Hepatitis C virus particles of different density in the blood of chronically infected immunocompetent and immunodeficient patients: Implications for virus clearance by antibody

Pumeechockchai, Wanna; Bevitt, Debra J.; Agarwal, K. N.; Petropoulou, Theoni; Langer, Barbara; Belohradsky, Bernd H.; Bassendine, MF; Toms, GL

doi:10.1002/jmv.10208

Cited by 46 publications

(55 citation statements)

References 23 publications

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“…Our immunoprecipitation analysis showed that the LVPs contain both ApoB and ApoE. ApoB is found on all low-density lipoproteins, but the presence of ApoE is characteristic of VLDL We have found that HCV core, as well as glycoproteins E1 and E2, is present in fractions with densities below 1.08 g/ml from liver S6b (42) (Fig. 6), and this has allowed us to seek antibodies capable of immunoprecipitating these viral proteins.…”

Section: Discussionmentioning

confidence: 85%

“…A sample of 100 l from patient S6 or 1 ml from patients S7 to S9 was mixed with NaBr solution with a density of 1.063 g/ml and centrifuged at 146,000 ϫ g av (g av is the average centrifugal force) in a Beckman Type 50 Ti fixed-angle rotor (42). After sequential ultracentrifugation, three samples of 1 ml each were harvested from low-density ( Ͻ 1.063 g/ml), intermediate-density (1.063 g/ml Ͻ Ͻ 1.21 g/ml), and high-density ( Ͻ 1.21 g/ml) fractions.…”

Section: Methodsmentioning

confidence: 99%

“…RNA was extracted from 100 l of each gradient fraction using a QIAamp viral-RNA kit (QIAGEN, Crawley, United Kingdom). Reverse transcription (RT) and PCR were performed with two oligonucleotide primers corresponding to bases 29 to 46 (sense) and 251 to 268 (antisense) of the 5Ј noncoding region of HCV as described previously (42). The 240-bp PCR product was analyzed on 2% agarose gels, and the intensities of bands were semiquantitated with an AlphaImager 2200 digitalimage camera (Flowgen, Ashby de la Zouch, United Kingdom).…”

Section: Methodsmentioning

confidence: 99%

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Association between Hepatitis C Virus and Very-Low-Density Lipoprotein (VLDL)/LDL Analyzed in Iodixanol Density Gradients

Nielsen¹,

Bassendine²,

Burt

et al. 2006

J Virol

309

324

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Hepatitis C virus (HCV) RNA circulates in the blood of persistently infected patients in lipoviroparticles (LVPs), which are heterogeneous in density and associated with host lipoproteins and antibodies. The variability and lability of these virus-host complexes on fractionation has hindered our understanding of the structure of LVP and determination of the physicochemical properties of the HCV virion. In this study, HCV from an antibody-negative immunodeficient patient was analyzed using three fractionation techniques, NaBr gradients, isotonic iodixanol, and sucrose gradient centrifugation. Iodixanol gradients were shown to best preserve host lipoprotein-virus complexes, and all HCV RNA was found at densities below 1.13 g/ml, with the majority at low density, <1.08 g/ml. Immunoprecipitation with polyclonal antibodies against human ApoB and ApoE precipitated 91.8% and 95.0% of HCV with low density, respectively, suggesting that host lipoprotein is closely associated with HCV in a particle resembling VLDL. Immunoprecipitation with antibodies against glycoprotein E2 precipitated 25% of HCV with low density, providing evidence for the presence of E2 in LVPs. Treatment of serum with 0.5% deoxycholic acid in the absence of salt produced HCV with a density of 1.12 g/ml and a sedimentation coefficient of 215S. The diameters of these particles were calculated as 54 nm. Treatment of serum with 0.18% NP-40 produced HCV with a density of 1.18 g/ml, a sedimentation coefficient of 180S, and a diameter of 42 nm. Immunoprecipitation analysis showed that ApoB remained associated with HCV after treatment of serum with deoxycholic acid or NP-40, whereas ApoE was removed from HCV with these detergents.

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Section: Discussionmentioning

confidence: 85%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Association between Hepatitis C Virus and Very-Low-Density Lipoprotein (VLDL)/LDL Analyzed in Iodixanol Density Gradients

Nielsen¹,

Bassendine²,

Burt

et al. 2006

J Virol

309

324

View full text Add to dashboard Cite

show abstract

“…It has been suggested that, in chronically infected patients, HCV circulates as low-density lipoprotein virus particles (1,40), with various density profiles depending on the stage of the infection at which the sample was obtained (9,43). The differences in density and infectivity have been attributed to the presence of host lipoproteins and antibodies bound to the circulating viral particles (22,43).…”

mentioning

confidence: 99%

Differential Biophysical Properties of Infectious Intracellular and Secreted Hepatitis C Virus Particles

Gastaminza

Kapadia

Chisari

2006

J Virol

215

232

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The recent development of a cell culture infection model for hepatitis C virus (HCV) permits the production of infectious particles in vitro. In this report, we demonstrate that infectious particles are present both within the infected cells and in the supernatant. Kinetic analysis indicates that intracellular particles constitute precursors of the secreted infectious virus. Ultracentrifugation analyses indicate that intracellular infectious viral particles are similar in size (ϳ65 to 70 nm) but different in buoyant density (ϳ1.15 to 1.20 g/ml) from extracellular particles (ϳ1.03 to 1.16 g/ml). These results indicate that infectious HCV particles are assembled intracellularly and that their biochemical composition is altered during viral egress.Hepatitis C virus (HCV) is a major cause of chronic hepatitis worldwide. Approximately 3% of the human population is infected, and more than 80% of all HCV infections progress to chronicity, ultimately leading to fibrosis, cirrhosis, and hepatocellular carcinoma (24). There is no vaccine against HCV, and the most widely used therapy involves the administration of type I interferon (␣2A) combined with ribavirin. However, this treatment strategy is toxic and has been shown to be ineffective in a significant proportion of the cases (41).HCV is a member of the Flaviviridae family and the sole member of the genus Hepacivirus (34). HCV is an enveloped virus with a single-strand positive RNA genome that codes for a unique polyprotein of approximatively 3,000 amino acids (11,12). A single open reading frame is flanked by 5Ј and 3Ј untranslated regions that contain RNA sequences essential for RNA translation and replication, respectively (17,18,23). The translation of the single open reading frame is driven by an internal ribosomal entry site sequence present within the 5Ј untranslated region (23), and the resulting polyprotein is processed by cellular and viral proteases into its individual components (reviewed in reference 42). The E1, E2, and core structural proteins are assembled into particles (3, 4), but are not essential for viral RNA replication or translation. The NS2, NS3, NS4A, NS4B, NS5A, and NS5B nonstructural proteins constitute the viral components necessary for efficient viral RNA replication, although NS2 is dispensable for this function (5, 33). In the linear sequence of the polyprotein, the structural proteins are separated from the nonstructural proteins by a small hydrophobic protein, p7 (29), whose function remains unknown but that has the potential to form ion channels (19). Viral proteins are localized in the cytoplasm, and it is assumed, by analogy with other members of the Flaviviridae family, that the entire life cycle of the virus is exclusively cytoplasmic (32).

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“…10 The subjects' clinical presentation was compared to the differential HCV density fractions. Baseline and sequential liver function tests were performed and five out of six patients proceeded to liver biopsy for clinical purposes.…”

mentioning

confidence: 99%

Hepatitis C Virus Heterogeneity: Lipoprotein and Immunoglobulin Binding and Clinical Status

Adler

Biddle

Beswick

et al. 2013

Journal of Clinical and Experimental Hepatology

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Hepatitis C virus particles of different density in the blood of chronically infected immunocompetent and immunodeficient patients: Implications for virus clearance by antibody

Cited by 46 publications

References 23 publications

Association between Hepatitis C Virus and Very-Low-Density Lipoprotein (VLDL)/LDL Analyzed in Iodixanol Density Gradients

Association between Hepatitis C Virus and Very-Low-Density Lipoprotein (VLDL)/LDL Analyzed in Iodixanol Density Gradients

Differential Biophysical Properties of Infectious Intracellular and Secreted Hepatitis C Virus Particles

Hepatitis C Virus Heterogeneity: Lipoprotein and Immunoglobulin Binding and Clinical Status

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