- Email: [email protected]
Viral kinetics on treatment with pegylated (PEG 40 KD) interferon (IFN) alfa2a in patients infected with HCV subtype 1
Category 6: Viral hepatitis: clinical aspects stimulation, while substitution of recombinant IL-12 in cocultures of HBV patients recovered Th cell stimulation. Conclusions: An impaired differentiation of DC from myeloid precursor cells associated with an IL-12 secretion failure might lead to reduced Thl cell stimulation and therefore cause viral persistence in patients with chronic hepatitis B. Thus, IL-12 might be a future therapeutic agent.
' ~ - ] MUTATIONS OF PRES2 B CELL EPITOPE CLUSTER IN LIVER TRANSPLANT PATIENTS WITH RECURRENT HBV INFECTION Erica Villa. Internal Medicine, University of Modena, Italy To evaluate whether pretransplant HBV surface gene mutations could be associated with higher recurrence, preS/S region was sequenced. Methods: pretransplant sera of 14 HBV+ liver transplant (LT) patients (9 recurrent and 5 not) were studied. None had lamivudine before LT, all underwent HIBG i.v. HBV DNA was amplified by PCR with preS/S primers and directly sequenced. Results: 77 mutations/l167 (6.6%) bases sequenced were found; 39 gave rise to an aa change. Five aa changes were present in almost all patients independently from recurrence (preS2: codons 144 (11/11); 149 or 150 (9/11); S: 125 (10/11), 127 (8/11). Missense mutations were significantly more present in recurrent patients (p = 0.009, Mann-Whitney) and clustered preferentially in the preS2 region (preS2: rec. vs. non rec: p = 0.007). Most preS2 mutations (9/14) unique to recurrent pts were localised between codons 8 and 37 (potential preS2 B cell epitope). In 1 recurrent p. onset of a TAG codon from C-to-T mutation prematurely terminated preS1 expression, in another, M109V mutation abolished expression of preS2 protein. Conclusions: 1. Profound sequence divergence from ayw sequence and changes of "a" determinant were present before LT in all pts irrespective of recurrence after LT; 2. In recurrent pts, missense mutations clustered in preS2 region, especially in putative B cell epitope. In conclusion, HBV recurrence could have been facilitated by the alteration of B cell epitopes of S but especially of preS2 protein which, leading to conformational changes, despite very low levels of HBV replication before LT, may have determined decreased efficacy or even ineffectiveness of HBIG.
7 DETECTION OF HEPATITIS C VIRUS RNA IN SALIVARY GLANDS BY IN SITU HYBRIDIZATION
131
chronic sialadenitits. However, its implication in the pathogenesis of these diseases deserves future research.
-] NATURAL KILLER (NK) CELL CYTOTOXlCITY IN CHRONIC HCV INFECTION. RELATIONSHIP WITH CLINICAL AND VIROLOGICAL FEATURES AND RESPONSE TO INTERFERON (IFN) G. Castiglione, A. Guglietta 1, F. Lirussi. Dept. of Medical and Surgical
Sciences, University of Padova, Italy; 1 Grupo Ferrer International, Barcelona, Spain The role of NK cells in HCV-related chronic liver disease is largely unknown. The aim of the present study was twofold: 1) to study peripheral NK cell cytotoxicity in relation to liver function and structure as well as HCV genotypes (lb vs non-lb) and viremia; 2) to investigate whether NK cell cytotoxicity detected before a twelve-month IFN-a treatment could be a predictor of response. Fifteen patients with biopsy-proven HCV-related chronic liver disease and 23 matched healthy volunteers were studied. Patients were investigated for conventional liver function tests, histology (grading and staging), quantitative HCV-RNA and genttypes. NK cell cytotoxic activity was evaluated by means of a newly developed ELISA assay based on the assessment of NK cells-induced apoptosis of target cells (K562). NK cell cytotoxicity for an effector:target cell ratio of 50:1 was almost half In HCV-positive patients compared to controls (26.4 -4- 4.3 SEM vs 47.1 d: 4.7 times x basal activity; p = 0.0026). NK cell activity was inversely related to age both in the patients (r = -0.32; p = 0.0169) and the controls (r = -0.58; p = 0.0039) but did not correlate with ALT levels, histopathologic lesions, viremia and genotypes (p = n.s.). Moreover, no significant differences were found between responders (n = 6) and non-responders (n = 7) in respect to NK cell cytotoxicity. In conclusion, NK cell cytotoxic activity is markedly impaired in chronic HCV infection, based on an original assay devoid of the risk associated with the use of radioactive material. Such activity seems independent from viral and host characteristics except age and does not help predict the outcome of IFN treatment.
P06
Category 6: Viral hepatitis: clinical aspects
E. Rodriguez-Inigo, N. Ortiz-Movilla, J. Bartolome, M. Pardo, V. Carreno. Fundacion Estudio Hepatitis Virales and Instituto de
Hepatologia, Hospital Pardo de Aravaca, Spain HCV-RNA has been detected by PCR in saliva and salivary glands from patients with sialadenitis and Sjtgren's syndrome. However, morphological evidence of HCV replication in salivary glands is needed to support a role for HCV in causing these diseases. We have analyzed by in situ hybridization the presence of HCV-RNA of sense and antisense polarity in salivary gland biopsies from 13 patients with chronic sialadenitis and 6 with Sjtgren's syndrome. Eight of them (1 Sjtgren's syndrome and 7 sialadenitis) were anti-HCV positive and had serum HCV-RNA. HCV-RNA of both polarities was detected in the cytoplasm of epithelial cells of the salivary glands from all the anti-HCV positive patients but in none of the anti-HCV negative. Positive cells to the HCV probe of positive polarity showed positive signals using the probe of negative polarity, demonstrating the viral replication in these cells. The percentage of HCV positive cells ranged from 25% to 48.8%. No differences in the lesions between patients with and without HCV in the salivary glands were found. HCV infected cells did not show any morphological difference with respect to uninfected ceils. Thus, HCV infects and replicates in salivary glands of patients with Sjt~gren's syndrome or
[-~
VIRAL KINETICS ON TREATMENT WITH PEGYLATED (PEG 40 KD) INTERFERON (IFN) ALFA2A IN PATIENTS INFECTED WITH HCV SUBTYPE 1
P. Ferenci, Rudolf Stauber, Franz Hackl, Wolfgang Jessner, Karin Hegenbarth, Petra Munda-Steindl, Harald Kessler. Internal
Medicine IV, Univ. of Vienna, Austria Viral kinetics is useful to study antiviral therapy in chronic hepatitis C CHC). Viral decline is described in two phases, a rapid first and a second slower one and was determined following PEG (40 kd)-IFNalfa2a (Roche, CH). Methods: 20 IFN-nai've CHC-patients infected by HCV-la/b (m: 12, f: 8; median age: 44 years [range: 30--64]; ALT: 50 IU/L [28-214]; fibrosis stage 1-2: n = 13; 3: n = 4; 4: n = 3) were studied. First, the viral load was measured 24 h after 9 MU IFNalfa2a (Roferon, Roche, CH) (TO and T1). One week later 180/zg PEG (40 kd)-IFN alfa2a/week for 2 weeks (DO to D13) was given. Thereafter (D14) 1000-1200 mg ribavirin/d was added for a total treatment of 50 weeks. Viral load was determined by
132
Poster Sessions
the Cobas Amplicor Monitor HCV 2.0 (Roche Diagnostics, Branchburg, NY) on TO, T1, DO, D1, D4, D7, and D13. Results: Viral load was similar on TO (5.6 [5.02-6] log IU/mL) and DO (5.5 [5.15-5.95]). In the 6 likely IFN-nonresponders (defined by T1 < 70% TO) HCV did not decrease on PEG (40 kd)-IFNalfa2a. In contrast, in 10 of the 14 IFN-sensitive patients (HCV TI: -1.07 [0.65-1.67] log IU/mL) viral load declined markedly on D13 (-1.44 4- 2.1 [SD]). However, as compared with T1, the median decline was less pronounced at D1, D4, and D7 (0.41; 0.63; 0.62; p < 0.01). Conclusion: Initial viral elimination on PEG (40 kd)-IFNalfa2a is different from previously described experience on daily high-dose standard IFN. The initial decline is less pronounced. Nevertheless, after the second dose of PEG (40 kd)-IFNalfa2a IFN-sensitive patients have a more than 90% decline of the initial viral load.
-~
INDUCTION INTERFERON ALFA-2A MIGHT IMPROVE THE OUTCOME OF COMPENSATED HEPATITIS C VIRUS (HCV)-RELATED CIRRHOSIS
R. Planas, LC. Quer, J. Enri-Quez, J.M. Bah'era, B. Dalmau, T. Casanovas, J.M. Viver, M. Tortes, J. Boadas, R. Solagrave. Dept. of
Gastroenterology, Hospital Universitari Germans Trias 1 Pujol, Badalona, Spain Although standard dose IFN is successful in only 5% of patients with HCV-related cirrhosis, it has been suggested that might decrease the risk of complications or the incidence of hepatocellular carcinoma. Based on HCV kinetics, daily IFN may improve response rates. Aims: 1) To assess the efficacy and tolerability of induction IFN therapy in compensated HCV-related cirrhosis. 2) To evaluate the impact of treatment on the probability of developing decompensation or hepatocellular carcinoma during the posttrial 3-year follow-up. Patients: Forty cirrhotic patients were randomized to receive (Group I: 19) or not (Group II: 21) treatment with 1FN (4.5 MU/daily for 24 weeks, followed by 4.5 MU/48 h, only if ALT was within normal values). Results: Dose reduction and discontinuation for adverse events was required in 11 (58%) and 6 (31.5%) cases. End-of-treatment response was not observed in any of the 21 controls, but was observed in 4 of the 19 (21%) treated patients (p = 0.04); sustained response was obtained in only 2 (10.5%) treated patients. The 3-year probability of developing any of the following: ascites, hepatocellular carcinoma, transplantation or death, was lower in Group I than in Group II (6% vs. 27%; p = 0.05). Conclusions: Although induction IFN therapy is associated with frequent side effects and poor sustained response in compensated HCVrelated cirrhosis, it might improve the outcome of the patients.
~2]
PREDICTORS OF POSITIVE HEPATITIS C SEROLOGY IN INTRAVENOUS DRUG USERS
M. Backmund, K. Meyer, M. von Zielonka, D. Eichenlaub. Addiction
Medicine, General Hospital Schwabing, Munich, Germany Background: Hepatitis C Virus (HCV) infection is the most common disease among intravenous drug users (IDU). Patients and Method: All patients admitting to detoxification unit 1991-1997 and meeting ICD-10 diagnosis of opioid dependency were tested for anti-HCV serology. Results: 1047 patients were included in the study. 62.8% of the IDU's were anti-HCV positive. Increasing age (p < 0.01), living with a significant other drug user (p < 0.01), history of therapy (p < 0.01), history of imprisonment (p < 0.01), history of emergency treatment (p < 0.01), additional daily consumption of benzodiazepines (p < 0.05) or alcohol (p < 0.01), frequency of injecting heroin (p < 0.05) and type of opioid dependency (p < 0.01) were significant associated as individual factors with anti-HCV serology. Using multiple logistic regression we found
that age (p = 0.0000), living with a significant other drug user (p = 0.0111), history of therapy (p = 0.0000), history of imprisonment (p = 0.0174), history of emergency treatment (p = 0.0003) and additional daily consumption of alcohol (p = 0.0016) remained independently associated with positive anti-HCV serology. Conclusions: These data support the need for early prevention strategies, namely, education of the teachers in schools and further training of the counsellors informing the IDU's of what they can do to minimise the risk of becoming infected or of transmitting infectious agents to others.
~0-] SERUM HBV DNA LEVEL AND LIVER HISTOLOGY IN HEALTHY HBSAG CARRIERS Michelle Martinot-Peignoux, Raoudha Akremi, Magali Colombat, Veronique Le Breton, Nathalie Boyer, Claude Degott, Patrick Marcellin. France The NIH Consensus workshop on HBV (September 2000) revisited the definition of Healthy HBsAg carders. The new definition Inactive HBsAg carriers includes quantification of serum HBV DNA. However, serum HBV DNA cutoff allowing identifying this group of subjects remains to be determined. Patients: 73 patients, HBsAg positive/HBeAg negative with persistently normal ALT (*40 UI/L) and undetectable serum HBV DNA with standard assays, were followed-up 3 4- 2.8 years (0.5 to 11), 55 had a liver biopsy. Methods: HBV DNA was quantified with PCR (Cobas Amplicor HBV MONITOR, Roche) (sensitivity < 102 copies/ml), liver histology assessed using Ishak scoring system (0 to 24). Results: Median HBV DNA load was 1.34 x 103 copies/ml (range <102 to 2.1 x 105), 12% of the patients had HBV DNA level < 102 and 5% > 105 copies/ml. HBV DNA level was significantly higher when a threshold of *30 UIFL was considered for ALT (p = 0.002). Histologic lesions were mild in all cases, mean Ishak score was 3.5 4- 1.6 (range 0 to 6), and no patient had a fibrosis score above 3. During the follow-up 3 patients lost HBsAg (all HBV DNA < 102), 3 patients experienced a mild and transient increase in ALT. Among the 11 patients repeatedly tested for HBV DNA 10 remained stable. Additional data on serum HBV DNA and liver histologic outcome will be presented. Conclusion: In our study, using the NIH new definition for "Inactive HBsAg carriers", 95% of the patients had HBV DNA level < 105 copies/ml and remained stable. Histologic lesions were mild in all cases.
~
LONG-TERM LAMIVUDINE TREATMENT IN ANTI-HBE POSITIVE CHRONIC HEPATITIS B: AN INTERIM ANALYSIS
T. Santantonio, E. Sinisi, M. Mazzola, M. Insalata, A. Guastadisegni, G. Pastore. Clinica Malattie lnfettive, Universittidi Bari, Bari, Italy We evaluated the efficacy of long-term LAM at 100 mg once dally in 36 chronic anti-HBe positive hepatitis B pts (30 males and 6 females, mean age 49 yrs), and the incidence and clinical implications of YMDD mutants. All pts received LAM for one year and 23 were treated for at least 18 mos; undetectable HBV DNA by molecular hybridization and nomal ALT were observed in all but two pts. Eleven pts showed virological and biochemical breakthrough after at least 6 mos of therapy and all had mutations in the YMDD motif (M552V/L528M, M552I/L528M, M552I). In 5/11 pts, the YMDD mutant emergence was associated with ALT levels > 10 times the upper normal value, whereas in the remaining six, ALT levels did not exceed the baseline. LAM was discontinued in these 11 pts after a mean period of 6 mos because of persistent viremia and increased ALT, or because the same fluctuating biochemical and virological profile was observed as that before treatment. In 5 pts, a liver biopsy was performed during treatment after breakthrough. A more than two-point increase or reduction in histological activity index was found in 3 and 2 pts, respectively. Our data confirm the antiviral efficacy
' ~ - ] MUTATIONS OF PRES2 B CELL EPITOPE CLUSTER IN LIVER TRANSPLANT PATIENTS WITH RECURRENT HBV INFECTION Erica Villa. Internal Medicine, University of Modena, Italy To evaluate whether pretransplant HBV surface gene mutations could be associated with higher recurrence, preS/S region was sequenced. Methods: pretransplant sera of 14 HBV+ liver transplant (LT) patients (9 recurrent and 5 not) were studied. None had lamivudine before LT, all underwent HIBG i.v. HBV DNA was amplified by PCR with preS/S primers and directly sequenced. Results: 77 mutations/l167 (6.6%) bases sequenced were found; 39 gave rise to an aa change. Five aa changes were present in almost all patients independently from recurrence (preS2: codons 144 (11/11); 149 or 150 (9/11); S: 125 (10/11), 127 (8/11). Missense mutations were significantly more present in recurrent patients (p = 0.009, Mann-Whitney) and clustered preferentially in the preS2 region (preS2: rec. vs. non rec: p = 0.007). Most preS2 mutations (9/14) unique to recurrent pts were localised between codons 8 and 37 (potential preS2 B cell epitope). In 1 recurrent p. onset of a TAG codon from C-to-T mutation prematurely terminated preS1 expression, in another, M109V mutation abolished expression of preS2 protein. Conclusions: 1. Profound sequence divergence from ayw sequence and changes of "a" determinant were present before LT in all pts irrespective of recurrence after LT; 2. In recurrent pts, missense mutations clustered in preS2 region, especially in putative B cell epitope. In conclusion, HBV recurrence could have been facilitated by the alteration of B cell epitopes of S but especially of preS2 protein which, leading to conformational changes, despite very low levels of HBV replication before LT, may have determined decreased efficacy or even ineffectiveness of HBIG.
7 DETECTION OF HEPATITIS C VIRUS RNA IN SALIVARY GLANDS BY IN SITU HYBRIDIZATION
131
chronic sialadenitits. However, its implication in the pathogenesis of these diseases deserves future research.
-] NATURAL KILLER (NK) CELL CYTOTOXlCITY IN CHRONIC HCV INFECTION. RELATIONSHIP WITH CLINICAL AND VIROLOGICAL FEATURES AND RESPONSE TO INTERFERON (IFN) G. Castiglione, A. Guglietta 1, F. Lirussi. Dept. of Medical and Surgical
Sciences, University of Padova, Italy; 1 Grupo Ferrer International, Barcelona, Spain The role of NK cells in HCV-related chronic liver disease is largely unknown. The aim of the present study was twofold: 1) to study peripheral NK cell cytotoxicity in relation to liver function and structure as well as HCV genotypes (lb vs non-lb) and viremia; 2) to investigate whether NK cell cytotoxicity detected before a twelve-month IFN-a treatment could be a predictor of response. Fifteen patients with biopsy-proven HCV-related chronic liver disease and 23 matched healthy volunteers were studied. Patients were investigated for conventional liver function tests, histology (grading and staging), quantitative HCV-RNA and genttypes. NK cell cytotoxic activity was evaluated by means of a newly developed ELISA assay based on the assessment of NK cells-induced apoptosis of target cells (K562). NK cell cytotoxicity for an effector:target cell ratio of 50:1 was almost half In HCV-positive patients compared to controls (26.4 -4- 4.3 SEM vs 47.1 d: 4.7 times x basal activity; p = 0.0026). NK cell activity was inversely related to age both in the patients (r = -0.32; p = 0.0169) and the controls (r = -0.58; p = 0.0039) but did not correlate with ALT levels, histopathologic lesions, viremia and genotypes (p = n.s.). Moreover, no significant differences were found between responders (n = 6) and non-responders (n = 7) in respect to NK cell cytotoxicity. In conclusion, NK cell cytotoxic activity is markedly impaired in chronic HCV infection, based on an original assay devoid of the risk associated with the use of radioactive material. Such activity seems independent from viral and host characteristics except age and does not help predict the outcome of IFN treatment.
P06
Category 6: Viral hepatitis: clinical aspects
E. Rodriguez-Inigo, N. Ortiz-Movilla, J. Bartolome, M. Pardo, V. Carreno. Fundacion Estudio Hepatitis Virales and Instituto de
Hepatologia, Hospital Pardo de Aravaca, Spain HCV-RNA has been detected by PCR in saliva and salivary glands from patients with sialadenitis and Sjtgren's syndrome. However, morphological evidence of HCV replication in salivary glands is needed to support a role for HCV in causing these diseases. We have analyzed by in situ hybridization the presence of HCV-RNA of sense and antisense polarity in salivary gland biopsies from 13 patients with chronic sialadenitis and 6 with Sjtgren's syndrome. Eight of them (1 Sjtgren's syndrome and 7 sialadenitis) were anti-HCV positive and had serum HCV-RNA. HCV-RNA of both polarities was detected in the cytoplasm of epithelial cells of the salivary glands from all the anti-HCV positive patients but in none of the anti-HCV negative. Positive cells to the HCV probe of positive polarity showed positive signals using the probe of negative polarity, demonstrating the viral replication in these cells. The percentage of HCV positive cells ranged from 25% to 48.8%. No differences in the lesions between patients with and without HCV in the salivary glands were found. HCV infected cells did not show any morphological difference with respect to uninfected ceils. Thus, HCV infects and replicates in salivary glands of patients with Sjt~gren's syndrome or
[-~
VIRAL KINETICS ON TREATMENT WITH PEGYLATED (PEG 40 KD) INTERFERON (IFN) ALFA2A IN PATIENTS INFECTED WITH HCV SUBTYPE 1
P. Ferenci, Rudolf Stauber, Franz Hackl, Wolfgang Jessner, Karin Hegenbarth, Petra Munda-Steindl, Harald Kessler. Internal
Medicine IV, Univ. of Vienna, Austria Viral kinetics is useful to study antiviral therapy in chronic hepatitis C CHC). Viral decline is described in two phases, a rapid first and a second slower one and was determined following PEG (40 kd)-IFNalfa2a (Roche, CH). Methods: 20 IFN-nai've CHC-patients infected by HCV-la/b (m: 12, f: 8; median age: 44 years [range: 30--64]; ALT: 50 IU/L [28-214]; fibrosis stage 1-2: n = 13; 3: n = 4; 4: n = 3) were studied. First, the viral load was measured 24 h after 9 MU IFNalfa2a (Roferon, Roche, CH) (TO and T1). One week later 180/zg PEG (40 kd)-IFN alfa2a/week for 2 weeks (DO to D13) was given. Thereafter (D14) 1000-1200 mg ribavirin/d was added for a total treatment of 50 weeks. Viral load was determined by
132
Poster Sessions
the Cobas Amplicor Monitor HCV 2.0 (Roche Diagnostics, Branchburg, NY) on TO, T1, DO, D1, D4, D7, and D13. Results: Viral load was similar on TO (5.6 [5.02-6] log IU/mL) and DO (5.5 [5.15-5.95]). In the 6 likely IFN-nonresponders (defined by T1 < 70% TO) HCV did not decrease on PEG (40 kd)-IFNalfa2a. In contrast, in 10 of the 14 IFN-sensitive patients (HCV TI: -1.07 [0.65-1.67] log IU/mL) viral load declined markedly on D13 (-1.44 4- 2.1 [SD]). However, as compared with T1, the median decline was less pronounced at D1, D4, and D7 (0.41; 0.63; 0.62; p < 0.01). Conclusion: Initial viral elimination on PEG (40 kd)-IFNalfa2a is different from previously described experience on daily high-dose standard IFN. The initial decline is less pronounced. Nevertheless, after the second dose of PEG (40 kd)-IFNalfa2a IFN-sensitive patients have a more than 90% decline of the initial viral load.
-~
INDUCTION INTERFERON ALFA-2A MIGHT IMPROVE THE OUTCOME OF COMPENSATED HEPATITIS C VIRUS (HCV)-RELATED CIRRHOSIS
R. Planas, LC. Quer, J. Enri-Quez, J.M. Bah'era, B. Dalmau, T. Casanovas, J.M. Viver, M. Tortes, J. Boadas, R. Solagrave. Dept. of
Gastroenterology, Hospital Universitari Germans Trias 1 Pujol, Badalona, Spain Although standard dose IFN is successful in only 5% of patients with HCV-related cirrhosis, it has been suggested that might decrease the risk of complications or the incidence of hepatocellular carcinoma. Based on HCV kinetics, daily IFN may improve response rates. Aims: 1) To assess the efficacy and tolerability of induction IFN therapy in compensated HCV-related cirrhosis. 2) To evaluate the impact of treatment on the probability of developing decompensation or hepatocellular carcinoma during the posttrial 3-year follow-up. Patients: Forty cirrhotic patients were randomized to receive (Group I: 19) or not (Group II: 21) treatment with 1FN (4.5 MU/daily for 24 weeks, followed by 4.5 MU/48 h, only if ALT was within normal values). Results: Dose reduction and discontinuation for adverse events was required in 11 (58%) and 6 (31.5%) cases. End-of-treatment response was not observed in any of the 21 controls, but was observed in 4 of the 19 (21%) treated patients (p = 0.04); sustained response was obtained in only 2 (10.5%) treated patients. The 3-year probability of developing any of the following: ascites, hepatocellular carcinoma, transplantation or death, was lower in Group I than in Group II (6% vs. 27%; p = 0.05). Conclusions: Although induction IFN therapy is associated with frequent side effects and poor sustained response in compensated HCVrelated cirrhosis, it might improve the outcome of the patients.
~2]
PREDICTORS OF POSITIVE HEPATITIS C SEROLOGY IN INTRAVENOUS DRUG USERS
M. Backmund, K. Meyer, M. von Zielonka, D. Eichenlaub. Addiction
Medicine, General Hospital Schwabing, Munich, Germany Background: Hepatitis C Virus (HCV) infection is the most common disease among intravenous drug users (IDU). Patients and Method: All patients admitting to detoxification unit 1991-1997 and meeting ICD-10 diagnosis of opioid dependency were tested for anti-HCV serology. Results: 1047 patients were included in the study. 62.8% of the IDU's were anti-HCV positive. Increasing age (p < 0.01), living with a significant other drug user (p < 0.01), history of therapy (p < 0.01), history of imprisonment (p < 0.01), history of emergency treatment (p < 0.01), additional daily consumption of benzodiazepines (p < 0.05) or alcohol (p < 0.01), frequency of injecting heroin (p < 0.05) and type of opioid dependency (p < 0.01) were significant associated as individual factors with anti-HCV serology. Using multiple logistic regression we found
that age (p = 0.0000), living with a significant other drug user (p = 0.0111), history of therapy (p = 0.0000), history of imprisonment (p = 0.0174), history of emergency treatment (p = 0.0003) and additional daily consumption of alcohol (p = 0.0016) remained independently associated with positive anti-HCV serology. Conclusions: These data support the need for early prevention strategies, namely, education of the teachers in schools and further training of the counsellors informing the IDU's of what they can do to minimise the risk of becoming infected or of transmitting infectious agents to others.
~0-] SERUM HBV DNA LEVEL AND LIVER HISTOLOGY IN HEALTHY HBSAG CARRIERS Michelle Martinot-Peignoux, Raoudha Akremi, Magali Colombat, Veronique Le Breton, Nathalie Boyer, Claude Degott, Patrick Marcellin. France The NIH Consensus workshop on HBV (September 2000) revisited the definition of Healthy HBsAg carders. The new definition Inactive HBsAg carriers includes quantification of serum HBV DNA. However, serum HBV DNA cutoff allowing identifying this group of subjects remains to be determined. Patients: 73 patients, HBsAg positive/HBeAg negative with persistently normal ALT (*40 UI/L) and undetectable serum HBV DNA with standard assays, were followed-up 3 4- 2.8 years (0.5 to 11), 55 had a liver biopsy. Methods: HBV DNA was quantified with PCR (Cobas Amplicor HBV MONITOR, Roche) (sensitivity < 102 copies/ml), liver histology assessed using Ishak scoring system (0 to 24). Results: Median HBV DNA load was 1.34 x 103 copies/ml (range <102 to 2.1 x 105), 12% of the patients had HBV DNA level < 102 and 5% > 105 copies/ml. HBV DNA level was significantly higher when a threshold of *30 UIFL was considered for ALT (p = 0.002). Histologic lesions were mild in all cases, mean Ishak score was 3.5 4- 1.6 (range 0 to 6), and no patient had a fibrosis score above 3. During the follow-up 3 patients lost HBsAg (all HBV DNA < 102), 3 patients experienced a mild and transient increase in ALT. Among the 11 patients repeatedly tested for HBV DNA 10 remained stable. Additional data on serum HBV DNA and liver histologic outcome will be presented. Conclusion: In our study, using the NIH new definition for "Inactive HBsAg carriers", 95% of the patients had HBV DNA level < 105 copies/ml and remained stable. Histologic lesions were mild in all cases.
~
LONG-TERM LAMIVUDINE TREATMENT IN ANTI-HBE POSITIVE CHRONIC HEPATITIS B: AN INTERIM ANALYSIS
T. Santantonio, E. Sinisi, M. Mazzola, M. Insalata, A. Guastadisegni, G. Pastore. Clinica Malattie lnfettive, Universittidi Bari, Bari, Italy We evaluated the efficacy of long-term LAM at 100 mg once dally in 36 chronic anti-HBe positive hepatitis B pts (30 males and 6 females, mean age 49 yrs), and the incidence and clinical implications of YMDD mutants. All pts received LAM for one year and 23 were treated for at least 18 mos; undetectable HBV DNA by molecular hybridization and nomal ALT were observed in all but two pts. Eleven pts showed virological and biochemical breakthrough after at least 6 mos of therapy and all had mutations in the YMDD motif (M552V/L528M, M552I/L528M, M552I). In 5/11 pts, the YMDD mutant emergence was associated with ALT levels > 10 times the upper normal value, whereas in the remaining six, ALT levels did not exceed the baseline. LAM was discontinued in these 11 pts after a mean period of 6 mos because of persistent viremia and increased ALT, or because the same fluctuating biochemical and virological profile was observed as that before treatment. In 5 pts, a liver biopsy was performed during treatment after breakthrough. A more than two-point increase or reduction in histological activity index was found in 3 and 2 pts, respectively. Our data confirm the antiviral efficacy