Human papillomavirus (HPV) type 11 recombinant virus-like particles induce the formation of neutralizing antibodies and detect HPV-specific antibodies in human sera

Rose, Robert C.; Reichman, Richard C.; Bonnez, William

doi:10.1099/0022-1317-75-8-2075

Cited by 147 publications

(99 citation statements)

References 21 publications

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“…Antibodies-Rabbit polyclonal antiserum was raised against HPV16 L1 capsids as described (27,28). Murine antibodies raised against Kap ␣2/Rch-1 and Kap ␤2/transportin were from Transduction Laboratories; a mouse monoclonal antibody to Kap ␤1/p97 was from Affinity Bioreagents, Inc., and the goat anti-GST antibody was from Amersham Biosciences.…”

Section: Methodsmentioning

confidence: 99%

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Nuclear Import Strategies of High Risk HPV16 L1 Major Capsid Protein

Nelson¹,

Rose²,

Moroianu³

2002

Journal of Biological Chemistry

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During the late phase of human papillomavirus (HPV) infection, the L1 major capsid proteins enter the nuclei of host epithelial cells and, together with the L2 minor capsid proteins, assemble the replicated viral DNA into virions. We investigated the nuclear import of the L1 major capsid protein of high risk HPV16. When digitonin-permeabilized HeLa cells were incubated with HPV16 L1 capsomeres, the L1 protein was imported into the nucleus in a receptor-mediated manner. HPV16 L1 capsomeres formed complexes with Kap ␣2␤1 heterodimers via interaction with Kap ␣2. Accordingly, nuclear import of HPV16 L1 capsomeres was mediated by Kap ␣2␤1 heterodimers, required RanGDP and free GTP, and was independent of GTP hydrolysis. Remarkably, HPV16 L1 capsomeres also interacted with Kap ␤2 and binding of RanGTP to Kap ␤2 did not dissociate the HPV16 L1⅐Kap ␤2 complex. Significantly, HPV16 L1 capsomeres inhibited the nuclear import of Kap ␤2 and of a Kap ␤2-specific M9-containing cargo. These data suggest that, during the productive stage of infection, while the HPV16 L1 major capsid protein enters the nucleus via the Kap ␣2␤1-mediated pathway to assemble the virions, it also inhibits the Kap ␤2-mediated nuclear import of host hnRNP A1 protein and, in this way, favors virion formation.Human papillomaviruses (HPVs) 1 are thought to be the primary causative agent in more than 90% of all cervical cancers, with HPV16 being the type most frequently found in these tumors. Approximately 500,000 new cases of cervical cancer are identified each year globally with nearly 300,000 deaths annually. About 85 genotypically distinct HPV genotypes have been isolated and characterized, with roughly half infecting the skin and the other half preferentially infecting oral/anogenital mucosal epithelial tissues. Mucosal HPVs have demonstrated varying degrees of oncogenic potential; high risk HPVs, such as types 16, 18, 31, and 45, are frequently detected in invasive cervical carcinomas, whereas the low risk types, such as types 6 and 11, are more often associated with benign exophytic condylomas (1).HPVs are small, nonenveloped, icosahedral DNA viruses that replicate in the nucleus of squamous epithelial cells. The virion particles (52-55 nm in diameter) consist of a single molecule comprising 8 kb of double-stranded circular DNA contained within a spherical capsid composed of 72 homopentameric L1 capsomeres and of L2 minor capsid protein. The number of L2 molecules per capsid has been estimated at 36 (2) or 12 (3). L1 protein is capable of self-assembly both in vivo and in vitro into capsid-like structures, referred to as virus-like particles (4 -9). L1 is stable in two oligomeric configurations: homopentameric capsomeres and capsids composed of 72 capsomeres. The capsids can be disassembled into capsomeres quantitatively by an agent that reduces disulfide bonds and reassembled by removing the reducing agent (10, 11). The L1 capsid proteins of HPVs seem to enter the nucleus of host cells twice during the virus life cycle: immediately after the vi...

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Section: Methodsmentioning

confidence: 99%

“…Preparation of Recombinant HPV16 L1 Capsids and CapsomeresThe L1 capsids of HPV16 and HPV45 were generated in insect cells, purified as described (27,28), and stored at 4°C until use. Purity and lack of proteolytic degradation of the L1 proteins were always checked by SDS-PAGE, Coomassie Blue staining, and immunoblotting prior to use.…”

Section: Methodsmentioning

confidence: 99%

Nuclear Import Strategies of High Risk HPV16 L1 Major Capsid Protein

Nelson¹,

Rose²,

Moroianu³

2002

Journal of Biological Chemistry

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“…These vaccines are based on viruslike particles (VLPs), which assemble spontaneously when the papillomavirus major capsid protein, L1, is recombinantly expressed either alone or together with L2, the minor capsid protein (Hofmann et al, 1995;Kirnbauer et al, 1992Kirnbauer et al, , 1993Neeper et al, 1996;Rose et al, 1993;Volpers et al, 1994;Zhou et al, 1991). VLPs are morphologically and immunologically similar to native virions (Christensen et al, 1994;Kirnbauer et al, 1992;Neeper et al, 1996;Rose et al, 1993), can induce neutralizing antibody responses Christensen et al, 1994Christensen et al, , 1996bLowe et al, 1997;Ludmerer et al, 2000;Pastrana et al, 2001;Roden et al, 1997;Rose et al, 1994;Yeager et al, 2000) and in animal models, can protect against viral disease (Breitburd et al, 1995;Christensen et al, 1996c;Jansen et al, 1995;Kirnbauer et al, 1996;Suzich et al, 1995).…”

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confidence: 99%

Human papillomavirus type 6 virus-like particles present overlapping yet distinct conformational epitopes

Wang

Cook

Lee

et al. 2003

Journal of General Virology

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The epitope for a human papillomavirus (HPV) type 6 conformation-dependent, neutralizing monoclonal antibody (mAb) was partially mapped using HPV L1 recombinant virus-like particles (VLPs). The mAb H6.J54 is cross-reactive with the closely related HPV types 6 and 11. By making HPV-6-like amino acid substitutions in the cottontail rabbit papillomavirus (CRPV) major capsid protein L1, we were able to transfer H6.J54 binding activity into a CRPV/HPV-6 hybrid L1 protein.Full binding activity was achieved with only nine amino acid changes and identified a region centred on the HPV-6 residues 49-54. This region has previously been shown to be a critical part of HPV-6 type-specific epitopes. Fine mapping of the region by scanning a series of alanine substitution mutations showed that in HPV-6 VLPs this type-common epitope overlaps HPV-6 type-specific epitopes.

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“…In a study by Rose et al, rabbits were immunised with VLPs of HPV 11 produced in Sf9 cells (expressed by a recombinant baculovirus) and purified by caesium chloride density gradient centrifugation (Ref. 79). Serum was then tested for neutralising antibodies in the athymic mouse model of HPV 11 infection.…”

Section: Eliciting Humoral Responsesmentioning

confidence: 99%