Molecular characterization of intrahepatic and extrahepatic hepatitis B virus (HBV) reservoirs in patients on suppressive antiviral therapy

[Guest guest]

http://onlinelibrary.wiley.com/doi/10.1111/j.1365-2893.2010.01321.x/abstract

Molecular characterization of intrahepatic and extrahepatic hepatitis B virus

(HBV) reservoirs in patients on suppressive antiviral therapy

C. S. Coffin1, P. M. Mulrooney-Cousins2, M. G. s3, G. van Marle4, J. P.

5, T. I. Michalak2, N. A. Terrault3

Article first published online: 8 JUL 2010

DOI: 10.1111/j.1365-2893.2010.01321.x

© 2010 Blackwell Publishing Ltd

Issue

Journal of Viral Hepatitis

Volume 18, Issue 6, pages 415–423, June 2011

Summary.  The hepatitis B virus (HBV) replicates via an error-prone reverse

transcriptase generating potential drug-resistant quasispecies. The degree of

HBV variability in liver vs peripheral blood mononuclear cells (PBMC) in

patients on long-term suppressive antivirals is unclear. We characterized HBV

replication, drug resistance and molecular diversity in patients with plasma HBV

DNA undetectable by clinical assays. Explant liver (n = 9), PBMC (n = 6) and

plasma (n = 7) from nine such patients undergoing liver transplantation were

evaluated for HBV genomes by sensitive PCR/nucleic acid hybridization assay.

Cases with HBV DNA in liver and PBMC were tested for covalently closed circular

DNA (HBV cccDNA). HBV polymerase (P) amplicons were cloned, sequenced and both P

and overlapping surface (S) gene sequences were analysed. HBV DNA was detected

in 43% (3/7) of plasma, 100% (9/9) of liver and 83% (5/6) of PBMC samples. HBV

cccDNA was detected in all liver and one PBMC sample. Four patients had a

clinical diagnosis of resistance. HBV P gene sequencing revealed 100% wild type

(wt) in plasma (2/2), 83% wt in PBMC (5/6) but livers of 3/9 (33%) contained wt

and 6/9 (66%) carried resistance to lamivudine and/or adefovir. The translated S

gene revealed no changes affecting HBV antigenicity. Sequences from livers with

antiviral resistant mutants revealed greater interpatient quasispecies

diversity. Despite apparent HBV suppression, the liver continues to support HBV

replication and extrahepatic HBV can be detected. PBMC may be a sanctuary for wt

virus during antiviral therapy, while the liver harbours more drug-resistant

viruses. Drug resistance correlates with intrahepatic viral diversity.

[Guest guest]

http://onlinelibrary.wiley.com/doi/10.1111/j.1365-2893.2010.01321.x/abstract

Molecular characterization of intrahepatic and extrahepatic hepatitis B virus

(HBV) reservoirs in patients on suppressive antiviral therapy

C. S. Coffin1, P. M. Mulrooney-Cousins2, M. G. s3, G. van Marle4, J. P.

5, T. I. Michalak2, N. A. Terrault3

Article first published online: 8 JUL 2010

DOI: 10.1111/j.1365-2893.2010.01321.x

© 2010 Blackwell Publishing Ltd

Issue

Journal of Viral Hepatitis

Volume 18, Issue 6, pages 415–423, June 2011

Summary.  The hepatitis B virus (HBV) replicates via an error-prone reverse

transcriptase generating potential drug-resistant quasispecies. The degree of

HBV variability in liver vs peripheral blood mononuclear cells (PBMC) in

patients on long-term suppressive antivirals is unclear. We characterized HBV

replication, drug resistance and molecular diversity in patients with plasma HBV

DNA undetectable by clinical assays. Explant liver (n = 9), PBMC (n = 6) and

plasma (n = 7) from nine such patients undergoing liver transplantation were

evaluated for HBV genomes by sensitive PCR/nucleic acid hybridization assay.

Cases with HBV DNA in liver and PBMC were tested for covalently closed circular

DNA (HBV cccDNA). HBV polymerase (P) amplicons were cloned, sequenced and both P

and overlapping surface (S) gene sequences were analysed. HBV DNA was detected

in 43% (3/7) of plasma, 100% (9/9) of liver and 83% (5/6) of PBMC samples. HBV

cccDNA was detected in all liver and one PBMC sample. Four patients had a

clinical diagnosis of resistance. HBV P gene sequencing revealed 100% wild type

(wt) in plasma (2/2), 83% wt in PBMC (5/6) but livers of 3/9 (33%) contained wt

and 6/9 (66%) carried resistance to lamivudine and/or adefovir. The translated S

gene revealed no changes affecting HBV antigenicity. Sequences from livers with

antiviral resistant mutants revealed greater interpatient quasispecies

diversity. Despite apparent HBV suppression, the liver continues to support HBV

replication and extrahepatic HBV can be detected. PBMC may be a sanctuary for wt

virus during antiviral therapy, while the liver harbours more drug-resistant

viruses. Drug resistance correlates with intrahepatic viral diversity.

[Guest guest]

http://onlinelibrary.wiley.com/doi/10.1111/j.1365-2893.2010.01321.x/abstract

Molecular characterization of intrahepatic and extrahepatic hepatitis B virus

(HBV) reservoirs in patients on suppressive antiviral therapy

C. S. Coffin1, P. M. Mulrooney-Cousins2, M. G. s3, G. van Marle4, J. P.

5, T. I. Michalak2, N. A. Terrault3

Article first published online: 8 JUL 2010

DOI: 10.1111/j.1365-2893.2010.01321.x

© 2010 Blackwell Publishing Ltd

Issue

Journal of Viral Hepatitis

Volume 18, Issue 6, pages 415–423, June 2011

Summary.  The hepatitis B virus (HBV) replicates via an error-prone reverse

transcriptase generating potential drug-resistant quasispecies. The degree of

HBV variability in liver vs peripheral blood mononuclear cells (PBMC) in

patients on long-term suppressive antivirals is unclear. We characterized HBV

replication, drug resistance and molecular diversity in patients with plasma HBV

DNA undetectable by clinical assays. Explant liver (n = 9), PBMC (n = 6) and

plasma (n = 7) from nine such patients undergoing liver transplantation were

evaluated for HBV genomes by sensitive PCR/nucleic acid hybridization assay.

Cases with HBV DNA in liver and PBMC were tested for covalently closed circular

DNA (HBV cccDNA). HBV polymerase (P) amplicons were cloned, sequenced and both P

and overlapping surface (S) gene sequences were analysed. HBV DNA was detected

in 43% (3/7) of plasma, 100% (9/9) of liver and 83% (5/6) of PBMC samples. HBV

cccDNA was detected in all liver and one PBMC sample. Four patients had a

clinical diagnosis of resistance. HBV P gene sequencing revealed 100% wild type

(wt) in plasma (2/2), 83% wt in PBMC (5/6) but livers of 3/9 (33%) contained wt

and 6/9 (66%) carried resistance to lamivudine and/or adefovir. The translated S

gene revealed no changes affecting HBV antigenicity. Sequences from livers with

antiviral resistant mutants revealed greater interpatient quasispecies

diversity. Despite apparent HBV suppression, the liver continues to support HBV

replication and extrahepatic HBV can be detected. PBMC may be a sanctuary for wt

virus during antiviral therapy, while the liver harbours more drug-resistant

viruses. Drug resistance correlates with intrahepatic viral diversity.

[Guest guest]

http://onlinelibrary.wiley.com/doi/10.1111/j.1365-2893.2010.01321.x/abstract

Molecular characterization of intrahepatic and extrahepatic hepatitis B virus

(HBV) reservoirs in patients on suppressive antiviral therapy

C. S. Coffin1, P. M. Mulrooney-Cousins2, M. G. s3, G. van Marle4, J. P.

5, T. I. Michalak2, N. A. Terrault3

Article first published online: 8 JUL 2010

DOI: 10.1111/j.1365-2893.2010.01321.x

© 2010 Blackwell Publishing Ltd

Issue

Journal of Viral Hepatitis

Volume 18, Issue 6, pages 415–423, June 2011

Summary.  The hepatitis B virus (HBV) replicates via an error-prone reverse

transcriptase generating potential drug-resistant quasispecies. The degree of

HBV variability in liver vs peripheral blood mononuclear cells (PBMC) in

patients on long-term suppressive antivirals is unclear. We characterized HBV

replication, drug resistance and molecular diversity in patients with plasma HBV

DNA undetectable by clinical assays. Explant liver (n = 9), PBMC (n = 6) and

plasma (n = 7) from nine such patients undergoing liver transplantation were

evaluated for HBV genomes by sensitive PCR/nucleic acid hybridization assay.

Cases with HBV DNA in liver and PBMC were tested for covalently closed circular

DNA (HBV cccDNA). HBV polymerase (P) amplicons were cloned, sequenced and both P

and overlapping surface (S) gene sequences were analysed. HBV DNA was detected

in 43% (3/7) of plasma, 100% (9/9) of liver and 83% (5/6) of PBMC samples. HBV

cccDNA was detected in all liver and one PBMC sample. Four patients had a

clinical diagnosis of resistance. HBV P gene sequencing revealed 100% wild type

(wt) in plasma (2/2), 83% wt in PBMC (5/6) but livers of 3/9 (33%) contained wt

and 6/9 (66%) carried resistance to lamivudine and/or adefovir. The translated S

gene revealed no changes affecting HBV antigenicity. Sequences from livers with

antiviral resistant mutants revealed greater interpatient quasispecies

diversity. Despite apparent HBV suppression, the liver continues to support HBV

replication and extrahepatic HBV can be detected. PBMC may be a sanctuary for wt

virus during antiviral therapy, while the liver harbours more drug-resistant

viruses. Drug resistance correlates with intrahepatic viral diversity.