EASL2015:前C突变的HBV病毒性VIVO IMPACT 复制能力，病毒蛋白加工

StephenW

P0546
IN VIVO IMPACT OF PRECORE MUTATIONS OF HBV ON VIRAL
REPLICATIVE CAPACITY, VIRAL PROTEIN PROCESSING, AND
VIRION ASSEMBLY. MOLECULAR CORRELATES OF HISTOLOGIC
AND IMMUNOHISTOCHEMICAL ASPECTS IN LIVER TISSUE FROM
PATIENTS WITH CHRONIC HEPATITIS B
C. Minosse1, A. Baiocchini1, S. Coen1, P. Zaccaro1, U. Visco
Comandini1, R. Lionetti1, M. Montalbano1, F. Del Nonno1,
M. Vivarelli2, G. D’Offizi1, M.R. Capobianchi1, S. Menzo1,2. 1National
Institute for Infectious Diseases “L. Spallanzani”, Rome, 2Universit`a
Politecnica delle Marche, Ancona, Italy
E-mail: [email protected]
Background and Aims: In chronic hepatitis B (CHB), ccc- and
total intrahepatic DNA are the best markers of viral hepatic
colonization and replicative capacity. Another interesting marker
of viral colonization is histochemical staining of viral antigens.
Such staining also reveals the intracellular localization pattern
of viral antigens. In particular, HBsAg can be found either
on intracytoplasmatic membranes or on the cell membrane
in different patients. The molecular mechanisms determining
this difference, and its pathogenetic or clinical implications are
presently unknown
Methods: 35 consecutive untreated CHB patients have been
enrolled for this study. In addition to classical virological and clinical
parameters, intrahepatic cccDNA, total DNA (by real-time PCR) and
intrahepatic viral antigen distribution (by immunohistochemical
staining of HBcAg and HBsAg) were evaluated in bioptic liver tissue
from these patients. Nucleotide sequencing of the pre-core and
pre-s regions of viral genome was performed on intrahepatic and
on peripheral blood virus
Results: A strong correlation between the frequency of HBcAg
positive cells in the liver tissue and cccDNA molecules /105 cells
was found. HBcAg positive cells also correlate with viral DNA
load, HBsAg and especially HBeAg load in peripheral blood. By
contrast, intrahepatic HBsAg positive cells did not significantly
correlate with any of the different molecular parameters, but
only with peripheral HBsAg. The different patterns of intracellular
localization of HBsAg correlates significantly with viral replicative
capacity (intrahepatic totalDNA/cccDNA). More insight into this
aspect emerged by sequencing the pre-core region, which identified
pre-core mutations (but not being HBeAg negative per se) as a
strong correlate of intracytoplasmatic localization of HBsAg. In
addition, a patient with grade 3 steatosis harbored a virus with
a frameshifting deletion in the pre-core region which determined
the expression of a 51 residues long peptide replacing HBeAg
Conclusions: This study indicates that pre-core mutations in the
HBV genomes, in addition to abrogating HBeAg expression, lead to a
significant reduction of viral replicative capacity, alter intracellular
processing of viral proteins and virion assembly. Unexplained
hepatic steatosis, a rare event in the natural history of CHB, might be
a result of peculiar genetic determinants imposed by the immune
system on the virus.

StephenW

Showing translation for P0546 IN VIVO IMPACT OF PRECORE MUTATIONS OF HBV ON VIRAL REPLICATIVE CAPACITY, VIRAL PROTEIN PROCESSING, AND VIRION ASSEMBLY. MOLECULAR CORRELATES OF HISTOLOGIC AND IMMUNOHISTOCHEMICAL ASPECTS IN LIVER TISSUE FROM PATIENTS WITH CHRONIC HEPATITIS B C. Minosse1, A. Baiocchini1, S. Coen1, P. Zaccaro1, U. Visco Comandini1, R. Lionetti1, M. Montalbano1, F. Del Nonno1, M. Vivarelli2, G. D’Offizi1, M.R. Capobianchi 1, S. Menzo1,2. 1National Institute for Infectious Diseases “L. Spallanzani”, Rome, 2Universit`a Politecnica delle Marche, Ancona, Italy E-mail: [email protected] Background and Aims: In chronic hepatitis B (CHB), ccc- and total intrahepatic DNA are the best markers of viral hepatic colonization and replicative capacity. Another interesting marker of viral colonization is histochemical staining of viral antigens. Such staining also reveals the intracellular localization pattern of viral antigens. In particular, HBsAg can be found either on intracytoplasmic membranes or on the cell membrane in different patients. The molecular mechanisms determining this difference, and its pathogenetic or clinical implications are presently unknown Methods: 35 consecutive untreated CHB patients have been enrolled for this study. In addition to classical virological and clinical parameters, intrahepatic cccDNA, total DNA (by real-time PCR) and intrahepatic viral antigen distribution (by immunohistochemical staining of HBcAg and HBsAg) were evaluated in bioptic liver tissue from these patients. Nucleotide sequencing of the pre-core and pre-s regions of viral genome was performed on intrahepatic and on peripheral blood virus Results: A strong correlation between the frequency of HBcAg positive cells in the liver tissue and cccDNA molecules /105 cells was found. HBcAg positive cells also correlate with viral DNA load, HBsAg and especially HBeAg load in peripheral blood. By contrast, intrahepatic HBsAg positive cells did not significantly correlate with any of the different molecular parameters, but only with peripheral HBsAg. The different patterns of intracellular localization of HBsAg correlates significantly with viral replicative capacity (intrahepatic totalDNA/cccDNA). More insight into this aspect emerged by sequencing the pre-core region, which identified pre-core mutations (but not being HBeAg negative per se) as a strong correlate of intracytoplasmatic localization of HBsAg. In addition, a patient with grade 3 steatosis harbored a virus with a frameshifting deletion in the pre-core region which determined the expression of a 51 residues long peptide replacing HBeAg Conclusions: This study indicates that pre-core mutations in the HBV genomes, in addition to abrogating HBeAg expression, lead to a significant reduction of viral replicative capacity, alter intracellular processing of viral proteins and virion assembly. Unexplained hepatic steatosis, a rare event in the natural history of CHB, might be a result of peculiar genetic determinants imposed by the immune system on the virus.
Translate instead from P0546 IN VIVO IMPACT OF PRECORE MUTATIONS OF HBV ON VIRAL REPLICATIVE CAPACITY, VIRAL PROTEIN PROCESSING, AND VIRION ASSEMBLY. MOLECULAR CORRELATES OF HISTOLOGIC AND IMMUNOHISTOCHEMICAL ASPECTS IN LIVER TISSUE FROM PATIENTS WITH CHRONIC HEPATITIS B C. Minosse1, A. Baiocchini1, S. Coen1, P. Zaccaro1, U. Visco Comandini1, R. Lionetti1, M. Montalbano1, F. Del Nonno1, M. Vivarelli2, G. D’Offizi1, M.R. Capobianchi1, S. Menzo1,2. 1National Institute for Infectious Diseases “L. Spallanzani”, Rome, 2Universit`a Politecnica delle Marche, Ancona, Italy E-mail: [email protected] Background and Aims: In chronic hepatitis B (CHB), ccc- and total intrahepatic DNA are the best markers of viral hepatic colonization and replicative capacity. Another interesting marker of viral colonization is histochemical staining of viral antigens. Such staining also reveals the intracellular localization pattern of viral antigens. In particular, HBsAg can be found either on intracytoplasmatic membranes or on the cell membrane in different patients. The molecular mechanisms determining this difference, and its pathogenetic or clinical implications are presently unknown Methods: 35 consecutive untreated CHB patients have been enrolled for this study. In addition to classical virological and clinical parameters, intrahepatic cccDNA, total DNA (by real-time PCR) and intrahepatic viral antigen distribution (by immunohistochemical staining of HBcAg and HBsAg) were evaluated in bioptic liver tissue from these patients. Nucleotide sequencing of the pre-core and pre-s regions of viral genome was performed on intrahepatic and on peripheral blood virus Results: A strong correlation between the frequency of HBcAg positive cells in the liver tissue and cccDNA molecules /105 cells was found. HBcAg positive cells also correlate with viral DNA load, HBsAg and especially HBeAg load in peripheral blood. By contrast, intrahepatic HBsAg positive cells did not significantly correlate with any of the different molecular parameters, but only with peripheral HBsAg. The different patterns of intracellular localization of HBsAg correlates significantly with viral replicative capacity (intrahepatic totalDNA/cccDNA). More insight into this aspect emerged by sequencing the pre-core region, which identified pre-core mutations (but not being HBeAg negative per se) as a strong correlate of intracytoplasmatic localization of HBsAg. In addition, a patient with grade 3 steatosis harbored a virus with a frameshifting deletion in the pre-core region which determined the expression of a 51 residues long peptide replacing HBeAg Conclusions: This study indicates that pre-core mutations in the HBV genomes, in addition to abrogating HBeAg expression, lead to a significant reduction of viral replicative capacity, alter intracellular processing of viral proteins and virion assembly. Unexplained hepatic steatosis, a rare event in the natural history of CHB, might be a result of peculiar genetic determinants imposed by the immune system on the virus.
P0546
前C突变的HBV病毒性VIVO IMPACT
复制能力，病毒蛋白加工，
病毒粒子组装。分子组织学相关的
肝组织和免疫组织化学方面，从
与慢性乙肝患者
C. Minosse1，A. Baiocchini1，S. Coen1，P. Zaccaro1，粘U.
Comandini1，R. Lionetti1，M. Montalbano1，F.德尔Nonno1，
M. Vivarelli2，G. D'Offizi1，MR Capobianchi 1，S. Menzo1,2。 1个国家
研究所传染病“L.斯帕兰扎尼“，罗马，2Universit`a
POLITECNICA马尔凯安科纳，意大利
电子邮件：[email protected]
背景和目的：在慢性乙型肝炎（CHB），CCC-和
肝内总DNA病毒是肝脏的最好的标志
殖民化和复制能力。另一个有趣的标志
病毒定植是病毒抗原组织化学染色。
这种染色还揭示了细胞内定位模式
病毒抗原。特别是，HBsAg的既可以发现
在胞浆内的膜或在细胞膜上
在不同的病人。确定的分子机制
这种差异，并且其发病或临床意义是
目前未知
方法：35个连续未经CHB患者已
参加这项研究。除了经典病毒学和临床
参数，肝内的cccDNA，总DNA（通过实时PCR）和
肝内病毒抗原分布（免疫组化
核心抗原和乙肝表面抗原）的染色活检肝组织进行了评估
从这些患者。预芯的核苷酸测序和
病毒基因组的前S区中的肝内执行，并且
外周血病毒
结果：HBcAg的频率之间的强相关性
阳性细胞在肝组织和cccDNA的分子/ 105细胞
被发现。核心抗原阳性细胞还与相关病毒DNA
负荷，HBsAg和外周血尤其HBeAg的负载。由
相反，肝内HBsAg阳性细胞不显著
与任何不同的分子参数的相关性，但
只有与周边的HBsAg。细胞内的不同的模式
乙肝表面抗原的定位与病毒复制显著相关
容量（肝内totalDNA / cccDNA的）。更深入地了解这
方面出现通过测序前的核心区域，其中确定
前核心突变（但并非HBeAg阴性本身），为
强劲的HBsAg intracytoplasmatic本地化相关。在
此外，3级脂肪变性病人窝藏病毒与
一个移码缺失，其中所确定的前核心区
51个残基肽替代HBeAg的表达
结论：这项研究表明，在这前核心突变
乙肝病毒基因组中，除了废除HBeAg的表达，导致
显著减少病毒的复制能力，改变细胞内
处理病毒蛋白和病毒粒子组装的。原因不明
肝脂肪变性，慢性乙型肝炎的自然历史罕见的事件，可能是
由免疫强加特殊遗传决定的结果
系统上的病毒。