Merck
  • Home
  • Search Results
  • Hepatitis E virus egress depends on the exosomal pathway, with secretory exosomes derived from multivesicular bodies.

Hepatitis E virus egress depends on the exosomal pathway, with secretory exosomes derived from multivesicular bodies.

The Journal of general virology (2014-06-28)
Shigeo Nagashima, Suljid Jirintai, Masaharu Takahashi, Tominari Kobayashi, Tanggis, Tsutomu Nishizawa, Tom Kouki, Takashi Yashiro, Hiroaki Okamoto
ABSTRACT

Our previous studies indicated that hepatitis E virus (HEV) forms membrane-associated particles in the cytoplasm, most likely by budding into intracellular vesicles, and requires the multivesicular body (MVB) pathway to release virus particles, and the released HEV particles with a lipid membrane retain the trans-Golgi network protein 2 on their surface. To examine whether HEV utilizes the exosomal pathway to release the virus particles, we analysed whether the virion release from PLC/PRF/5 cells infected with genotype 3 HEV (strain JE03-1760F) is affected by treatment with bafilomycin A1 or GW4869, or by the introduction of a small interfering RNA (siRNA) against Rab27A or Hrs. The extracellular HEV RNA titre was increased by treatment with bafilomycin A1, but was decreased by treatment with GW4869. The relative levels of virus particles released from cells depleted of Rab27A or Hrs were decreased to 16.1 and 11.5 %, respectively, of that released from cells transfected with negative control siRNA. Electron microscopic observations revealed the presence of membrane-associated virus-like particles with a diameter of approximately 50 nm within the MVB, which possessed internal vesicles in infected cells. Immunoelectron microscopy showed positive immunogold staining for the HEV ORF2 protein on the intraluminal vesicles within the MVB. Additionally, immunofluorescence analysis indicated the triple co-localization of the ORF2, ORF3 and CD63 proteins in the cytoplasm, as specific loculated signals, supporting the presence of membrane-associated HEV particles within the MVB. These findings indicate that membrane-associated HEV particles are released together with internal vesicles through MVBs by the cellular exosomal pathway.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
二甲基亚砜, Hybri-Max, sterile-filtered, BioReagent, suitable for hybridoma, ≥99.7%
Sigma-Aldrich
二甲基亚砜, anhydrous, ≥99.9%
Sigma-Aldrich
二甲基亚砜, for molecular biology
Sigma-Aldrich
二甲基亚砜, ACS reagent, ≥99.9%
Sigma-Aldrich
二甲基亚砜, sterile-filtered, BioPerformance Certified, meets EP, USP testing specifications, suitable for hybridoma
Sigma-Aldrich
氯化钠, BioXtra, ≥99.5% (AT)
Sigma-Aldrich
二甲基亚砜, suitable for HPLC, ≥99.7%
Sigma-Aldrich
二甲基亚砜, ReagentPlus®, ≥99.5%
Sigma-Aldrich
二甲基亚砜, ≥99.5% (GC), suitable for plant cell culture
Sigma-Aldrich
氯化钠, for molecular biology, DNase, RNase, and protease, none detected, ≥99% (titration)
Sigma-Aldrich
DAPI, for nucleic acid staining
Sigma-Aldrich
氯化钠 溶液, 5 M in H2O, BioReagent, for molecular biology, suitable for cell culture
Sigma-Aldrich
氯化钠, BioReagent, suitable for cell culture, suitable for insect cell culture, suitable for plant cell culture, ≥99%
Sigma-Aldrich
氯化钠 溶液, 0.9% in water, BioXtra, suitable for cell culture
SAFC
氯化钠 溶液, 5 M
Sigma-Aldrich
氯化钠 溶液, 5 M
Sigma-Aldrich
二甲基亚砜, meets EP testing specifications, meets USP testing specifications
Sigma-Aldrich
二甲基亚砜, puriss. p.a., ACS reagent, ≥99.9% (GC)
Sigma-Aldrich
氯化钠, meets analytical specification of Ph. Eur., BP, USP, 99.0-100.5%
Sigma-Aldrich
二甲基亚砜, BioUltra, for molecular biology, ≥99.5% (GC)
Sigma-Aldrich
氯化钠, BioUltra, for molecular biology, ≥99.5% (AT)
Supelco
氯化钠, Pharmaceutical Secondary Standard; Certified Reference Material
Sigma-Aldrich
二甲基亚砜, PCR Reagent
Sigma-Aldrich
氯化钠 溶液, 0.85%
Sigma-Aldrich
氯化钠 溶液, BioUltra, for molecular biology, ~5 M in H2O
Supelco
氯化钠, certified reference material for titrimetry, certified by BAM, ≥99.5%
Sigma-Aldrich
氯化钠, 99.999% trace metals basis
Sigma-Aldrich
氯化钠, BioPerformance Certified, ≥99% (titration), suitable for insect cell culture, suitable for plant cell culture
Supelco
二甲基亚砜, analytical standard
Sigma-Aldrich
氯化钠, tested according to Ph. Eur.