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During a median period of 22.8 years after HBeAg seroclearance, 40 66% patients became inactive carriers, whereas the remaining 21 34% showed alanine aminotransferase elevation: one 1% h

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doi:10.1136/gut.2007.128496 2008;57;84-90; originally published online 22 Aug 2007;

Gut

G Fattovich, N Olivari, M Pasino, M D’Onofrio, E Martone and F Donato

Caucasian patients: mortality after 25 years Long-term outcome of chronic hepatitis B in

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Long-term outcome of chronic hepatitis B in Caucasian patients: mortality after 25 years

G Fattovich,1 N Olivari,1 M Pasino,1 M D’Onofrio,2 E Martone,2 F Donato3

1 Department of

Gastroenterology, University of

Verona, Verona, Italy;

2 Department of Radiology,

University of Verona, Verona,

Italy; 3 Institute of Hygiene,

Epidemiology and Public Health,

University of Brescia, Brescia,

Italy

Correspondence to:

Dr G Fattovich, Unita ` Operativa

di Gastroenterologia,

Dipartimento di Scienze

Chirurgiche e

Gastroenterologiche, Universita `

di Verona, Piazzale L.A Scuro n.

10, 37134 Verona, Italy;

giovanna.fattovich@univr.it

Revised 5 August 2007

Accepted 15 August 2007

Published Online First

22 August 2007

ABSTRACT Objective: To assess risk factors for liver-related death,

we re-evaluated, after a median follow-up of 25 years, a cohort of 70 Caucasian patients with hepatitis B e antigen (HBeAg) positive chronic hepatitis (CH) at presentation

Methods: Follow-up studies included clinical and ultra-sound examinations, biochemical and virological tests, and cause of death

Results: Sixty-one (87%) patients underwent sponta-neous HBeAg seroconversion During a median period of 22.8 years after HBeAg seroclearance, 40 (66%) patients became inactive carriers, whereas the remaining 21 (34%) showed alanine aminotransferase elevation: one (1%) had HBeAg reversion, nine (15%) detectable serum HBV DNA but were negative for HBeAg, eight (13%) concurrent virus(es) infection and three (5%) concurrent non-alcoholic fatty liver disease Liver-related death occurred in 11 (15.7%) patients, caused by hepatocellular carcinoma in five and liver failure in six The 25-year survival probability was 40% in patients persistently HBeAg positive, 50% in patients with HBeAg negative CH

or HBeAg reversion and 95% in inactive carriers Older age, male sex, cirrhosis at entry and absence of sustained remission predicted liver-related death independently The adjusted hazard ratios (95% CI) for liver related death were 33 (3.01–363) for persistently HBeAg positive patients and 38.73 (4.65–322) for those with HBeAg negative CH or HBeAg reversion relative to inactive carriers

Conclusion: Most patients with HBeAg seroconversion became inactive carriers with very good prognosis The risk of liver-related mortality in Caucasian adults with CH

is strongly related with sustained disease activity and ongoing high level of HBV replication independently of HBeAg status

Chronic hepatitis B virus (HBV) infection is an established cause of liver-related morbidity and mortality and represents a major global public health problem.1 2 Studies conducted in Asian regions of high HBV endemicity have estimated that up to 40%

of individuals with chronic HBV infection will progress to liver cirrhosis, end-stage liver disease or hepatocellular carcinoma (HCC) during their life-time and that HBV-related liver disease contribute to approximately 20 deaths per 100 000 population each year.3 4

The long-term liver-related morbidity and mortality of chronic hepatitis B in untreated Caucasian patients and predictors of survival deserve further evaluation In fact, the prognostic value of sustained high levels of HBV replication on survival

in untreated Caucasian patients is limited to cohorts

of patients with cirrhosis.5–7

We conducted this longitudinal study to eluci-date the long-term survival according to virological

pattern in a cohort of north Italian patients with chronic hepatitis B who had high levels of HBV replication (HBeAg positive) at diagnosis, during a median period of 25 years

PATIENTS AND METHODS Patients

A longitudinal study began in Padova, northern Italy, enrolling between 1972 and 1984 seventy consecutive hepatitis B surface antigen (HBsAg) and HBeAg positive patients, 49 male and 21 female with a median age of 29 years and with biopsy proven chronic hepatitis (8 (11.4%) with cirrhosis) at presentation The clinical, virological and histological details of the study population at entry and the natural history of the disease during

a mean follow-up period of 5.0 years (SD 2.3 years) were described previously.8 The follow-up of this cohort of patients has been performed at the Department of Gastroenterology at Verona University Hospital in northern Italy and updated through June 2006

Follow-up data were obtained during regular clinical controls, on average every 6 months for patients who remained HBeAg positive or sero-converted from HBeAg to anti-HBe, but main-tained or developed abnormal ALT levels and on everage every 12 months for inactive HBsAg carriers By 1986 it became clear that HBeAg seroclearance with ALT normalisation is usually associated with histology improvement,8 thus in the subsequent years follow-up liver biopsies were performed only in patients with active hepatitis Patients who did not undergo the regular controls were traced through the municipality registries and were phoned and asked to attend again the clinic for evaluation People who refused to attend the clinic again were interviewed by phone and asked

to provide laboratory and liver ultrasound exam-inations with the consent of their family doctors Data on patients’ liver-related or other diseases that had been collected through the phone inter-views were cross-checked with the archives of their family doctors The causes of death for those deceased were ascertained by retrieving the death certificates We were able to trace all patients except two (97.1%) from the initial cohort The current study was approved by the Verona Hospital Ethics Committee

Definition of clinical events The time of observation was calculated from the date of entry into the study until death, liver transplantation or the last observation The primary outcome was liver related mortality The occurrence of cirrhosis during follow-up was

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defined by histology or ultrasonography features suggestive of

cirrhosis based on a quantitative scoring system derived from

the appearance of the liver margins, parenchymal echotexture,

portal vein calibre and spleen diameter,9 10supplemented with

the presence of oesophageal varices or ascites HCC diagnosis

was based on the following criteria: a histopathological

examination; a positive lesion detected by at least two different

imaging techniques (abdominal ultrasonography, computerised

tomography, or angiogram); a positive lesions detected by one

imaging technique and a-fetoprotein levels exceeding 400 mg/l

Biochemical remission was defined as the normalisation of

serum ALT levels on at least two consecutive determinations

obtained at least 3 weeks apart and persisting at last

observa-tion After HBeAg seroclearance with anti-HBe seroconversion,

patients were defined as inactive HBsAg carries when ALT

values were persistently normal and HBV DNA was persistently

negative according to non-polymerase chain reaction (PCR)

assays (detection limit, 1.46105 copies/ml) Active hepatitis,

after HBeAg seroclearance, was defined as ALT elevation over

twice the upper normal value on at least two consecutive

determinations obtained at least 3 weeks apart HBeAg negative

hepatitis was diagnosed when abnormal ALT and HBV DNA

seropositivity by non-PCR assays persisted continuously or

recurred intermittently after HBeAg seroclearance

Laboratory procedures

HBsAg, antibody to HBsAg (anti-HBs), HBeAg, antibody to

HBeAg (anti-HBe), hepatitis delta virus (HDV) and hepatitis C

virus (HCV) antibodies, hepatitis A virus IgM antibodies were

detected by conventional serological assays Serum HBV DNA

was first tested by homemade spot hybridisation with a

detection limit of 0.1 pg/ml,8then the Digene Hybrid Capture

System assay (Digene, Beltsville, MD) was used, with a

detection limit of 0.5 pg/ml (1.46105copies/ml) More recently

HBV DNA was tested with a commercial PCR assay (Cobas

Amplicor HBV Monitor, Roche, Basel, Switzerland) with a

detection limit of 200 copies/ml

Serum HDV RNA was detected by an ‘‘in-house’’

reverse-transcriptase PCR (RT-PCR) in the virology laboratory of the

University Hospital of Verona Briefly, HDV RNA was

extracted from 200 ml of serum, reverse transcribed and

amplified by one-step RT-PCR in a final volume of 50 ml using

a set of primers derived from the carboxyterminal portion of the

genomic region coding for the HDV antigen (primers 59 R 39,

position 886–909 and 1331–1310, respectively) The

amplifica-tion products were visualised on a 2% agarose gel stained with

ethidium bromide For each test sample a negative and a

positive control were included in parallel throughout the entire

procedure Serum HCV RNA was first measured by an

‘‘in-house’’ nested RT-PCR with a detection limit of 100 copies/

ml,11then was assessed using a commercial PCR assay (Cobas

Amplicor HCV Monitor, Roche, Basel, Switzerland)

Histological diagnosis was defined according to international

criteria.12 13

Statistical analysis

The Mann–Whitney and x2 and exact tests were used for

comparison of continuous and categorical variables between

two independent groups p values lower than 0.05 in two-tailed

tests were considered as significant

The cumulative probabilities of survival were analysed by the

Kaplan–Meier method14 and comparison between groups was

performed by using the log-rank test.15Patients who died from

causes that were not related to liver disease or were lost to follow-up or responded to antiviral therapy were censored at the time of death, drop out or response to antiviral therapy, respectively Multivariate analysis was also performed by fitting the Cox proportional hazard models in order to evaluate the role

of each prognostic factor when controlling for the others The assumption of proportionality of hazards functions was assessed using graphical methods as suggested.16

Statistical analyses were performed using the STATA soft-ware (Stata Statistical Softsoft-ware release 8.0, 2003; Stata Corporation, College Station, Texas)

RESULTS Serological outcome HBeAg seroconversion Our cohort of 70 HBeAg positive patients was followed for a median period of 25.3 years (range, 1.3–33.1) for a total of 1564 person-years During this observation period nine (12.8%) patients remained persistently HBeAg positive and 61 (87.2%) underwent spontaneous HBeAg seroclearance with anti-HBe seroconversion (fig 1) The cumulative probabilities of HBeAg seroconversion were 64%, 93% and 97% at 5, 10 and 25 years of follow-up, respectively Most patients underwent HBeAg seroconversion during the first 10 years of the study (58 of 61, 95.1%), with an incidence rate of 18.7 per 100 person-years At enrolment, patients who remained persistently HBeAg positive were older (median age, 51 vs 27 years) and had a higher prevalence of cirrhosis (44% vs 6.6%) as compared with those who underwent spontaneous HBeAg seroclearance (table 1) At the time of HBeAg seroconversion, their median age was 30 years (range, 16–65) and 11 (18%) had histological evidence of cirrhosis The 61 patients with HBeAg seroclearance were followed for

a median period of 22.8 years (range, 0.5–28.4) after HBeAg seroconversion Forty (66%) out of the 61 patients had sustained ALT normalisation and undetectable HBV DNA by non-PCR tests and were considered inactive carriers; more recently HBV DNA was tested by PCR assay in 15 (37.5%) out

of the 40 inactive carriers and was detectable in all cases ranging from 700 to 9800 copies/ml The remaining 21 (34%) out of the

61 patients experienced active hepatitis; among these 21 patients, ALT elevation was associated with HBeAg reversion

in one (1%), detectable serum HBV DNA but HBeAg negative (HBeAg negative hepatitis) in nine (15%), concurrent HCV and/

Figure 1 Study flow diagram NAFLD, non-alcoholic fatty liver disease

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or HDV infection in eight (13%) and concurrent non-alcoholic

fatty liver disease (NAFLD) in three (5%) cases (fig 1) Of the

three patients with NAFLD, two had histological features of

non-alcoholic steatohepatitis (NASH) and one was obese (BMI

38 kg/m2) with fatty liver diagnosed by ultrasound All three

cases with NAFLD were seronegative for both HBeAg and HBV

DNA by non-PCR tests; at the last visit HBV DNA was

detectable by PCR ranging from 300 to 8750 copies/ml

The clinical and biochemical features of the 61 patients with

spontaneous HBeAg seroconversion according to clinical and

serologic categories during follow-up (40 with sustained

remission and 21 with active hepatitis) are given in table 2

The patients who developed HBeAg negative hepatitis or

HBeAg reversion had a higher prevalence of cirrhosis at the

time of HBeAg seroconversion compared to patients with

sustained remission (50% vs 17.5%, respectively; p = 0.04)

(table 2)

HBsAg loss

Eighteen (45%) of the 40 inactive carriers and one (12.5%) of the

eight patients with viral concurrent infections lost their HBsAg

and developed the related antibody (table 2) The incidence of

HBsAg loss was 2.1 per 100 person-years in inactive carriers

Among the 18 inactive carriers with HBsAg loss the median

interval between HBeAg seroconversion and HBsAg clearance

was 13.8 years (range, 1.1–26.9)

Clinical outcome

Patient survival

A total of 50 patients were alive at the end of the follow-up:

among them, 30 (60%) were checked into the hospital, 18 (36%)

were interviewed by phone and two were lost at the follow-up

Liver-related death occurred in 11 (15.7%) patients, caused by

HCC in five and liver failure in six, and one patient underwent OLT for end-stage liver disease (table 1)

The cumulative probability of survival among all patients was 90% and 86% at 10 and 25 years of follow-up, respectively The cumulative probability of survival at 25 years was significantly lower in the nine patients who remained HBeAg positive (40%) and in patients developing HBeAg negative hepatitis or HBeAg reversion (50%) as compared to inactive carriers (95%) (p,0.0001) (fig 2) None of the patients with concurrent HCV and/or HDV infection or patients with concurrent NAFLD died during the 25 years of observation The cumulative probability of survival at 25 years was significantly lower in eight patients with cirrhosis at entry (22.5%) and in 17 patients who developed cirrhosis during follow-up (59%) as compared to 45 patients without cirrhosis (100%) (p,0.0001)

Cox proportional hazard regression model showed that older age at diagnosis, male sex, the presence of cirrhosis at enrolment and the absence of sustained remission during follow-up were significantly associated with an increased risk for liver related death or OLT (table 3) In a subgroup analysis excluding patients with concurrent HCV and/or HDV infection or NAFLD, the risk of liver related mortality or OLT was increased 33-fold in patients with HBeAg persistence and 38-fold in those with HBeAg negative hepatitis or HBeAg reversion as compared

to patients with sustained remission (inactive carriers) (table 3)

Antiviral therapy Ten patients with evidence of high levels of HBV replication (two HBeAg positive and eight with HBeAg negative hepatitis) were still alive in the late 1980s when interferon (IFN) therapy became available and three of them (one HBeAg positive and two HBeAg negative) were treated with IFN An additional four out of the eight patients with concurrent viral concurrent

Table 1 Demographic and clinical features of different subsets of the study population at enrolment and during follow-up

All patients HBeAg persistence HBeAg seroclearance

Features at entry Age (years) (median, range) 29 (15–64) 51 (26–64) 27 (15–62)

ALT 6 normal (median, range) 2.9 (2–16) 2.7 (2–4.2) 3.0 (2–16) Previous acute hepatitis 30 (42.9) 4 (44.4) 26 (42.62) Presumed source of infection

Medical occupation 13 (18.6) 0 13 (21.3) Household contact 17 (24.3) 0 17 (27.9)

Presence of cirrhosis 8 (11.4) 4 (44.4) 4 (6.6) Features during follow-up

Cirrhosis occurrence 17 (24.3) 4 (44.4) 13 (21.3)*

Total number of deaths 19 (27.1) 7 (77.8) 12 (19.7) Liver-related deaths 11 (15.7) 5 (55.6) 6 (9.8)

Non-liver related deaths 8 (11.4) 2 (22.2) 6 (9.8)

Data expressed as number (%).

ALT, alanine aminotransferase; OLT, orthotopic liver transplantation.

*Seven patients developed cirrhosis before HBeAg seroclearance and six patients after HBeAg seroclearance.

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infections received antiviral therapy Among the seven treated

patients, two (one HBeAg positive and one HBV/HCV

co-infected patient) responded to antiviral therapy and were

censored from follow-up The details of IFN schedule and

reasons for not treating the remaining patients are provided in

the following sections

Clinical outcome of HBeAg positive patients

Among the nine patients who remained HBeAg positive during

a median follow-up period of 6.8 years (range, 1.3–25), eight had

histologically documented cirrhosis (four diagnosed at entry and

four during follow-up) One patient (with cirrhosis diagnosed during follow-up) responded to lymphoblastoid IFN at the dose

of 8 million units thrice weekly for 6 months in the year 1989 The remaining seven patients with cirrhosis died: four for decompensated cirrhosis, one for HCC and two for liver unrelated causes Among these seven patients, six died in the early 1980s before antiviral therapy became available and one did not receive IFN therapy due to ischaemic heart disease which was the cause of unrelated death in 1999 Only one (1.4%) of the 70 initially HBeAg positive patients is still HBeAg positive and alive without clinical and/or serological evidence of

Table 2 Clinical and biochemical features in 61 patients with spontaneous HBeAg seroconversion according to clinical and serological categories during follow-up (40 with sustained remission and 21 with active hepatitis)

Sustained remission

HBeAg-/HBV-DNA+

or HBeAg reversion

HDV and/or HCV concurrent infection NAFLD

Features at entry

Age (years) (median, range)* 29.5 (15–52) 24 (15–62) 23.5 (15–39) 15 (13–21)

ALT 6 normal (median, range)* 3.4 (2–10.9) 2.5 (2–9.2) 3.1 (2–16) 2.1 (2–2.4) Features during follow-up

Follow-up after HBeAg seroconversion (years) (median, range){ 23.0 (3.2–28.2) 18.2 (2.8–25.9) 19.4 (0.5–28.4) 20.8 (20.8–24.3) Age at HBeAg seroconversion (year) (median, range)* 33 (17–59) 28 (17–65) 30.5 (19–31) 22 (16–25)

Clinical and laboratory assessment 15 (37.5) 4 (40) 6 (75) 3 (100)

Data are expressed as number (%).

HDV, hepatitis delta virus; HCV, hepatitis C virus; NAFLD, non-alcoholic fatty liver disease; ALT, alanine aminotransferase; HCC, hepatocellular carcinoma; OLT, orthotopic liver transplantation.

Comparison between 40 patients with sustained remission and 10 patients with HBeAg negative hepatitis (HBeAg2/HBV DNA+) or HBeAg reversion (HBeAg+/HBV DNA+): *Not significant; {p = 0.05; {p = 0.04; 1p = 0.002.

Figure 2 Cumulative probability of

survival in the three subsets of patients:

inactive carriers, patients with HBeAg

negative hepatitis or HBeAg reversion and

patients persistently HBeAg positive

(p,0.0001, log-rank test)

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cirrhosis at the last observation in June 2006; the patient had

ALT fluctuating from normal to 1.5 times the upper normal

value during follow-up and remained untreated

Clinical outcome of inactive carriers

The clinical outcome of the 40 inactive carriers during a median

follow-up period of 23 years (range, 3.2–28.2) after HBeAg

seroconversion is shown in fig 3 Seven patients had

histolo-gically documented cirrhosis, three at enrolment and four

progressed to cirrhosis during the HBeAg positive phase before

HBeAg seroconversion Of these seven inactive carriers with

cirrhosis, two died from HCC 7.7 and 9.4 years after HBeAg

seroconversion, respectively, three died of liver-unrelated causes,

one is alive and one was lost to follow-up Of the 33 inactive

carriers without cirrhosis, three died of liver unrelated causes

and the remaining 30 are alive

Clinical outcome of patients with active hepatitis after HBeAg

seroclearance

The clinical outcome of the 21 patients with HBeAg

seroclear-ance and active hepatitis is illustrated in fig 4

One patient with cirrhosis at enrolment showed HBeAg

reversion associated with persistent ALT elevation 15 months

after HBeAg seroconversion and died of liver failure 3.9 years

after sustained HBeAg reversion

Among the nine patients who progressed to HBeAg negative

hepatitis two had histologically documented cirrhosis at the

time of HBeAg seroconversion and two developed clinical

cirrhosis 3 and 13 years after HBeAg seroconversion,

respec-tively Among the four patients with cirrhosis, one died of liver

failure in the year 1981, one did not respond to lymphoblastoid

IFN (8 million units thrice weekly for 6 months) in the year

1987 and underwent OLT 11 years later and the remaining two

patients remained untreated because of advanced cirrhosis with

low platelet levels at the time IFN therapy became available and

died of HCC All five patients with HBeAg negative hepatitis

without cirrhosis are alive All five patients underwent

follow-up liver biopsies after development of HBeAg negative hepatitis

showing mild to moderate chronic hepatitis without cirrhosis

and none showed liver surface nodularity or other

ultrasono-graphic markers of advanced liver disease at follow-up

ultrasound examination One patient failed to respond to

lymphoblastoid IFN (8 million units thrice weekly for

6 months) and the other four remained untreated because of mild histology and no clinical evidence of progressive disease

Of the eight patients with concurrent viral infections, five developed cirrhosis (histologically confirmed in four cases) during follow-up (one during the HBeAg positive phase and four after HBeAg seroconversion) and three had no clinical, liver ultrasonographic and/or biochemical evidence of cirrhosis at the last observation Of the five patients with cirrhosis, one HBV/ HCV infected patient responded to standard combination treatment with pegylated IFN-a and ribavirin, two anti-HDV/ anti-HCV positive (HCV RNA negative) patients failed to respond to recombinant IFNa2a (10 million units thrice weekly for 12 months), one anti-HDV/anti-HCV positive (HDV RNA, HCV RNA and HBV DNA negative by PCR at last observation) patient admitted alcohol abuse and was advised to abstain from alcohol intake and the remaining anti-HDV positive (HDV RNA negative at last observation) patient showed intermittently slightly elevated ALT and remained untreated Of the three patients without cirrhosis, one anti-HDV positive patient was lost to follow-up, one HBV/HCV infected patient lost HBsAg and refused antiviral therapy for HCV and the remaining HBV/ HCV infected patient failed to respond to standard combination therapy with recombinant IFN-a and ribavirin Excluding one anti-HDV positive patient lost to follow-up and one anti-HCV positive patient who responded to antiviral therapy and censored from follow-up, the remaining six patients are alive and without evidence of hepatic complications in those with cirrhosis

None of the patients with active hepatitis after HBeAg seroclearance received nucleoside or nucleotide therapy

DISCUSSION The major aim of this longitudinal study was to assess the prognostic significance of sustained high HBV replication levels

on the risk of liver-related death in a cohort of 70 Caucasian patients with HBsAg and HBeAg positive chronic hepatitis at diagnosis who were followed for a median period of 25 years with a very low drop-out rate The number of patients is relatively small, but the study population consists of a well-defined cohort of consecutive patients previously described,8

thus minimising the risk of selection bias Data on liver-related

Table 3 Hazard ratios (HR) and 95% confidence intervals (CI) for liver related death by Cox proportional hazard regression models (all variables included in the model)

No liver-related deaths/OLT HR (95% CI) p Age at entry (years) 70 12 1.10 (1.06 to 1.16) ,0.001 Sex

Duration of HBeAg positive phase (years) 70 12 0.84 (0.63 to 1.10) n.s.

Cirrhosis at entry

HBeAg2/HBV DNA+ or HBeAg reversion 10 5 38.73 (4.65 to 322.9) 0.001 HBeAg persistence 9 5 33.06 (3.01 to 363.0) 0.004 n.s., not significant.

*The model included the 40 patients with sustained remission (reference category), the 10 patients with HBeAg2/HBV DNA+

chronic hepatitis or HBeAg reversion and the nine patients with HBeAg persistence, and adjusted for age, sex and cirrhosis at entry.

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mortality were accurately obtained in the great majority (97%)

of patients After 25 years, survival was significantly worse in

patients who remained HBeAg positive, developed HBeAg

negative hepatitis or HBeAg reversion, as compared with

inactive carriers (40%, 50% and 95%, respectively) It is of note

that the adjusted hazard ratios for liver-related death were

33-fold higher in patients who remained HBeAg positive and

38-fold higher in HBeAg negative hepatitis who had HBV DNA

positivity or HBeAg reversion with respect to inactive carriers

To our knowledge, data regarding the relationship between

ongoing HBV replication and liver related mortality in the West

are limited to patients who have already progressed to

cirrhosis.5–7 Studies conducted in tertiary care centres in

Europe among patients with compensated cirrhosis showed

that those with persistently high levels of HBV replication, as

indicated by HBeAg positivity, have increased liver related death rates, of about 2-fold, as compared to those who clear HBeAg with biochemical remission.5 Early studies on the natural history of chronic hepatitis B in Caucasian patients do not provide information whether HBeAg seroconversion leads to a decreased risk of liver-related adverse events and mortality, due

to the too short follow-up period of 5 years at most8 17 18

A prospective population-based cohort study in HBsAg positive Chinese patients has reported that high viral load at baseline (>105 copies/ml) is significantly associated with increased mortality from HCC and chronic liver disease over a 11-year period.19

However, these findings should be considered with caution because only baseline measures of HBV DNA level were tested, which are poorly related with the levels of HBV replication and disease activity during the whole follow-up of an individual Only one prospective study from Asia monitored serum HBV DNA over time and showed that patients with sustained elevation of HBV DNA had the highest HCC risk,20

but data on the risk of liver-related mortality were not reported This is the first long-term longitudinal study in untreated adult Caucasian patients with chronic hepatitis B (89% without cirrhosis at diagnosis) showing that ongoing high level of HBV replication during follow-up is associated with significantly increased risk of liver-related mortality In our study the inactive carrier was defined by sustained normal ALT and HBV DNA persistently negative by non-PCR assays, thus including a heterogeneous group of patients who may have up

to 1.46105 copies/ml of HBV DNA Nevertheless we showed that HBeAg negative patients with sustained normal ALT and HBV DNA levels of less than 1.46105 copies/ml have a very good prognosis with long life expectancy Our data suggest that recent findings from cohort studies in Chinese subjects in their 40s with perinatally or early childhood acquired HBV infection (85% of cohort HBeAg negative, 94% with normal ALT) that serum HBV DNA levels above 104copies/ml increase the risk for cirrhosis and HCC regardless of serum ALT levels and HBeAg status20 21 can not be directly referred to Caucasian patients

Figure 3 Flow diagram of the clinical outcome of 40 inactive carriers

HCC, hepatocellular carcinoma.aThree patients had cirrhosis at

enrolment and four developed cirrhosis before HBeAg seroclearance

Figure 4 Flow diagram of the clinical

outcome of 21 patients with active

hepatitis after HBeAg seroclearance

NAFLD, non-alcoholic fatty liver disease;

NASH, non-alcoholic steatohepatitis;

HCV, hepatitis C virus; HDV, hepatitis

delta virus; OLT, orthotopic liver

transplantation; HCC, hepatocellular

carcinoma.aCirrhosis at enrolment.bTwo

of the four patients developed cirrhosis

after HBeAg seroclearance.cFour of the

five patients developed cirrhosis after

HBeAg seroclearance.dResponder to

pegylated interferon and ribavirin

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Differences in long-term outcomes between Asian and

Caucasian patients should be taken into account in current

recommendations for management and treatment of chronic

hepatitis B.1 22

Notably, our data are in accordance with a recent longitudinal

study of 91 HBeAg positive Italian children showing that 81

(95%) of 85 children without cirrhosis at enrolment and who

underwent spontaneous HBeAg seroconversion remained

inac-tive carriers after 29 years follow-up and none died.23

However,

of the four children with cirrhosis two remained inactive

carriers and two developed HCC 9 and 16 years after HBeAg

seroconversion.23 In our study two inactive carriers who had

already developed cirrhosis during the HBeAg positive phase,

died of HCC 8 and 9 years after HBeAg seroconversion These

findings underline the clinical relevance of HBeAg

seroconver-sion with virological and biochemical remisseroconver-sion early in the

course of the liver disease before cirrhosis occurrence

In addition to the main findings on survival, this study

provides some data on the long-term rates of serological events

The incidence of HBeAg seroconversion was 18.7 per 100

person-years in the first 10 years of the follow-up and the

incidence of HBsAg seroconversion was 2.1 per 100 person-years

in accordance with previous studies from Western countries in

adults patients.1 2 18 24–26

Our study shows that 34% of patients developed active

hepatitis despite HBeAg seroconversion: nine patients (15%)

showed HBeAg negative hepatitis and one (1%) showed HBeAg

reversion The patients who developed HBeAg negative

hepa-titis had a significantly higher prevalence of cirrhosis at the time

of HBeAg seroconversion as compared with inactive carriers

These results are in agreement with an Asian study by Hsu

et al,27

reporting that 33% of patients with spontaneous HBeAg

seroconversion experienced ALT elevation during a median

follow-up period of 8.6 years, associated with HBeAg negative

hepatitis in 24% of cases and with HBeAg reversion in 4% In

the same study, the patients with liver cirrhosis at the time of

HBeAg seroconversion had a 10-fold increased risk of developing

HBeAg negative hepatitis Overall these data support the view

that HBeAg negative chronic hepatitis represents a late phase in

the natural history of chronic HBV infection

Concurrent HCV and/or HDV infection was found in a small

proportion (13%) of our patients with active hepatitis after

HBeAg seroconversion and did not seem to affect the natural

history of chronic hepatitis B The complex viral interplay in

cases of dual or triple infection28may provide an explanation to

the clinical evidence in longitudinal studies that HBV/HDV

co-infection is not always associated with a more severe disease

than HBV infection alone6 and that HBV/HCV concurrent

infection is not always associated with a higher risk of HCC

than in either infection alone.29

In conclusion, the present long term study in Caucasian

patients with chronic hepatitis B shows that most patients

become inactive carriers after spontaneous HBeAg

seroconver-sion and this condition confers survival benefit with regard to

liver disease mortality, particularly if present before the onset of

cirrhosis HBeAg reversion or progression to HBeAg negative

chronic hepatitis occurs in a relatively small proportion of

patients following HBeAg seroconversion The risk of

liver-related mortality is strongly associated with sustained high level

of HBV replication and disease activity during follow-up,

independently of HBeAg status Older age, male gender and

cirrhosis also strongly predict disease-related mortality

Funding: This study was partially supported by the MURST 60% 2005 of the University of Verona (GF, 2005)

Competing interests: None.

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