title: Characterization of human bocavirus-like particles generated by recombinant baculoviruses.
abstract: Human bocavirus (HBoV) is a nonenveloped, single-stranded DNA virus, classified recently into the genus Bocavirus in the family Parvoviridae. A recombinant baculovirus expression system was used to express the major capsid protein VP2 of HBoV1, HBoV2, HBoV3 and HBoV4 in insect cells. A large amount of the 61-kDa VP2 capsid protein (p61) of HBoVs was generated and efficiently released into the supernatant. The capsid protein was self-assembled into 22-nm-dia. virus-like particles (VLPs) with a buoyant density of 1.30g/cm(3). The morphology of HBoVs-LPs was similar to that of the native HBoV particles, and immunogenic studies demonstrated the cross-reactivity among HBoV1, HBoV2, HBoV3 and HBoV4. When VP1 and VP2 protein of HBoV1 were co-expressed in insect cells, both proteins were detected in the same fraction after CsCl gradient centrifugation, suggesting that the VP1 protein is a minor structural protein of HBoVs. We developed an ELISA using purified VLPs as the antigen and used it to detect antibodies against HBoV1, HBoV2, HBoV3 and HBoV4. A high prevalence of antibodies against HBoVs was found in a general population of healthy Japanese, indicating that HBoVs have spread throughout Japan.
authors.affiliation: Institute of Pathogenic Microbiology, Center for Disease Control and Prevention of Guangdong Province, 160 Qunxian Road, Dashi Street, Panyu District, Guangzhou 511430, China.
authors.forename: Ling
authors.initials: L
authors.lastname: Fang
authors.affiliation: Department of Genetic Engineering, Wuhan Institute of Biological Products (WIBP) , Linjiang Avenue, Wuchang, Wuhan, 430060, China.
authors.forename: Zejun
authors.initials: Z
authors.lastname: Wang
authors.affiliation: Institute of Microbiology, Yuhang Center for Disease Control and Prevention, 930 Shijidadao Avenue, Hangzhou 311100, China.
authors.forename: Shili
authors.initials: S
authors.lastname: Song
authors.affiliation: Department of Pathology, National Institute of Infectious Diseases, Gakuen 4-7-1, Musashi-murayama 208-0011, Tokyo, Japan.
authors.forename: Michiyo
authors.initials: M
authors.lastname: Kataoka
authors.affiliation: Institute of Pathogenic Microbiology, Center for Disease Control and Prevention of Guangdong Province, 160 Qunxian Road, Dashi Street, Panyu District, Guangzhou 511430, China.
authors.forename: Changwen
authors.initials: C
authors.lastname: Ke
authors.affiliation: Department of Infectious Diseases, Hamamatsu University School of Medicine, 1-20-1 Handayama, Higashi-ku, Hamamatsu 431-3192, Japan.
authors.forename: Tetsuro
authors.initials: T
authors.lastname: Suzuki
authors.affiliation: Department of Virology II, National Institute of Infectious Diseases, Gakuen 4-7-1, Musashi-murayama 208-0011, Tokyo, Japan.
authors.forename: Takaji
authors.initials: T
authors.lastname: Wakita
authors.affiliation: Research Institute for Microbial Diseases, Osaka University, Suita 565-0781, Osaka, Japan.
authors.forename: Naokazu
authors.initials: N
authors.lastname: Takeda
authors.affiliation: Department of Virology II, National Institute of Infectious Diseases, Gakuen 4-7-1, Musashi-murayama 208-0011, Tokyo, Japan. Electronic address: litc@nih.go.jp.
authors.forename: Tian-Cheng
authors.initials: TC
authors.lastname: Li
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000914:Antibodies, Viral / Q000097:blood*; D016367:Baculoviridae / Q000235:genetics*; D036022:Capsid Proteins / Q000235:genetics / Q000378:metabolism*; D002499:Centrifugation, Density Gradient; D002648:Child; D002675:Child, Preschool; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005260:Female; D005822:Genetic Vectors; D056661:Human bocavirus / Q000235:genetics*; D006801:Humans; D007223:Infant; D007564:Japan / Q000453:epidemiology; D008297:Male; D008875:Middle Aged; D010322:Parvoviridae Infections / Q000175:diagnosis* / Q000453:epidemiology; D015995:Prevalence; D022701:Virosomes / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D055815:Young Adult
chemical_list: D000914:Antibodies, Viral; D036022:Capsid Proteins; D022701:Virosomes
keywords: HBoV1–4; Human bocaviruses; Insect cells Tn5 and Sf9; Recombinant baculovirus; VLP; Virus-like particles