Establishment of a reverse genetics system for studying human bocavirus in human airway epithelia

Qinfeng Huang; Xuefeng Deng; Ziying Yan; Fang Cheng; Yong Luo; Weiran Shen; Diana C M Lei-Butters; Aaron Yun Chen; Yi Li; Liang Tang; Maria Söderlund-Venermo; John F Engelhardt; Jianming Qiu

doi:10.1371/journal.ppat.1002899

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Establishment of a reverse genetics system for studying human bocavirus in human airway epithelia

Journal article

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Establishment of a reverse genetics system for studying human bocavirus in human airway epithelia

Qinfeng Huang, Xuefeng Deng, Ziying Yan, Fang Cheng, Yong Luo, Weiran Shen, Diana C M Lei-Butters, Aaron Yun Chen, Yi Li, Liang Tang, …

PLoS pathogens, Vol.8(8), pp.e1002899-e1002899

2012

DOI: 10.1371/journal.ppat.1002899

PMCID: PMC3431310

PMID: 22956907

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Abstract

Human bocavirus 1 (HBoV1) has been identified as one of the etiological agents of wheezing in young children with acute respiratory-tract infections. In this study, we have obtained the sequence of a full-length HBoV1 genome (including both termini) using viral DNA extracted from a nasopharyngeal aspirate of an infected patient, cloned the full-length HBoV1 genome, and demonstrated DNA replication, encapsidation of the ssDNA genome, and release of the HBoV1 virions from human embryonic kidney 293 cells. The HBoV1 virions generated from this cell line-based production system exhibits a typical icosahedral structure of approximately 26 nm in diameter, and is capable of productively infecting polarized primary human airway epithelia (HAE) from the apical surface. Infected HAE showed hallmarks of lung airway-tract injury, including disruption of the tight junction barrier, loss of cilia and epithelial cell hypertrophy. Notably, polarized HAE cultured from an immortalized airway epithelial cell line, CuFi-8 (originally derived from a cystic fibrosis patient), also supported productive infection of HBoV1. Thus, we have established a reverse genetics system and generated the first cell line-based culture system for the study of HBoV1 infection, which will significantly advance the study of HBoV1 replication and pathogenesis.

Transfection

Cell Line

Reverse Genetics

Parvoviridae Infections - virology

Genome, Viral - genetics

Humans

Molecular Sequence Data

Human bocavirus - isolation & purification

Sequence Analysis, DNA

Epithelium - virology

DNA, Viral - chemistry

Respiratory System - virology

Human bocavirus - genetics

Human bocavirus - physiology

Virus Replication

Base Sequence

Epithelial Cells - virology

Female

Respiratory Tract Infections - virology

DNA, Viral - genetics

Inverted Repeat Sequences

Details

Title: Subtitle: Establishment of a reverse genetics system for studying human bocavirus in human airway epithelia
Creators: Qinfeng Huang - Department of Microbiology, Molecular Genetics and Immunology, University of Kansas Medical Center, Kansas City, Kansas, USA
Xuefeng Deng
Ziying Yan
Fang Cheng
Yong Luo
Weiran Shen
Diana C M Lei-Butters
Aaron Yun Chen
Yi Li
Liang Tang
Maria Söderlund-Venermo
John F Engelhardt
Jianming Qiu
Resource Type: Journal article
Publication Details: PLoS pathogens, Vol.8(8), pp.e1002899-e1002899
DOI: 10.1371/journal.ppat.1002899
PMID: 22956907
PMCID: PMC3431310
NLM abbreviation: PLoS Pathog
ISSN: 1553-7366
eISSN: 1553-7374
Publisher: Public Library of Science; United States
Grant note: R01 HL108902 / NHLBI NIH HHS R21 AI085236 / NIAID NIH HHS R01 AI070723 / NIAID NIH HHS R56 AI070723 / NIAID NIH HHS P30 DK054759 / NIDDK NIH HHS
Language: English
Date published: 2012
Academic Unit: Roy J. Carver Department of Biomedical Engineering; Anatomy and Cell Biology; Radiation Oncology; Internal Medicine
Record Identifier: 9984025469402771

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