Physicochemical and immunological characterization of hepatitis B virus envelope particles exclusively consisting of the entire L (pre-S1+pre-S2+S) protein

Yamada, Tesshi; Iwabuki, Hidehiko; Kanno, Takashi; Tanaka, Hiroyuki; Kawai, Tomoji; Fukuda, Hideki; Kondo, Akihiko; Seno, Masaharu; Tanizawa, Katsuyuki; Kuroda, Shinya

doi:10.1016/s0264-410x(01)00017-2

Cited by 66 publications

(35 citation statements)

References 28 publications

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“…20 After fractionation of the lyses by CsCl and sucrose gradient ultracentrifugation, L nanoparticles in the various fractions were separated by SDS-PAGE which was stained with silver reagent, as described previously. 20,22 The S-antigenic activity in the cell extracts was determined by the IMx system.…”

Section: Resultsmentioning

confidence: 99%

“…20 The whole cell extract of the recombinant yeast was fractionated with PEG 6000, and separated twice by CsCl isopycnic ultracentrifugation and once by sucrose density gradient ultracentrifugation, as described previously. 20,22 The level of the HBsAg L proteins in the yeast cell extracts were determined by silver-stained SDS-PAGE. The S-antigenic activity in the cell extracts was determined by the IMx system (Abbott Laboratories, Abbott Park, IL, USA) in conjunction with a microparticle enzyme immunoassay (MEIA) in accordance with the manufacturer's instructions.…”

Section: Methodsmentioning

confidence: 99%

“…19,21 We have succeeded in loading several different genes into the particles, and then transferring these genes into human liver cells in vitro and in vivo. In addition, L nanoparticles are easily produced by recombinant yeast cells and large-scale production of the particles has already been established, 20,22 whereas large-scale manufacturing of clinical-grade viral vectors remains a significant obstacle that hampers its clinical implementation. 23,24 It is not known whether nanoparticles could be used successfully for targeted delivery of therapeutic genes with anti-tumor activity into human liver tumors.…”

Section: Introductionmentioning

confidence: 99%

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Gene therapy of liver tumors with human liver-specific nanoparticles

et al. 2006

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The development of safe and efficient liver-specific gene delivery approaches offers new perspectives for the treatment of liver disease, in particular, liver cancer. We evaluated the therapeutic potential of hepatotropic nanoparticles for gene therapy of liver tumor. These nanoparticles do not contain a viral genome and display the hepatitis B virus L antigen, which is essential to confer hepatic specificity. It has not been shown whether a therapeutic effect could be obtained using L nanoparticles in a human liver tumor xenograft model. Rats bearing human hepatic (NuE) and non-hepatic tumors were injected with L nanoparticles containing a green fluorescent protein (GFP) expression plasmid. GFP expression was observed only in NuE-derived tumors but not in the nonhepatic tumor. The potential for treatment of liver tumors was analyzed using L nanoparticles containing the herpes simplex virus thymidine kinase gene, in conjunction with ganciclovir pro-drug administration. The growth of NuE-derived tumors in L particleinjected rats was significantly suppressed, but not of the non-hepatic tumor control. In summary, this is the first demonstration that nanoparticles could be used for delivery of therapeutic genes with anti-tumor activity into human liver tumors. This intravenous delivery system may be one of the major advantages as compared to many other viral vector systems.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Gene therapy of liver tumors with human liver-specific nanoparticles

et al. 2006

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“…When the L protein gene was expressed in Saccharomyces cerevisiae, NPs with a size of ~50 nm were overexpressed in the cells 12 and could be promptly purified by heat treatment and column chromatography. 13,14 The NP consists of about 110 molecules of L protein embedded in a unilamellar LP, displaying human liver-specific receptor outwardly.…”

mentioning

confidence: 99%

“…Furthermore, since BNCs are stable up to 70°C, 14 the phase transition temperature of LPs is estimated to be above 70°C, and BNC contains fusogenic activity in the translocation motifs of preS2 and/or S region. 23,24 We then examined whether the BNC-LP complex formation was enhanced with incubation at 70°C under acidic conditions (pH 3.0).…”

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confidence: 99%

Virosomes of hepatitis B virus envelope L proteins containing doxorubicin: synergistic enhancement of human liver-specific antitumor growth activity by radiotherapy

Liu¹,

Jung²,

Somiya³

et al. 2015

IJN

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Bionanocapsules (BNCs) are hollow nanoparticles consisting of hepatitis B virus (HBV) envelope L proteins and have been shown to deliver drugs and genes specifically to human hepatic tissues by utilizing HBV-derived infection machinery. The complex of BNCs with liposomes (LPs), the BNC–LP complexes (a LP surrounded by BNCs in a rugged spherical form), could also become active targeting nanocarriers by the BNC function. In this study, under acidic conditions and high temperature, BNCs were found to fully fuse with LPs (smooth-surfaced spherical form), deploying L proteins with a membrane topology similar to that of BNCs (ie, virosomes displaying L proteins). Doxorubicin (DOX) was efficiently encapsulated via the remote loading method at 14.2%±1.0% of total lipid weight (mean ± SD, n=3), with a capsule size of 118.2±4.7 nm and a ζ-potential of −51.1±1.0 mV (mean ± SD, n=5). When mammalian cells were exposed to the virosomes, the virosomes showed strong cytotoxicity in human hepatic cells (target cells of BNCs), but not in human colon cancer cells (nontarget cells of BNCs), whereas LPs containing DOX and DOXOVES (structurally stabilized PEGylated LPs containing DOX) did not show strong cytotoxicity in either cell type. Furthermore, the virosomes preferentially delivered DOX to the nuclei of human hepatic cells. Xenograft mice harboring either target or nontarget cell-derived tumors were injected twice intravenously with the virosomes containing DOX at a low dose (2.3 mg/kg as DOX, 5 days interval). The growth of target cell-derived tumors was retarded effectively and specifically. Next, the combination of high dose (10.0 mg/kg as DOX, once) with tumor-specific radiotherapy (3 Gy, once after 2 hours) exhibited the most effective antitumor growth activity in mice harboring target cell-derived tumors. These results demonstrated that the HBV-based virosomes containing DOX could be an effective antitumor nanomedicine specific to human hepatic tissues, especially in combination with radiotherapy.

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Bio‐nanocapsule‐based scaffold improves the sensitivity and ligand‐binding capacity of mammalian receptors on the sensor chip

Iijima

Yoshimoto

Niimi

et al. 2016

Biotechnology Journal

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Mammalian receptors are recognized as target molecules for drug discovery, and chemical libraries have been screened for both potential antagonists and agonists mainly by ligand-binding assays using immobilized receptors. A bio-nanocapsule (BNC) of approximately 30 nm that displays a tandem form of the protein A-derived immunoglobulin G (IgG) Fc-binding Z domains (denoted as ZZ-BNC) has been developed for both clustering and oriented immobilization of IgGs on the solid phase of immunosensors. In this study, human IgG1 Fc-fused vascular endothelial growth factor (VEGF) receptor was immobilized through ZZ-BNC on the sensor chip of quartz crystal microbalance (ZZ-BNC-coating). When compared with direct adsorption and protein A-coating, the sensor chip showed higher sensitivity (∽46- and ∽165-fold, respectively) and larger ligand-binding capacity (∽4- and ∽18-fold, respectively). Furthermore, the number of VEGF molecules bound to its receptor increased from 0.20 (direct adsorption) to 2.06 by ZZ-BNC-coating, strongly suggesting that ZZ-BNC reduced the steric hindrance near ligand recognition sites through oriented immobilization. Similarly, the sensitivity and ligand-binding capacity of leptin and prolactin receptors were both enhanced at a level comparable to that observed for the VEGF receptor. Thus, the combination of ZZ-BNC and Fc-fused receptors could significantly improve the function of ligand-binding assays.

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Physicochemical and immunological characterization of hepatitis B virus envelope particles exclusively consisting of the entire L (pre-S1+pre-S2+S) protein

Cited by 66 publications

References 28 publications

Gene therapy of liver tumors with human liver-specific nanoparticles

Gene therapy of liver tumors with human liver-specific nanoparticles

Virosomes of hepatitis B virus envelope L proteins containing doxorubicin: synergistic enhancement of human liver-specific antitumor growth activity by radiotherapy

Bio‐nanocapsule‐based scaffold improves the sensitivity and ligand‐binding capacity of mammalian receptors on the sensor chip

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Physicochemical and immunological characterization of hepatitis B virus envelope particles exclusively consisting of the entire L (pre-S1+pre-S2+S) protein

Cited by 66 publications

References 28 publications

Gene therapy of liver tumors with human liver-specific nanoparticles

Gene therapy of liver tumors with human liver-specific nanoparticles

Virosomes of hepatitis B virus envelope L&nbsp;proteins containing doxorubicin: synergistic enhancement of human liver-specific antitumor growth activity by radiotherapy

Bio‐nanocapsule‐based scaffold improves the sensitivity and ligand‐binding capacity of mammalian receptors on the sensor chip

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Virosomes of hepatitis B virus envelope L proteins containing doxorubicin: synergistic enhancement of human liver-specific antitumor growth activity by radiotherapy