Tuesday Posters: HBV Pathogenesis
May 23rd 8:00 AM-5:00
PM
A. Yasuo ; M.
Moriyama; A. Yasuyuki
Aim.
The purpose of this study is to assess the influence of
occult HBV infection on histopathologic impact and
its clinical outcome of chronic hepatitis C according to prospectively, and is
to determine the nucleotide sequence of occult HBV infection.
Materials and Methods.
The subjects were 468 paients with
HCV RNA-positive (C-viral) chronic hepatitis (CH) or liver cirrhosis (LC) who
were negative for serum hepatitis B surface (HBs)
antigen, and underwent liver biopsy from, l993 to 2003 The serum hepatitis B
virus (HBV) DNA was measured by the nested PCR using the primer set of the X
gene region. HBV DNA-positive patients and -negative patients were compared
with clinical profiles, histological findings and long-term outcome. Then, the
serum HBV DNA level of 182 out of 204 patients sera whose serum HBV DNA was
detected by nested PCR were measured using the real-time detection direct test.
Further, we determined the nucleotide sequence of 2nd PCR products, and
analyzed HBV genotype by phylogenetic tree anlysis.
Results.
The serum HBV DNA was detected in 204 (43.6%) of the 468
patients studied. There were no significant differences between HBV
DNA-positive patients and -negative patients concerning clinical profiles. HBV
DNA-positive patients of the degree of the inflammatory cell infiltration and irregular
regeneration of hepatocytes showed significantly stronger than those of HBV
DNA-negative patients. Serum HBV DNA levels in 169 patients sera (91.8%) showed
less than 1.0 log IU/ml. 15 patients sera whose HBV DNA levels more than 1.0
log IU/ml, and were classified 11 for 1.0 to 1.9 log IU/ml, and others were 2.1
log IU/ml, 3.0 log IU/ml, 3.5 log IU/ml and 5.3 log IU/ml. Cumulative
probability of hepatocellular carcinoma (HCC) incidence of HBV DNA-positive
patients were significantly higher than HBV DNA–negative patients. Furthermore,
HBV DNA-positive was risk factor for occurrence of HCC from C-viral liver
disease by multivariate regression analysis. Alignment of the nucleotide
sequences of HBV genotypes A to F and the 6 NU isolates revealed the nucleotide
substitutions nt 12 in C to T, and nt 14 in G to A. The HBV isolates from these patients were
classified as HBV genotype C by phylogenetic
analysis. The HBV isolates from these patients were clustered in the same group
as isolates from the Japanese mainland.
Conclusion.
The histopathologic impact and
long-term outcome of C-viral CH or LC could be affect by occult HBV infection.
The HBV isolates from these patients were classified as HBV genotype C
T1855. Mutations of hepatitis B virus surface gene on
T cell epitopes in children with chronic infection and hepatocellular
carcinoma.
Y. H. Ni; M. Chang; H. Hsu; H. Chen
Hepatitis B surface antigen (HBsAg)
is an antigen the host immune response attack against in hepatitis B virus
(HBV) infection. This study aims to investigate the mutation(s) of HLA-A2
restricted T cell epitope (TCE) on HBsAg and its relation to chronic HBV infection. Totally
441 carrier children were followed-up for more than 10 years. TCE mutation was
examined at the latest follow-up. They were divided into hepatitis e antigen
(HBeAg) (-) (n=60) and HBeAg (+) groups based on their initial status. HBeAg
(+) group was further divided into HBeAg (+/+) group (n=152) and HBeAg (+/-)
group (n=229) if spontaneous HBeAg seroconversion occurred. Another 25 children
with HBV-related hepatocellular carcinoma (HCC) were also enrolled. The TCE
mutations, liver enzymes, genotypes, HLA, and vaccination histories have been
correlated. HBeAg (+/-), HBeAg (-), and HCC groups had a similar TCE mutation rate,
(26.7%-29.2%), which was higher than that of HBeAg (+/+) group (7.9%). In HBeAg
(+/-) group, 11.9% (8/67) already had such mutations existing before HBeAg
seroconversion. Ten mutations spanning in codons
31-51 of HBsAg were found, while codons
40 and 44 were two hot spots. Those with TCE had an older age of HBeAg
seroconversion than those without (16.1±5.3 vs. 12.7±5.7 years p<0.0001.)
Sixty-six of the 441 subjects received HBV immunization but failed. TCE
mutation rate was higher in the unimmunized patients
than the immunized patients. Genotypes and HLA did not play major roles in the
emergence of TCE mutation. In conclusion, the TCE mutations in children may
develop before HBeAg seroconversion, but it mostly occurred after HBeAg
seroconversion.
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J. Wang; H. Wang; C. Zhu
Objectives
To investigate the relationship between clinical/pathological
features and development of cirrhosis in patients with chronic hepatitis B
(CHB). To determine the time to progress(TTP) and the prognostic risk factors
for developing cirrhosis in patients with HBeAg negative CHB (e-CHB) compared
with HBeAg positive CHB (e+CHB).
Methods
Data collected in consecutive CHB patients confirmed by liver
biopsy (informed consent signed)within 1 week after hospitalized in Zhongshan Hospital, Shanghai from May, 1993 to Jan. 2004
and followed up until June, 2004. Patients received antiviral treatment were
excluded. Scheuer classification has been used for
grading and staging of liver specimen by one pathologist. The difference of
grade and stage, and difference of TTP between e-CHB patients and e+CHB patients were assessed by rank sum test and life
table with Gehan test respectively. Cox regression
model was used to determine the prognostic risk factors associated with developing
compensated and decompensated cirrhosis.
Results
434 patients met the inclusion criteria, 52 (12%) lost follow
up. 382 patients (male 315 and female 67; age 15-71 yr with mean 33.57 yr) were
included in this study. After the follow-up period of an average of 88.64 weeks
(20-260 weeks), 75 cases (19.6%) occured compensated
cirrhosis with an annual occurrence rate of 0.95%. 32 cases (8.4%) progressed
to decompensated cirrhosis with an annual occurrence rate of 0.39%.
120 patients (M 106, F 14 ; age: 34.3yr) were e-CHB, 262 were
e+CHB (M 209, F 53, age 32.93yr). The cases developed
compensated cirrhosis in e-CHB group compared to e+CHB
was 22 (22.5%) vs 48 (18.3%) respectively. The cases
progressed to decompensated cirrhosis were 14 (11.61%) in e-CHB group and
18(6.87%) in e+CHB group respectively. There is no
statistically significant difference between these two groups for grade and
stage by rank sum test as well as cirrhosis development showed in life table by
Gehan test. The result of multivariable analysis by
Cox regression model also indicates that histological staging at
hospitalization is independent prognostic factor for developing compensated and
decompensated cirrhosis in CHB (P<0.01, HR: 2.14 and 1.7). Histological
grade, serum albumin, globulin, AST, ALT, AFP, PT tested at hospitalization,
age, sex, history of HBV, e-CHB were not associated with development of
cirrhosis in this model.
Conclusion
The severity of the fibrosis stage at presentation is closely
related with compensated and decompensated cirrhosis development in CHB rather
than severity of inflammatory grade and e-CHB.
T1857. HBV vaccination with evidence of protective
antibodies titer induces CD4+/CD25+ T cells
A. Perrella;
L. Racioppi; L. Atripaldi;
S. Pisanti; C. Sbreglia; P.
Bellopede; O. Perrella
Background/Aim:
The immunoregulatory mechanisms
occurring after the HBV vaccine administration are not well known. Aim of the
present study was to enumerate CD3+/CD4+/CD25+ T cells and to test their
function after HBV vaccination at the seroconversion of HBsAg
comparing them to patients with chronic hepatitis B.
Methods:
We enrolled 10 subjects undergoing three doses of HBsAg vaccine (Engerix-B
DNA-recombinant)injected intramuscularly in the deltoid region at time
0(T0),one(T1) and sixth month(T6), following the manufacturer’s instructions
(Group A) and 10 healthy donors (Group B) (table). CD3+/CD4+/CD25+ lymphocytes
were enumerated in PBMC using flow cytometry (Beckman
Coulter). T lymphocytes activity was assessed using immunomagnetical
sorting with magnetic bids (purified Tregs > 95%)
(Dynal Treg kit) as
capacity to inhibit the proliferation of CD4+/CD25- T cells stimulated with
anti-CD3/CD28 (1:2 and 1:20 ratio).
Results:
Subjects were administered vaccine after one month achieved a
protective titer of Anti-HBs
(> 100IU/mL) and had a contemporary statistically significant increase of
CD4+/CD25+ frequency and function when compared to patients suffering chronic
hepatitis B and their self before the administration of first dose of vaccine
(U Mann-Whitney Test z= 2.23; p < .05 and z= 2.22; p< .05)(table).
Conclusion:
An increased CD3+/CD4+/CD25+ T cells frequency and function
seems to occur after one month of vaccine administration when a protective titer is achieved, suggesting a role of CD4+/CD25+ T
regulatory cells in the seroconversion and proposing them as a possible tool to
monitor antiviral treatment response.
T1858. Novel Subgenotypes of Hepatitis B Virus Among Chronic Liver Disease
Patients in the Philippines
T. Sakamoto; Y. Tanaka; E. Orito; J. Co; J. Clavio; F. Sugauchi; K. Ito; A. Ozasa; A. Quino; R. Ueda; M. Mizokami; J. Sollano
Several hepatitis B virus (HBV) subtypes (subgenotypes),
HBV/Aa (Asia/Africa), Ae
(Europe), Ba (Asia) and Bj
(Japan) were reported with respect to the clinical differences between the
patients infected with these subtypes. There have been no investigative
population studies on HBV genotypes in the Philippines. We investigated the HBV
genotype distribution among patients with chronic liver diseases (CLD) in the
Philippines. One hundred sera were obtained from CLD patients, consisting of 32
chronic hepatitis (CH), 37 cirrhosis and 31 hepatocellular carcinoma (HCC).
Nine complete genomes and 100 core promoter/precore
genes of HBV were sequenced directly Phylogenetic
analyses showed that 51 HBV/Aa, 22 HBV/Ba and 27 HBV/C strains were found in this study.
Interestingly, most HBV/C strains in the Philippines had the specific cluster
distinct from previous HBV/C strains (C1-4), indicating a novel subtype, HBV/Cp
(p standing for Philippines; C5). Moreover, most HBV/B strains had the specific
cluster as the fifth HBV/B subgroup (B5) with viral characteristics of HBV/Ba. In the 3 genotypes, HBV/Ba or
HBV/C were significantly more prevalent in cirrhosis and HCC patients than HBV/Aa (p<0.02). The prevalence of core promoter (CP)
mutation (T1762/A1764) was more frequent in HCC patients with HBV/Ba and HBV/C. Multivariate analysis indicated that age (OD
3.43; 95%CI 1.04–11.36; p=0.044) and CP mutation (OD 14.08; 95%CI 3.62–4.74;
p<0.001) were significant factors for HCC development. In conclusion, novel
subtypes Cp and B5 are prevalent in the Philippines, as well as HBV/Aa.
T1859. Association between Hepatitis B Virus Infection
and HLA-DRB1 Genotyping in Shaan’xi Han patients in Northwestern
China
J. Liu; G. Yang; H. Xie;
S. Han; R. Du; D. Fan
AIM:
HBV infection is a major public health problem worldwide,
especially in China, where 7±20% of the large population is chronically
infected with estimated millions. The mechanism of susceptibility to chronic
persistent hepatitis B virus (HBV) infection is not well clarified, while the
outcome of HBV infection mainly depends on the host immune response. HLA class
II molecule is an integral component of the immune response on which majority of
host genetic studies have concentrated. Many different HLA class II alleles
have been demonstrated to play roles in the HBV infection. In this study, the
association of HBV infection and HLA-DRB1 allele in Han individuals in Western
China population were studied for the first time.
METHODS:
216 Shaan’xi Han individuals were
categorized into 2 different groups: the HBV patients group (n=108), and health
control group (n=108). HLA Class II types were determined by PEL-FREEZ® SSP UniTray® which is a PCR-based method designed to provide
low to high resolution of the various HLA types.
RESULTS:
DRB1*04, DRB1*09, DRB1*12, DRB1*15 were the most common
genotypes in all groups with the frequencies of 16.2%,12.5%,11.6% and 13.4%
respectively. The allele frequencies of HLA-DRB1*03 (10.6% of HBV patients
versus 3.7% of health controls, odds ratio=3.10; Pc=0.008; P<0.05) and
HLA-DRB1*07 (17.6% of HBV patients versus 9.3% of health controls, Pc=0.016;
Odd ratio=2.09; P<0.05) were markedly higher in HBV group. The allele
frequency of HLA-DRB1*15(6.9% of HBV patients versus 13.4%of health controls
Pc=0.039; Odds ratio=0.48; P<0.05) was obviously lower than that in
controls. It is indicated that HLA-DRB1*03 and HLA-DRB1*07 are related with
susceptibility to chronic hepatitis B, and DRB1*15 is closely related with
resistance to chronic hepatitis B in Northwestern
China. Similar results was got on DRB1*03 and 15 alleles between the patients
(n=108) and 46 HBV seronegative spouses of HBV
patients, which was a high-risk group for HBV infection.
CONCLUSION:
All these suggest that host HLA classII
gene is an important factor determining the outcome of HBV infection.
T1860. Hepatitis B genotype Bj in a
cohort of Alaska Native People with chronic infection.
J. Simonetti;
M. Snowball; S. Negus; C. Homan; J. Williams; Y. Tanaka; M. Mizokami;
B. McMahon
Introduction
Hepatitis B (HBV) has been classified into eight genotypes
(A-H) with distinct geographical distributions and differing influences on the
outcome of chronic infection. Recently, recombination between genotypes has
been reported and is emerging as an important factor in the outcome of HBV
infection. Further investigation has revealed that the Taiwanese genotype B (or
Ba) harbors a recombination
with genotype C over the precore region plus the core
gene while the Japanese B (or Bj) does not harbor a recombination and has a less severe disease
course. Bj is found exclusively in Japan while Ba is ubiquitous in the other countries in Asia. In
addition, HBeAg positivity and the basal core
promoter mutation (T1762/A1764) are found to be more frequent in Ba than Bj. A long-term study of
Alaska Native People with chronic HBV infection has identified 5 genotypes with
genotype B constituting 4% (46/1188) of the total number. In this preliminary
study we further identified the subtype of genotype B in this cohort.
Methods
Genotypes Ba and Bj were identified through semi-nested PCR and direct
sequencing of a 700-bp region of the precore region
plus the core gene. Serum with sufficient HBV DNA was available for 40 of the
46 original genotype B patients. HBV DNA levels were determined using a
Real-time Quantitative PCR assay.
Results
Of the 40 genotype B patients tested 100% were classified as Bj. The median age of the genotype B patients is 55 and to
date hepatocellular carcinoma (HCC) has been diagnosed in only one individual
(a 76-year old male) from this group and none have been diagnosed with chronic
hepatitis. Furthermore, the genotype B patients in this cohort maintain consistently
low (< 20,000 IU/ml) HBV DNA levels and rarely have elevated liver function
tests.
Conclusion
Genotype Bj is the exclusive
genotype B subtype found in a cohort of Alaska Native People with chronic HBV.
In addition, the disease course of patients infected with genotype Bj mirrors the less severe course observed in Japan.
J. M. Mellen;
N. L. Kyulo; V. Xia; J. Hoefs; D. Imagawa; K. Hu
BACKGROUND:
The clinical course of HBV infection varies with ethnicity.
Although numerous studies have focused on the natural history of this disease
in the Asian population, little is known about the clinical presentation of
chronic HBV infection in Asian Americans.
AIMS:
To define the natural history of HBV infection in Asian
Americans.
METHODS:
This was a retrospective study by chart review of > 300
consecutive patients presented to the University of California Irvine Medical
Center between Jan. 1990 and Nov. 2005 with a diagnosis of HBV infection. The
inclusion criteria were documented Asian ethnicity, positive HBsAg or HBV DNA for 6 months, no prior HBV treatment at
presentation, and no HCV or HIV co-infection. After excluding 140 cases who did
not meet the inclusion criteria, 172 patients were included in the study.
RESULTS:
This cohort of patients had a mean age of 41.9 + 15.0,
contained 66.9% male patients, 97.8% were born outside of the U.S., and 13.4%
had histological diagnosis of hepatocellular carcinoma (HCC). Family history of
hepatitis B was reported in 47.7% of the patients, but history of injection
drug use (2.5%), blood transfusion (14.4%), and current alcohol use (12.1%)
were less common. Of the 57 patients with liver biopsy, 42.1% had stage 3-4
fibrosis. In 14 patients with HCC, stage 3-4 fibrosis was present in 78.6% of
non-tumorous liver tissue, and this was significantly
higher than in patients without HCC (30.9%, p=0.004). Stage 3-4 fibrosis was
not significantly associated with the baseline ALT value (p=0.76), HBV DNA load
(p=0.78), steatosis (p=0.21). The mean body mass index (BMI) was 23.8 + 3.4
kg/m2, and steatosis was reported in 13.0% of the patients. The incidence of
HBeAg negativity was as high as 79.5% in this cohort of patients.
HBeAg-negative HBV infection was associated with elder age (p=0.05), no known
family history of hepatitis B (p=0.009), non-elevated ALT (p=0.001), and a
lower HBV DNA load (p=0.0003). However, HBeAg status was not associated with steatosis
(p=0.56), stage 3-4 fibrosis (p=0.45), or a diagnosis of HCC (p=0.46). A
multivariate analysis indicated that after adjusting for age and family history
of hepatitis B, a low HBV DNA load and a non-elevated ALT were independently
associated with HBeAg-negative HBV infection.
CONCLUSIONS:
This single center study indicated that HBeAg-negative HBV
infection is a common presentation in Asian Americans, and that it is
independently associated with a low HBV DNA load and a non-elevated ALT. Asian
American patients with chronic HBV infection carry a high incidence of advanced
fibrosis and a low incidence of steatosis.
T1862. Occult hepatitis B virus infection with and
without anti-HBs antibodies - implications in the development of
hepatocellular carcinoma C.
Tan; J. Chang; G. Lim; Z. Yi; L. Ooi; P. Chow; A. Chung; W. Chow
Background:
Occult hepatitis B virus (OHBV) infection is defined as the
detection of HBV DNA in serum or liver tissue of HBsAg
negative patients. Patients with OHBV infection who are anti-HBc positive can be further differentiated into those with
and without anti-HBs antibodies (anti-HBsAb). Those with anti-HBsAb may
be regarded as having evidence for immunoclearance of
HBsAg and hence have an advantage over those without
anti-HBsAb who are often regarded as low grade viraemic carriers. Hepatocellular carcinoma (HCC)
uncommonly occurs in cases of OHBV infection and HBV DNA may be detected in the
liver and tumor tissues. However, it is not known
whether the extent of tissue HBV integration differs between cases with and
without anti-HBsAb.
Aim:
To study whether the extent of HBV integration in tumor and non-tumor tissues is
influenced by presence of anti-HBsAb in HCC patients
with occult HBV infection.
Methods:
Study subjects were consecutive HBsAg-negative,
anti-HBc positive HCC patients who underwent surgical
resection at our centre. Our IRB approved the study and all patients gave
informed consent. Patients were grouped based on anti-HBsAb
status – Group 1 (anti-HBsAb positive) and Group 2
(anti-HBsAb negative). Nested PCR techniques were
used to determine presence of HBV “S” and “X” genomes in tumor
and non-tumor tissues and serum. Ten HBsAg positive and 5 seronegative
(HBsAg/anti-HBc/anti-HBs/serum HBV DNA) patients with
resected HCC served as positive and negative controls
respectively.
Results:
There were 6 patients in Group 1 and 5 in Group 2. Mean anti-HBsAb level in Group 1 was 134iu/ml (range 17-564). “S”
genome was detected in 5/6(83%) tumor tissue (TT) and
6/6(100%) non-tumor tissue (NT) in Group 1 and
4/5(80%) TT and 3/5(60%) NT in Group 2. “X” genome was detected in 3/6(50%) TT
and 6/6(100%) NT in Group 1 and 5/5(100%) TT and 3/5(60%) NT in Group 2.
Concordance rates between NT and TT in detection of “S” genome were 83% and 80%
for Groups 1 and 2 respectively; for “X” genome, they were 50% and 60%
respectively. There were no significant differences (Fisher’s test) between the
2 groups in all the above results. As for the sera, nested PCR did not detect
“X’ genome at all in both groups but “S” genome was found in 1 case in Group 1
that also had integration of “S” and “X” genomes in both TT and NT.
Conclusions:
Despite the presence of anti-HBs
antibody, there is a similar extent of genomic integration of HBV into host liver
tissue. This suggests the pathogenesis of HCC is similar in OHBV infection with
or without anti-HBsAb. Therefore, regardless of anti-HBsAb status, strategies to prevent the development of HCC
should be similar.