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Alteration of gene expression in human hepatocellular carcinoma

with integrated hepatitis B virus DNA
Tamori A, Yamanishi Y, Kawashima S, Kanehisa M, Enomoto M,
Tanaka H, Kubo S, Shiomi S, Nishiguchi S
Clin Cancer Res
11(16):5821-6, 2005
PURPOSE: Integration of hepatitis B virus (HBV) DNA into the human genome is
one of the most important steps in HBV-related carcinogenesis. This study
attempted to find the link between HBV DNA, the adjoining cellular sequence, and
altered gene expression in hepatocellular carcinoma (HCC) with integrated HBV
DNA. EXPERIMENTAL DESIGN: We examined 15 cases of HCC infected with HBV
by cassette ligation-mediated PCR. The human DNA adjacent to the integrated HBV
DNA was sequenced. Protein coding sequences were searched for in the human
sequence. In five cases with HBV DNA integration, from which good quality RNA was
extracted, gene expression was examined by cDNA microarray analysis. RESULTS:
The human DNA sequence successive to integrated HBV DNA was determined in the
15 HCCs. Eight protein-coding regions were involved: ras-responsive element
binding protein 1, calmodulin 1, mixed lineage leukemia 2 (MLL2), FLJ333655,
LOC220272, LOC255345, LOC220220, and LOC168991. The MLL2 gene was
expressed in three cases with HBV DNA integrated into exon 3 of MLL2 and in one
case with HBV DNA integrated into intron 3 of MLL2. Gene expression analysis
suggested that two HCCs with HBV integrated into MLL2 had similar patterns of
gene expression compared with three HCCs with HBV integrated into other loci of
human chromosomes. CONCLUSIONS: HBV DNA was integrated at random sites of
human DNA, and the MLL2 gene was one of the targets for integration. Our results
suggest that HBV DNA might modulate human genes near integration sites,
followed by integration site-specific expression of such genes during

Interferon-alpha treatment of patients with Hepatitis C: the role of a
comprehensive risk-benefit assessment [letter]
Rifai M A, Loftis J M, Hauser P
CNS Drugs
19(8):719-21, 2005
TT viruses: how much do we know?
Abraham P
Indian J Med Res
122(1):7-10, 2005
Presence of TT virus infection in chronic hepatitis patients from a
hospital in New Delhi, India
Chattopadhyay S, Das B C, Gupta R K, Kar P
Indian J Med Res
122(1):29-33, 2005
BACKGROUND & OBJECTIVE: The recent discovery of a novel parenterally
transmitted, unenveloped, single-stranded DNA virus called TT virus (TTV) in chronic
hepatitis patients with unclear pathogenesis throughout the world led us to
investigate, its presence in chronic hepatitis patients attending a hospital in New
Delhi, India, and to evaluate its role in liver disease. METHODS: TT virus DNA was
investigated in serum samples of 70 patients with various types of chronic hepatitis,
and 100 healthy subjects from New Delhi, India by nested PCR using the primers
that belonged to UTR (A) region of the genome. RESULTS: TTV DNA was detected
in 6 of 23 patients (26%) with type B chronic hepatitis, 3 of 20 patients (15%) with
type C chronic hepatitis, and 12 of 100 subjects (12%) from healthy control group
with normal liver function profile tests. None of the 27 non-B, non-C chronic
hepatitis patients had TTV DNA positivity. The prevalence of TTV was significantly

higher in type-B chronic hepatitis patients as compared to normal subjects (P< 0.05)
but comparable to type C chronic hepatitis patients. The clinical course and
biochemical profiles of type B, or type C chronic hepatitis patients co-infected with
TTV did not differ significantly from those without TTV infection. INTERPRETATION
& CONCLUSION: Interestingly, in chronic hepatitis patients, TTV was always
associated with either hepatitis B or C virus indicating a likely parenteral route of
transmission. All TTV-positive subjects in healthy control group showed normal
clinical and biochemical profiles. Thus, the presence of TTV infection is unlikely to
influence the course of chronic hepatitis related to hepatitis B virus (HBV) or
hepatitis C virus (HCV) or cause liver diseases in healthy subjects.

Cirrhosis in hepatitis C virus-infected patients can be excluded using
an index of standard biochemical serum markers
Islam S, Antonsson L, Westin J, Lagging M
Scand J Gastroenterol
40(7):867-72, 2005
OBJECTIVE: Assessment of liver histology is pivotal in prognostication and
decision-making regarding therapeutic intervention in patients with chronic
hepatitis C virus (HCV). Being an invasive procedure, the liver biopsy is associated
with complications, and a non-invasive alternative would be preferable. MATERIAL
AND METHODS: Sera samples from 179 patients with chronic HCV infection
collected at the time of liver biopsy were analyzed using routinely available
biochemical markers of liver disease, and liver histology was evaluated using the
Ishak protocol. The relationship between the serum biochemical markers and
cirrhosis (Ishak stage > or = 5) as well as bridging fibrosis (Ishak stage > or = 3)
was examined. RESULTS: A strong association was found in the multivariate
logistic regression analysis between fibrosis stage and aspartate aminotransferase
(AST), platelet count and prothrombin-INR (international normalized ratio). An index
(the Goteborg University Cirrhosis Index (GUCI)) was calculated using these
variables: normalized ASTxprothrombin-INRx100/platelet count (x 10(9)/l). Using a
cut-off value of 1.0, the sensitivity was 80% and the specificity 78% for diagnosis of
cirrhosis, and the negative predictive values (NPV) and positive predictive values
(PPV) were 97% and 31%, respectively. The GUCI score proved slightly superior for
sensitivity, specificity, NPV, PPV, and the area under the receiver operating
characteristic (ROC) curve for prediction of cirrhosis and bridging fibrosis compared
with the AST to platelet ratio index (APRI), which has been reported as a predictor of
significant fibrosis and cirrhosis. CONCLUSION: An index using routinely available
biochemical markers can with a high degree of accuracy discriminate patients with
from those without hepatitis C-related cirrhosis.

Acute hepatitis E infection in a visitor to Louisiana
Walker J E, Brown T M, Hagood T L, Cassidy W M
South Med J
98(7):721-2, 2005
A man from Africa had been visiting Mississippi and Louisiana when he had
development of acute hepatitis. Although hepatitis E is endemic to many parts of
the world and has been associated with large outbreaks, it has remained relatively
uncommon in this country. With growing foreign travel and an ever-increasing
number of cases reported nationally, hepatitis E infection should be considered
more frequently in the differential diagnosis of acute hepatitis.

Reversible encephalopathy secondary to paratyphoid infection and
concomitant acute Hepatitis A

El-Khoury M, Naoushi H, Sawaya R, Aoun E, Nassar N T, Sharara A I

South Med J
98(7):723-5, 2005
Reversible encephalopathy has been described in association with typhoid fever as
well as nontyphoidal salmonella infections. A diagnostic dilemma as to the cause of
encephalopathy may arise when there is coexistent acute viral hepatitis and
suspicion of fulminant liver failure. The authors report a patient who presented with
acute icteric hepatitis A infection and a concomitant febrile illness due to
Salmonella paratyphi associated with progressive encephalopathy and coma. The
young man developed high-grade fever and coma. Concomitant infection with S.
paratyphi was diagnosed and the patient's encephalopathy resolved rapidly with
antibiotic therapy. This is the first report of a putative association of S. paratyphi
infection and reversible encephalopathy. Salmonella and hepatitis A virus infection
are both transmitted via the fecal-oral route, and awareness of this association is
important in the management of such patients.

Breaking the chain: universal childhood hepatitis A virus vaccination
Temte J L
Am Fam Physician
73(12):2127-8, 2006
The analytic sensitivity and mutant detection capability of six
hepatitis B surface antigen assays
La'ulu S L, Roberts W L
Am J Clin Pathol.
125(5):748-51, 2006
Hepatitis B virus surface antigen (HBsAg) mutants occur in clinical specimens. We
studied the analytic sensitivity and ability to detect recombinant and native
mutants of 6 HBsAg assays. The ARCHITECT, AUSZYME MONOCLONAL and AxSYM
assays (Abbott Diagnostics, Abbott Park, IL), the ADVIA Centaur assay (Bayer
Diagnostics, Tarrytown, NY), and the Test System 3 and VITROS ECi assays (Ortho
Clinical Diagnostics, Raritan, NJ) showed comparable sensitivity with wild-type
HBsAg. The ARCHITECT, AUSZYME, and AxSYM assays detected all mutants that
were tested. The Test System 3 and VITROS ECi assays failed to detect mutants with
amino acid substitutions at positions 143, 144, and 145, which are located in the
immunodominant "a" determinant. The ADVIA Centaur failed to detect substitutions
at position 145 and showed negative or very low positive results for substitutions at
position 143. The inability to detect HBsAg mutants may lead to misdiagnosis of
hepatitis B virus infection. Further studies on the prevalence of HBsAg mutants and
the ability of commercial assays to detect them are needed.

Expression of DC-SIGN and DC-SIGNR on human sinusoidal
endothelium: a role for capturing hepatitis C virus particles
Lai W K, Sun P J, Zhang J, Jennings A, Lalor P F, Hubscher S,
McKeating J A, Adams D H
Am J Pathol
169(1):200-8, 2006
Hepatic sinusoidal endothelial cells are unique among endothelial cells in their
ability to internalize and process a diverse range of antigens. DC-SIGNR, a type 2 C-

type lectin expressed on liver sinusoids, has been shown to bind with high affinity to
hepatitis C virus (HCV) E2 glycoprotein. DC-SIGN is a closely related homologue
reported to be expressed only on dendritic cells and a subset of macrophages and
has similar binding affinity to HCV E2 glycoprotein. These receptors function as
adhesion and antigen presentation molecules. We report distinct patterns of DCSIGNR and DC-SIGN expression in human liver tissue and show for the first time that
both C-type lectins are expressed on sinusoidal endothelial cells. We confirmed that
these receptors are functional by demonstrating their ability to bind HCV E2
glycoproteins. Although these lectins on primary sinusoidal cells support HCV E2
binding, they are unable to support HCV entry. These data support a model where
DC-SIGN and DC-SIGNR on sinusoidal endothelium provide a mechanism for high
affinity binding of circulating HCV within the liver sinusoids allowing subsequent
transfer of the virus to underlying hepatocytes, in a manner analogous to DC-SIGN
presentation of human immunodeficiency virus on dendritic cells.

Separate origins of hepatitis B virus surface antigen-negative foci and
hepatocellular carcinomas in transgenic HBsAg (alb/psx) mice
Crawford D R, Ostrowski S, Vakharia D, Ilic Z, Sell S
Am J Pathol
169(1):223-32, 2006
We have examined the development and transgene expression in liver lesions of
transgenic mice bearing the hepatitis B surface antigen (HBsAg) gene of hepatitis B
virus under the control of the albumin promoter (alb/psx) to study liver regeneration
and hepatocellular carcinoma (HCC) associated with hepatitis B virus infection.
Storage of the HBsAg in the endoplasmic reticulum precedes loss of liver cells and
regenerative hyperplastic nodules that do not express HBsAg. Histological analysis
indicated that HBsAg-negative foci and nodules arose from liver progenitor cells in
the portal zone and lacked mRNA expression. Genomic DNA from eight of nine
HBsAg-negative laser capture-excised liver foci showed loss of part of the alb/psx
gene, whereas no loss of the actin gene was observed. The alb/psx DNA was intact
in adjacent HBsAg-positive tissue. Sequencing of polymerase chain reaction
products suggested that alterations in the HBsAg transgene in HBsAg-negative foci
occurred via large-scale deletions as opposed to single-site mutations. Southern blot
analysis of HCC from 2-year-old transgenic HBsAg mice, however, revealed an intact
alb/psx gene. Thus, HBsAg-negative progenitor cells with deletions in the transgene
appear to be responsible for compensatory regeneration of the liver, whereas HCCs
arise from clonal expansion of hepatocytes with intact alb/psx transgenes.

Interferon-based combination anti-viral therapy for hepatitis C virus
after liver transplantation: a review and quantitative analysis
Wang C S, Ko H H, Yoshida E M, Marra C A, Richardson K
Am J Transplant
6(7):1586-99, 2006
Recurrence of hepatitis C virus (HCV) infection after liver transplantation (LT) is
universal. However, the efficacy, tolerability and safety of combination interferon
and ribavirin (IFN-RIB) or peginterferon and ribavirin (PEG-RIB) anti-viral therapies
post-LT are uncertain. We performed a comprehensive search of major medical
databases (1980-2005) and conference proceedings (1996-2005). The main
outcome measure was sustained virological response (SVR, undetectable HCV RNA)
at 6 months. Summary estimates were calculated using random-effects models.
Twenty-seven IFN-RIB and 21 PEG-RIB studies were included. IFN-RIB was associated
with a pooled SVR rate of 24% (95% CI, 20-27%), while PEG-RIB was associated with

an SVR rate of 27% (23-31%). Pooled discontinuation rates were 24% (21-27%) with
IFN-RIB and 26% (20-32%) with PEG-RIB. The pooled rate of acute graft rejection
was 2% (1-3%) with IFN-RIB and 5% (3-7%) with PEG-RIB. IFN-RIB and PEG-RIB
therapies in HCV infection post-LT were associated with similar but overall low SVR
and were poorly tolerated. The rate of acute rejection was small. The therapeutic
advantage of PEG-RIB therapy observed in non-transplant chronic HCV infection
appears to be attenuated post-LT. Clinical trials are needed to evaluate reasons for
this post-transplant therapeutic disadvantage and to find strategies to ameliorate

Natural history of patients with recurrent chronic hepatitis C virus and
occult hepatitis B co-infection after liver transplantation
Hui C K, Lau E, Monto A, Kim M, Luk J M, Poon R T, Leung N, Lo C M,
Fan S T, Lau G K, Wright T L
Am J Transplant
6(7):1600-8, 2006
It is uncertain whether occult hepatitis B virus co-infection will hasten progressive
liver disease in chronic hepatitis C patients after liver transplantation. This study
evaluated fibrosis progression and severe fibrosis in 118 consecutive hepatitis B
surface antigen-negative patients with virological and histological evidence of
recurrent chronic hepatitis C infection co-infected with occult hepatitis B virus after
liver transplantation. HBV DNA was detected from serum at the time of recurrent
chronic hepatitis C infection by polymerase chain reaction. Each subject underwent
a repeat liver biopsy 5 years post-liver transplantation. Occult hepatitis B virus coinfection was present in 41 of the 118 (34.7%) patients. At 5 years post-liver
transplantation, 13 of the 41 occult hepatitis B virus co-infected patients compared
with 16 of the 77 patients without occult hepatitis B virus co-infection developed
fibrosis progression (31.7% vs. 20.8%, respectively, p = 0.39). Eight of 41 the occult
hepatitis B virus co-infected patients compared with 13 of the 77 patients without
occult hepatitis B virus co-infection had severe fibrosis (19.5% vs. 16.9%,
respectively, p = 0.97). In conclusion, occult hepatitis B virus co-infection in
patients with recurrent chronic hepatitis C infection was not associated with
accelerated fibrosis progression or severe fibrosis after liver transplantation.

Acute renal allograft rejection following pegylated IFN-alpha
treatment for chronic HCV in a repeat allograft recipient on
hemodialysis: a case report
Carbognin S J, Solomon N M, Yeo F E, Swanson S J, Bohen E M, Koff J
M, Sabnis S G, Abbott K C
Am J Transplant
6(7):1746-51, 2006
Interferon alpha (IFN-alpha) can be effective therapy for patients with chronic
kidney disease who have chronic hepatitis C (HCV). However, acute allograft
rejection has been reported in association with IFN-alpha following kidney
transplantation, and therefore IFN therapy is recommended prior to, rather than
after, kidney transplantation whenever feasible. The special case of repeat allograft
recipients who contract HCV after the first transplantation presents special
difficulties. This report features the case of a repeat allograft recipient who
presented with neutropenic fevers after 5 months of pegylated IFN-alpha therapy,
initiated 6 months following the functional loss of his third graft and the reinitiation
of hemodialysis (HD). Physical exam, radiographic and laboratory findings led to

allograft nephrectomy. The pathologic findings supported a diagnosis of acute-onchronic rejection. This represents a rare case of IFN-alpha induced rejection
following allograft failure and return to HD in a repeat allograft recipient. It also calls
attention to the need for a high index of suspicion for the development of allograft
rejection, which may require allograft nephrectomy even after allograft 'failure'.

Hepatitis C: reviewing the options [letter]
Khan S, Sewell W A
175(1):63, 2006
Hepatitis C: reviewing the options [letter]
Pijak M R
175(1):63-4, 2006
Hepatitis C: reviewing the options - Reply [letter]
Lee S S, Wong T
175(1):64, 2006
Dideoxynucleoside analogues should be used cautiously in patients
with hepatic steatosis [editorial]
Zeremski M, Talal A H
Clin Infect Dis
43(3):373-6, 2006
Non-invasive biomarkers of liver fibrosis in haemophilia patients with
hepatitis C: can you avoid liver biopsy?
Maor Y, Bashari D, Kenet G, Lubetsky A, Luboshitz J, Schapiro J M,
Penaranda G, Bar-Meir S, Martinowitz U, Halfon P
12(4):372-9, 2006
Introduction: Liver biopsy remains the gold standard for the evaluation of fibrosis
despite its risks and limitations, especially in haemophilia patients. Recently, noninvasive biomarkers have been used to assess histological features. The most
thoroughly evaluated biomarker is the FibroTest (FT) (AUROC 0.80 for fibrosis stages
F2F3F4 vs. F0F1). Aim: To estimate liver fibrosis in haemophilia patients infected
with hepatitis C (HCV) using non-invasive biomarkers without liver biopsy. Methods:
One hundred and thirty-two haemophilia patients (124 male, mean age 38 +/- 14
years) with anti-HCV antibodies were evaluated. These patients were stratified into
several groups: patients with features of advanced liver disease - seven,
persistently HCV RNA-negative - 21, persistently normal liver function tests (LFTs)24, HCV/HIV co-infected - 27. The following biomarkers of fibrosis were used: FT,
AST-to-platelet ratio index (APRI), Forns index, age-platelet index and hyaluronic
acid. The obtained scores were correlated with the clinical features of the patients.
Results: Estimated by the FT, the distribution of the stage of fibrosis in the 132
patients was F0F1 = 65% (86/132), F2 = 5% (7/132), F3 = 13% (17/132) and F4 =
17% (22/132). Using FT, all patients with clinical suspicion of advanced liver disease
were classified as F3F4, whereas patients with persistently HCV RNA-negative were
all classified as F0F1. Twenty-one per cent (5/24) of the patients with persistently
normal LFTs had fibrosis stage F3F4. The proportion of F3F4 among HCV/HIV coinfected patients was significantly higher than among HCV mono-infected (52% vs.
33%; P = 0.05). Concordance of three or more biomarkers was present in 43%

(57/132) of the patients. Liver biopsy could be avoided in 70% (92/132) using a
practical assumption that if FT is in concordance with APRI and/or Forns, then we
may confidently rely on the biomarker. Concordance rate for patients with
presumably advanced or minimal liver disease was excellent (100% and 95%
respectively). Conclusions: In our HCV-infected haemophilia patients, FT correctly
identified clinically advanced or minimal liver disease. Discordance among the
various biomarkers of fibrosis was considerate; nevertheless, practical combination
of FT, APRI, and Forns may predict stage of fibrosis with accuracy, potentially
avoiding liver biopsy in the majority of the patients.

Preliminary results of HBV DNA testing of Polish haemophilia patients
- lack of occult HBV infection
Windyga J, Brojer E, Gronowska A, Grabarczyk P, Mikulska K,
Szczepanik A B, Stefanska E, Buczma A
12(4):380-3, 2006
Identification of hepatitis B virus (HBV) infection in the absence of surface antigen
(HbsAg) became possible with the introduction of HBV DNA detection methods.
Such occult HBV infection was diagnosed recently in about half of the Japanese
HBsAg-negative haemophilia patients. The aim of our study was to assess the
prevalence of occult HBV infection in Polish severe haemophilia population on the
sample of 115 haemophilia A and B patients (mean age 34.9 +/- 10.9) treated with
non-virus inactivated clotting factor preparations before 1995. HBV DNA was
detected in nine HBsAg-positive patients (7.8%). The mean HBV DNA load was 72
800 IU mL(-1) (250-400 000 IU mL(-1)). Hepatitis C virus (HCV) RNA was found in six
out of nine HBV-positive patients. In conclusion, HBV DNA was identified only in
HBsAg-positive patients. Unlike in Japan, the frequency of occult HBV infection in
Polish haemophilia population seems extremely rare or absent.

Should aged patients with chronic hepatitis C be treated with
interferon and ribavirin combination therapy?
Hiramatsu N, Oze T, Tsuda N, Kurashige N, Koga K, Toyama T,
Yasumaru M, Kanto T, Takehara T, Kasahara A, Kato M, Yoshihara
H, Katayama K, Hijioka T, Hagiwara H, Kubota S, Oshita M, Haruna
Y, Mita E, Suzuki K, Ishibashi K, Hayashi N
Hepatol Res
35(3):185-9, 2006
The aim of this study was to investigate the efficacy and safety of combination
therapy of interferon and ribavirin for aged patients with chronic hepatitis C.
METHODS: This study was conducted at Osaka University Hospital and institutions
participating in the Osaka Liver Disease Study Group on 329 patients with chronic
hepatitis C receiving interferon and ribavirin combination therapy (group A, under
60 year old, n=199; group B, 60-64 year old, n=64; group C, over 65 year old (mean
age, 67.8+/-2.2 year old, n=66)). Of the 293 patients who were tested for HCV
serotype and HCV viral loads, 215 had HCV-RNA with serotype 1 and high viral loads
(1H) and the other 78 had HCV-RNA with serotype 2 or low viral loads (non-1H).
RESULTS: In per-protocol analysis, the overall SVR rate of 1H patients was 28%
(51/184). Among the 1H patients, the SVR rate was significantly lower in group C
(16%) and group B (17%) than in group A (34%) (p<0.05). The overall SVR rate of
non-1H patients was 85% (57/67). No significant difference was found in the SVR
rate among group C (79%), group B (100%), and group A (84%). On the other hand,
the discontinuance of both drugs due to side effects was 29% (19/66) in group C,
20% (13/64) in group B, and 11% (21/199) in group A, with the discontinuance rates
being higher in the older group (p=0.002). CONCLUSIONS: In aged chronic hepatitis
C patients, interferon and ribavirin combination therapy can be recommended for
the non-1H patients who showed a high SVR rate of approximately 65%, but not for
the 1H patients.

Hepatitis C virus genotype and HIV coinfection affect cytokine mRNA
levels in unstimulated PBMC but do not shift the T1/T2 balance
Lee S, Watson M W, Clark B, Flexman J P, Cheng W, French M A,
Price P
Immunol Cell Biol
84(4):390-5, 2006
Rapid progression of hepatitis C virus (HCV) disease in patients with HIV/HCV may
reflect different cytokine responses and be influenced by HCV genotype. This is
addressed by a study of patients with HIV/HCV coinfection and infection with HCV
genotype 2 or 3 (2/3). They are compared with coinfected patients infected with
genotype 1 and HCV monoinfected patients matched for HCV genotype. IFNgamma, IL-10, IL-4 and IL-4delta2 mRNA were quantified by real-time PCR in
unstimulated PBMC and after in vitro stimulation with HCV core or nonstructural
3/4A antigen. In unstimulated PBMC, levels of IFN-gamma and IL-4 mRNA were
lowest in HIV/HCV genotype 1 patients, intermediate in HIV/HCV genotype 2/3
patients and highest in HCV genotype 2/3 patients. Neither HCV genotype nor HIV
affected levels of IL-10 mRNA in unstimulated PBMC or IFN-gamma, IL-4 and IL-10
mRNA in PBMC stimulated with HCV antigens. Levels of IL-4 and IL-4delta2 mRNA
correlated in mitogen-stimulated PBMC from all patient groups but both were low in
HIV/HCV genotype 1 patients. Serum soluble CD30 levels (a putative marker of a T2
cytokine environment) did not differ between patient groups. The data do not
suggest a shift in the T1/T2 balance driven by HIV coinfection or HCV genotype but
either may affect IL-4 bioavailability.

Hepatitis B - a review of the role of NAT in enhancing blood safety
Lelie N, Heaton A
J Clin Virol
36(Suppl 1):S1-2, 2006
Genetic variation in HBV infection: genotypes and mutants
Gunther S
J Clin Virol
36(Suppl 1):S3-11, 2006
Epidemiology of Hepatitis B virus and genotype
Allain J P
J Clin Virol
36(Suppl 1):S12-7, 2006
Breakthrough of hepatitis B virus escape mutants after vaccination
and virus reactivation
Gerlich W H
J Clin Virol
36(Suppl 1):S18-22, 2006
Escape mutants of hepatitis B virus (HBV) carry mutations in the major antigenic
region of the hepatitis B surface antigen (HBsAg). They are able to grow in the
presence of antibodies against HBsAg (anti-HBs) and may escape detection by
immuno assays for HBsAg. Antibodies against HBV core antigen (anti-HBc) are
considered to be a universal marker for active or resolved HBV infections but they

may appear very late. Highly sensitive detection of HBV DNA is the most universal
reliable marker of infectivity, but in the blood donation setting even the most
sensitive assay for HBV DNA may not detect all infectious donations. This brief
report describes three cases of HBV infections caused by HBV escape mutants. The
implications of the findings for the prevention of HBV transmission through donor
blood are discussed.

Assessing the impact of HBV NAT on window period reduction and
residual risk
Kleinman S H, Busch M P
J Clin Virol
36(Suppl 1):S23-9, 2006

Blood screening for HBV DNA
Fang C T
J Clin Virol
36(Suppl 1):S30-2, 2006
Epidemiology of HBV infection in Asian blood donors: emphasis on
occult HBV infection and the role of NAT
Liu C J, Chen D S, Chen P J
J Clin Virol
36(Suppl 1):S33-44, 2006
Hepatitis B virus (HBV) infection is endemic in many Asian countries. Among many
transmission routes, transfusion is the one that should be prevented. The first major
success in enhancing transfusion safety came with the implementation of hepatitis
B surface antigen (HBsAg) in the early 1970s. However, the studies quoted in this
review demonstrate that transmission by blood components negative for HBsAg can
still occur in the acute phase of infection during the seronegative window period, or
during chronic stages of infection (i.c. "occult" HBV infection, OHB). OHB is defined
as the presence of HBV DNA in blood or liver tissues in patients negative for HBsAg,
with or without any HBV antibodies. Because of limitations in current blood
screening practices, OHB is an overlooked source of HBV transmission. For policy
development on screening for HBV infection in blood donors, it would be useful to
assess the relative contribution of the above two sources of transfusion-transmitted
HBV infection from HBsAg-negative donations. New screening policy should be
evaluated on the basis of available data or newly designed studies. While anti-HBc
screening can climinate residual risk of occult HBV transmission by transfusion in
low-endemic areas, it would not be practical in most parts of the world where the
prevalence of anti-HBc is >10% as too many otherwise healthy donors will be
ineligible. On the contrary, studies mentioned in this paper indicate that nucleic acid
amplification test (NAT) or new HBsAg tests of enhanced sensitivity would be
effective in the screening of blood donors for OHB in highly endemic countries.
However, the cost-effectiveness of blood screening tests is a major concern in Asia.
We therefore have systemically reviewed the literature on prevalence and infectivity
of OHB in Asian countries and the possible role of NAT for identifying blood donors in
the pre-HBsAg window phase or in later stages of OHB.

Impact of hepatitis B vaccination on hepatitis B disease and nucleic
acid testing in high-prevalence populations
Chang M H

J Clin Virol

36(Suppl 1):S45-50, 2006

Hepatitis B virus (HBV) infection is highly prevalent in Asia, Africa, southern Europe
and Latin America, HBV vaccination has effectively reduced the acute and chronic
infection rates as well as related complications in the vaccinated children. The
incidence of hepatocellular carcinoma in children has been reduced to
approximately 25% of the incidence before the vaccination program, and fulminant
hepatitis in children has also been reduced after universal hepatitis B vaccination.
HBV DNA sero-positive rate was 98-100% in HBsAg positive vaccinated children,
while the positive rate was only 11-20% in those vaccinees with a negative HBsAg
but positive anti-HBc reaction. Hepatitis B surface gene mutants in HBV DNA
positive children increased gradually from 7.8% before the vaccination program, to
19.6%, 28.1% and 23.1% at 5, 10 and 15 years after the vaccination program. Longterm follow-up of vaccinated children has confirmed that universal HBV vaccination
in infancy has produced adequate protection up to 14 years of age. The annual
decay rate of hepatitis B surface antibody (anti-HBs) was 10.2% in children who did
not receive a booster dose. The new HBV infection rate was not different between
those who did and those who did not receive a booster dose of HBV vaccine. During
a follow-up period of seven years for 1200 vaccinated 7-year-old children in Taiwan,
the mean annual hepatitis B core antibody sero-conversion rate was 0.2%. All were
negative for HBV DNA. No new chronic HBV infections developed. A booster dose of
HBV vaccine is not recommended in children under 15 years of age. Systematic HBV
DNA screening of a large population such as blood donors may be instrumental in
following the long-term effect of the universal vaccination program on the incidence
of silent HBV infection and vaccine escape mutants.

Changing patterns of hepatitis B infection in Italy and NAT testing for
improving the safety of blood supply
Zanetti A R, Romano L, Zappa A, Velati C
J Clin Virol
36(Suppl 1):S51-5, 2006
In Italy, as in most industrialized countries, the burden of hepatitis B has
progressively declined in recent decades as a consequence of general
improvements in hygiene and standard of living, the introduction of several public
health measures, refinement in blood screening and the implementation of specific
vaccination programmes. Universal hepatitis B vaccination for all infants and
adolescents as well as individuals at increased risk has resulted in considerable
progress towards prevention and control of HBV infection. The residual risk of
transmitting blood-borne viruses through transfusion is currently very low. Nucleic
acid testing can shorten the window period and, consequently, further reduce the
risk of viral transmission. Blood donor screening for HCV by NAT was initiated in Italy
in 2001 and became mandatory in June 2002. NAT for HIV is currently mandatory in
four regions, not mandatory but almost universally performed in another thirteen
regions, and not yet introduced in the remaining four regions. NAT for HBV is
currently mandatory in four regions and under evaluation in the remaining. NAT for
HBV may be a useful tool in detecting acute viral infections in the window phase as
well as the occult infections. Its efficacy in improving the safety of blood supply is
expected to be higher in countries with intermediate/high endemicity, where antiHBc antibody screening cannot be routinely performed. There is agreement that, at
present, the implementation of HBV DNA testing will not allow for discontinuation of
screening for HBsAg.

Hepatitis B virus (HBV) screening strategy to ensure the safety of
blood for transfusion through a combination of immunological testing
and nucleic acid amplification testing - Japanese experience
Yugi H, Mizui M, Tanaka J, Yoshizawa H
J Clin Virol
36(Suppl 1):S56-64, 2006

Treatment of chronic hepatitis B in children
Jonas M M
J Pediatr Gastroenterol Nutr
43(1 Suppl):S56-60, 2006
Chronic hepatitis B infection remains a significant health problem worldwide, and
acquisition during infancy or childhood causes many of the chronic infections that
are responsible for the morbidity associated with this disease. Some children with
chronic hepatitis B are candidates for treatment. Two medications are currently
licensed for use in children, and it is anticipated that others will be available in the
next several years. This article describes rationale for treatment, patient selection
and pros and cons of the current and expected therapeutic options.

Development of hepatitis B oral vaccine using B-cell epitope loaded
PLG microparticles
Rajkannan R, Dhanaraju M D, Gopinath D, Selvaraj D, Jayakumar R
24(24):5149-57, 2006
Oral hepatitis B vaccine formulation was prepared by successful encapsulation of
immunogenic peptide representing residues 127-145 of the immunodominant B-cell
epitope of hepatitis B surface antigen (HBsAg) in poly(d,l-lactide co-glycolide) (PLG)
microparticles. The smooth, spherical PLG microparticles with a diameter of around
10mum was prepared by using W/O/W double emulsion solvent evaporation
method. The entrapment efficiency of B-cell epitope peptide (BCEP) into PLG
microparticles was 64%. In vitro studies showed B-cell epitope loaded PLG
microparticles (BCEM) released the peptide in sustained profile and reached 64.9%
efficiency by Day 25. Single oral immunization of mice with BCEM led to the
significant induction of specific serum IgG and IgM anti-HB antibodies. After the
termination of antibody induction, the orally immunized mice were infected with
HBsAg, which resulted in the rapid production of antibodies against HBsAg as a
result of secondary immune response. PLG microparticles formulation approach may
have potential in increasing the efficacy of microparticulate systems for the oral
administration of hepatitis B vaccine.

DNA vaccines expressing the duck hepatitis B virus surface proteins
lead to reduced numbers of infected hepatocytes and protect ducks
against the development of chronic infection in a virus dosedependent manner
Miller D S, Kotlarski I, Jilbert A R
351(1):159-69, 2006
We tested the efficacy of DNA vaccines expressing the duck hepatitis B virus (DHBV)
pre-surface (pre-S/S) and surface (S) proteins in modifying the outcome of infection
in 14-day-old ducks. In two experiments, Pekin Aylesbury ducks were vaccinated on
days 4 and 14 of age with plasmid DNA vaccines expressing either the DHBV pre-S/S
or S proteins, or the control plasmid vector, pcDNA1.1Amp. All ducks were then
challenged intravenously on day 14 of age with 5 x 10(7) or 5 x 10(8) DHBV
genomes. Levels of initial DHBV infection were assessed using liver biopsy tissue
collected at day 4 post-challenge (p.c.) followed and immunostained for DHBV
surface antigen to determine the percentage of infected hepatocytes. All vector
vaccinated ducks challenged with 5 x 10(7) and 5 x 10(8) DHBV genomes had an
average of 3.21% and 20.1% of DHBV-positive hepatocytes respectively at day 4


p.c. and 16 out of 16 ducks developed chronic DHBV infection. In contrast, pre-S/S
and S vaccinated ducks challenged with 5 x 10(7) DHBV genomes had reduced
levels of initial infection with an average of 1.38% and 1.93% of DHBV-positive
hepatocytes at day 4 p.c. respectively and 10 of 18 ducks were protected against
chronic infection. The pre-S/S and the S DNA vaccinated ducks challenged with 5 x
10(8) DHBV genomes had an average of 31.5% and 9.2% of DHBV-positive
hepatocytes on day 4 p.c. respectively and only 4 of the 18 vaccinated ducks were
protected against chronic infection. There was no statistically significant difference
in the efficacy of the DHBV pre-S/S or S DNA vaccines. In conclusion, vaccination of
young ducks with DNA vaccines expressing the DHBV pre-S/S and S proteins
induced rapid immune responses that reduced the extent of initial DHBV infection in
the liver and prevented the development of chronic infection in a virus dosedependent manner.

Pegylated-interferon alpha 2a treatment for chronic hepatitis C in
patients on chronic haemodialysis
Sporea I, Popescu A, Sirli R, Golea O, Totolici C, Danila M, Vernic C
World J Gastroenterol
12(26):4191-4, 2006
AIM: To evaluate the response to pegylated-interferon alpha 2a in chronic hepatitis
C patients on chronic haemodialysis. METHODS: Ten patients with chronic C
hepatitis were enrolled in this study. All had increased aminotransferases for more
than 6 mo, positive antiHCV antibodies and positive PCR HCV-RNA. We
administrated Peg-Interferon alpha 2a 180 mug/wk for 48 wk. After 12 wk of
treatment we evaluated the biochemical and early virological response (EVR). At the
end of the treatment we evaluated the biochemical response and 24 wk after the
end of the treatment we evaluated the sustained virological response (SVR). We
monitored the side-effects during the treatment. RESULTS: Two patients dropped
out in the first 12 wk of treatment and 2 after the first 12 wk of treatment. After 12
wk of treatment, 7 out of 8 patients had biochemical response and EVR and 1 had
biochemical response but persistent viremia. We had to reduce the dose of
pegylated-interferon to 135 mug/wk in 2 cases. Three out of 6 (50%) patients had
SVR 24 wk after the end of the treatment. Intention-to-treat analysis showed that 3
out of 10 patients (30%) had SVR. Side-effects occurred in most of the patients (flulike syndrome, thrombocytopenia or leucopoenia), but they did not impose the
discontinuation of treatment. CONCLUSION: After 12 wk of treatment with PegInterferon alpha 2a (40 ku) in patients on chronic haemodialysis with chronic C
hepatitis, EVR was obtained in 87.5% (7/8) of the cases. SVR was achieved in 50%
of the cases (3/6 patients) that finished the 48 wk of treatment.

Biochemical response to lamivudine treatment in HBeAg negative
chronic hepatitis B patients in Iran
Mohammad Alizadeh A H, Ranjbar M, Karimi B, Hatami S
World J Gastroenterol
12(26):4203-5, 2006
AIM: To study the effect of a one-year lamivudine regimen in patients with chronic
hepatitis B. METHODS: Medical records of HBeAg negative hepatitis B patients who
attended a hepatitis clinic in Tehran between March 2002-March 2004 were
evaluated. The patients received 100 mg lamivudine tablets once daily for at least
12 mo. Liver enzymes and complete blood count were checked at baseline and the
end of treatment (12th mo) and 6 mo after discontinuation of treatment. RESULTS:
Of all patients, 24 were excluded. Of 71 patients left, 58 (81.7%) were men. Mean
age of the patients was 38 +/- 14 years. Mean level of ALT in serum was 1437 +/205 nkat/L at baseline with a significant reduction at the end of treatment to a


mean level of 723 +/- 92 nkat/L (P = 0.002). In 38 patients (53.5%), the ALT level
was normal after one-year treatment. Five patients (7.3%) relapsed (biochemically)
within 6 mo after discontinuing lamivudine therapy (the patients with good end of
treatment response). Mean level of AST in serum was 1060 +/- 105 nkat/L at
baseline which decreased significantly to 652 +/- 75 nkat/L at the end of treatment
(P = 0.002). CONCLUSION: Over half (53.5%) of chronic hepatitis B patients with
HBeAg negative have normal liver enzyme level at 12-mo lamivudine therapy.

Mutations in surface and polymerase gene of chronic hepatitis B
patients with coexisting HBsAg and anti-HBs
Lu H Y, Zeng Z, Xu X Y, Zhang N L, Yu M, Gong W B
World J Gastroenterol
12(26):4219-23, 2006
AIM: To investigate the clinical significance and presence of mutations in the
surface (S) and overlapping polymerase gene of hepatitis B patients with coexisting
HBsAg and anti-HBs. METHODS: Twenty-three patients with chronic hepatitis B
were studied. Of the 23 patients, 11 were both positive for hepatitis B virus (HBV)
surface antigen (HBsAg) and antibody to HBV surface antigen (anti-HBs), 12 were
negative for anti-HBs while positive for HBsAg. DNA was extracted from 200 muL
serum of the patients. Nucleotide of the surface and overlapping polymerase gene
from HBV-infected patients was amplified by PCR, and the PCR products were
sequenced. RESULTS: Forty-one mutations were found within the surface gene
protein of HBV in 15 patients (10 with coexisting HBsAg and anti-HBs). Six (14.6%)
out of 41 mutations were located at "alpha " determinant region in 5 patients (4
positive for HBsAg and anti-HBs). Eleven mutations (26.8%) occurred in the
downstream or upstream of "alpha " determinant region. Lamivudine (LMV)-selected
mutations were found in three patients who developed anti-HBs, which occurred in
amino acid positions (196, 198, 199) of the surface protein and in YMDD motif
(M204I/V) of the polymerase protein simultaneously. Presence of these mutations
did not relate to changes in ALT and HBV DNA levels. CONCLUSION: Besides
mutations in the "alpha " deter-minant region, mutations at downstream or
upstream of the "alpha " determinant region may contribute to the development of
anti-HBs. These mutations do not block the replicating competency of HBV in the
presence of high titer of anti-HBs.

Investigation on correlation between expression of CD58 molecule
and severity of hepatitis B
Sheng L, Li J, Qi B T, Ji Y Q, Meng Z J, Xie M
World J Gastroenterol
12(26):4237-40, 2006

AIM: To investigate the correlation between expression of CD58 and severity of

hepatitis B. METHODS: The level of soluble CD58 (sCD58) in serum of patients with
hepatitis B was detected by enzyme-linked immunosorbent assay. The level of
expression of membrane CD58 molecule in PBMC was detected by direct
immunofluorescence. The levels of serumal TBIL, DBIL, IBIL, ALT and AST were
detected by the automated biochemistry analyzer as well. RESULTS: The levels of
sCD58 in serum and membrane CD58 molecule in PBMC of patients with hepatitis B
were significantly higher than that in normal controls (P < 0.05). Level of CD58 was
related to the levels of serumal TBIL, DBIL, IBIL, ALT and AST. CONCLUSION: The
level of CD58 molecule (in both serum and PBMC form) of patients with hepatitis B
is related to the degree of liver damage.


Systemic lupus erythematosus following virological response to

peginterferon alfa-2b in a transplanted patient with chronic hepatitis
C recurrence
Lodato F, Tame M R, Colecchia A, Racchini C, Azzaroli F, D'Errico A,
Casanova S, Pinna A, Roda E, Mazzella G
World J Gastroenterol
12(26):4253-5, 2006
Autoimmune manifestations are common both in patients chronically infected by
hepatitis C virus, and in patients transplanted for non-autoimmune diseases. A
correlation between interferon based treatment and autoimmune diseases or the
development of autoantibodies is well established in non-transplanted patients, but
few data are available about transplanted patients. It is unclear whether interferon
may increase the incidence of acute cellular rejection and there are few reports on
the development of atypical autoimmune manifestations during post-liver
transplantation interferon or pegylated interferon treatment. We describe a case of
systemic lupus erythematosus following treatment with pegylated interferon alfa-2b
in a transplanted patient with recurrence of chronic hepatitis C. Our experience
suggest that pegylated interferon may induce autoimmune diseases in the
immunosuppressed host, different from acute cellular rejection and call for a great
attention to possible autoimmune disorders development during interferon based
treatments in liver transplanted patients.