title: Establishment of a reverse genetics system for studying human bocavirus in human airway epithelia.
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.
authors.affiliation: Department of Microbiology, Molecular Genetics and Immunology, University of Kansas Medical Center, Kansas City, Kansas, USA.
authors.forename: Qinfeng
authors.initials: Q
authors.lastname: Huang
authors.forename: Xuefeng
authors.initials: X
authors.lastname: Deng
authors.forename: Ziying
authors.initials: Z
authors.lastname: Yan
authors.forename: Fang
authors.initials: F
authors.lastname: Cheng
authors.forename: Yong
authors.initials: Y
authors.lastname: Luo
authors.forename: Weiran
authors.initials: W
authors.lastname: Shen
authors.forename: Diana C M
authors.initials: DC
authors.lastname: Lei-Butters
authors.forename: Aaron Yun
authors.initials: AY
authors.lastname: Chen
authors.forename: Yi
authors.initials: Y
authors.lastname: Li
authors.forename: Liang
authors.initials: L
authors.lastname: Tang
authors.forename: Maria
authors.initials: M
authors.lastname: Söderlund-Venermo
authors.forename: John F
authors.initials: JF
authors.lastname: Engelhardt
authors.forename: Jianming
authors.initials: J
authors.lastname: Qiu
mesh_terms: D001483:Base Sequence; D002460:Cell Line; D004279:DNA, Viral / Q000737:chemistry / Q000235:genetics; D004847:Epithelial Cells / Q000821:virology*; D004848:Epithelium / Q000821:virology; D005260:Female; D016679:Genome, Viral / Q000235:genetics; D056661:Human bocavirus / Q000235:genetics / Q000302:isolation & purification / Q000502:physiology*; D006801:Humans; D055029:Inverted Repeat Sequences; D008969:Molecular Sequence Data; D010322:Parvoviridae Infections / Q000821:virology*; D012137:Respiratory System / Q000821:virology; D012141:Respiratory Tract Infections / Q000821:virology*; D059386:Reverse Genetics; D017422:Sequence Analysis, DNA; D014162:Transfection; D014779:Virus Replication
chemical_list: D004279:DNA, Viral