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Expression of E1E2 on hepatitis C RNA-containing particles released from primary cultured human hepatocytes derived from infected cirrhotic livers
- Source :
- Intervirology, Intervirology, Karger, 2011, 54 (1), pp.1-9. ⟨10.1159/000318886⟩
- Publication Year :
- 2011
- Publisher :
- HAL CCSD, 2011.
-
Abstract
- Objective: To determine whether liver-derived hepatitis C RNA-containing particles express the E1E2 discontinuous antigenic determinant defined by unique monoclonal antibody (mAb) D32.10 which recognizes three highly conserved segments in E1 (aa297–306) and E2 (aa480–494 and aa613–621) envelope glycoproteins. Methods: Human hepatocytes were isolated from HCV-infected cirrhotic explanted livers. The liver-derived hepatitis C virus (HCV) particles released from three distinct cultures (genotypes 1b and 2b) were characterized. HCV RNA+ was quantified by real-time RT-PCR. The E1E2 antigenic activity was assessed by indirect ELISA and immunoblotting using D32.10. The density distributions of HCV RNA and E1E2 antigen were determined by isopycnic sucrose density gradients. HCV E1E2, E2 and core antigens were detected in the cells by immunochemical staining. Results: Liver-derived HCV particles contained HCV RNA (106–107 copies/mg of protein) and core proteins and expressed the E1E2/D32.10 epitope. HCV RNA and E1E2 cosedimented between 1.15 and 1.25 g/ml in sucrose gradients. Moreover, the mAb D32.10 detected E1E2 by immunostaining in HCV-infected hepatocytes in parallel with E2 and core antigens. Conclusion: Our results provide evidence that the mAb D32.10 recognizes E1E2 envelope complexes expressed in the cell cytoplasm and on the surface of HCV RNA-containing particles released from short-term cultures of in vivo infected hepatocytes.
- Subjects :
- Liver Cirrhosis
Hepacivirus
Centrifugation, Isopycnic
medicine.disease_cause
Epitope
MESH: Antibodies, Monoclonal
MESH: Hepatocytes
Epitopes
MESH: Gene Expression Regulation, Viral
0302 clinical medicine
Viral Envelope Proteins
MESH: Hepacivirus
Cells, Cultured
chemistry.chemical_classification
0303 health sciences
MESH: Peptides
Antibodies, Monoclonal
Hepatitis C
3. Good health
MESH: Centrifugation, Isopycnic
Infectious Diseases
MESH: RNA, Viral
RNA, Viral
030211 gastroenterology & hepatology
MESH: Liver Cirrhosis
MESH: Cells, Cultured
Gene Expression Regulation, Viral
MESH: Epitopes
medicine.drug_class
Hepatitis C virus
[SDV.CAN]Life Sciences [q-bio]/Cancer
Biology
Monoclonal antibody
03 medical and health sciences
Antigen
In vivo
Virology
medicine
Humans
030304 developmental biology
MESH: Hepatitis C
MESH: Humans
Molecular biology
chemistry
Cytoplasm
MESH: Viral Envelope Proteins
Hepatocytes
Glycoprotein
Peptides
Immunostaining
Subjects
Details
- Language :
- English
- ISSN :
- 03005526 and 14230100
- Database :
- OpenAIRE
- Journal :
- Intervirology, Intervirology, Karger, 2011, 54 (1), pp.1-9. ⟨10.1159/000318886⟩
- Accession number :
- edsair.doi.dedup.....47d7a83e2c9ae6d332571ebc5e12aff6