Journal article
Human Bocavirus 1 Infection of Well-Differentiated Human Airway Epithelium
Current protocols in microbiology (Online), Vol.58(1), pp.e107-e107
09/2020
DOI: 10.1002/cpmc.107
PMCID: PMC7422954
PMID: 32639683
Abstract
Human bocavirus 1 (HBoV1) is a small DNA virus that belongs to the Bocaparvovirus genus of the Parvoviridae family. HBoV1 is a common respiratory pathogen that causes mild to life-threatening acute respiratory tract infections in children and immunocompromised individuals, infecting both the upper and lower respiratory tracts. HBoV1 infection causes death of airway epithelial cells, resulting in airway injury and inflammation. In vitro, HBoV1 only infects well-differentiated (polarized) human airway epithelium cultured at an air-liquid interface (HAE-ALI), but not any dividing human cells. A full-length HBoV1 genome of 5543 nucleotides has been cloned from DNA extracted from a human nasopharyngeal swab into a plasmid called HBoV1 infectious clone pIHBoV1. Transfection of pIHBoV1 replicates efficiently in human embryonic kidney 293 (HEK293) cells and produces virions that are highly infectious. This article describes protocols for production of HBoV1 in HEK293 cells, generation of HAE-ALI cultures, and infection with HBoV1 in HAE-ALI. © 2020 Wiley Periodicals LLC. Basic Protocol 1: Human bocavirus 1 production in HEK293 cells Support Protocol 1: HEK293 cell culture and transfection Support Protocol 2: Quantification of human bocavirus 1 using real-time quantitative PCR Basic Protocol 2: Differentiation of human airway cells at an air-liquid interface Support Protocol 3: Expansion of human airway epithelial cell line CuFi-8 Support Protocol 4: Expansion of human airway basal cells Support Protocol 5: Coating of plastic dishes and permeable membranes of inserts Support Protocol 6: Transepithelial electrical resistance measurement Basic Protocol 3: Human bocavirus 1 infection in human airway epithelium cultured at an air-liquid interface Support Protocol 7: Isolation of infected human airway epithelium cells from inserts Basic Protocol 4: Transduction of airway basal cells with lentiviral vector.
Details
- Title: Subtitle
- Human Bocavirus 1 Infection of Well-Differentiated Human Airway Epithelium
- Creators
- Ziying Yan - University of IowaXuefeng Deng - University of Kansas Medical CenterJianming Qiu - University of Kansas Medical Center
- Resource Type
- Journal article
- Publication Details
- Current protocols in microbiology (Online), Vol.58(1), pp.e107-e107
- DOI
- 10.1002/cpmc.107
- PMID
- 32639683
- PMCID
- PMC7422954
- NLM abbreviation
- Curr Protoc Microbiol
- ISSN
- 1934-8525
- eISSN
- 1934-8533
- Grant note
- R01 AI150877 / NIAID NIH HHS R21 AI139572 / NIAID NIH HHS P30 DK054759 / NIDDK NIH HHS
- Language
- English
- Date published
- 09/2020
- Academic Unit
- Anatomy and Cell Biology
- Record Identifier
- 9984284326702771
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