of human airway epithelia.

scholarly journals In Vitro Modeling of Human Bocavirus 1 Infection of Polarized Primary Human Airway Epithelia

2013 ◽  
Vol 87 (7) ◽  
pp. 4097-4102 ◽  
Author(s):  
X. Deng ◽  
Z. Yan ◽  
Y. Luo ◽  
J. Xu ◽  
F. Cheng ◽  
...  
Keyword(s):  
Human Bocavirus ◽  
Airway Epithelia ◽  
In Vitro Modeling ◽  
Human Airway ◽  
Human Airway Epithelia

scholarly journals Establishment of a Reverse Genetics System for Studying Human Bocavirus in Human Airway Epithelia

2012 ◽  
Vol 8 (8) ◽  
pp. e1002899 ◽  
Author(s):  
Qinfeng Huang ◽  
Xuefeng Deng ◽  
Ziying Yan ◽  
Fang Cheng ◽  
Yong Luo ◽  
...  
Keyword(s):  
Reverse Genetics ◽  
Human Bocavirus ◽  
Airway Epithelia ◽  
Human Airway ◽  
Human Airway Epithelia

scholarly journals Characterization and Treatment of SARS-CoV-2 in Nasal and Bronchial Human Airway Epithelia

2020 ◽  
Vol 1 (4) ◽  
pp. 100059 ◽  
Author(s):  
Andrés Pizzorno ◽  
Blandine Padey ◽  
Thomas Julien ◽  
Sophie Trouillet-Assant ◽  
Aurélien Traversier ◽  
...  
Keyword(s):  
Airway Epithelia ◽  
Human Airway ◽  
Human Airway Epithelia

scholarly journals Chronic β2AR stimulation limits CFTR activation in human airway epithelia

JCI Insight ◽  
2018 ◽  
Vol 3 (4) ◽  
Author(s):  
John J. Brewington ◽  
Jessica Backstrom ◽  
Amanda Feldman ◽  
Elizabeth L. Kramer ◽  
Jessica D. Moncivaiz ◽  
...  
Keyword(s):  
Airway Epithelia ◽  
Human Airway ◽  
Human Airway Epithelia
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