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Correlation between mutations in the E2 region of HCV genotype 1 isolates and responsiveness to IFNα
Posters
I P/CO6/O5 I
I P/co6/o I
S I G N S A N D S Y M P T O M S O F L I V E R D I S E A S E A F T E R 10 Y E A R S O F I N F E C T I O N W I T H H E P A T I T I S C VIRUS. DATA F R O M A NATIONAL COHORT OF PATIENTS WHO ACQUIRED THEIR INFECT I O N S O N A K N O W N DATE
S E Q U E N C E H E T E R O G E N E I T Y W I T H I N I S D R A N D P e P H D REG I O N S O F H C V AS M A R K E R O F IFN R E S P O N S E
H.E. Harris, M.E.B. Ramsay. K.P. Eldridge PHLS CDSC, London, UK. It is difficult to define the frequency and rate of progression to symptomatic or end-stage liver disease following hepatitis C vires (HCV) infection because the time of infection is rarely known and the course of infection is predominantly asymptomatic. The aim of this preliminary analysis was to investigate the prevalence of clinical signs and symptoms of liver disease after 10 years of infection. The sample comprised 874 patients who acquired their infections following transfusion of contaminated blood and who were enrolled in the UK HCV National Register. Of tbese 874 patients who had been infected for an average of 10.0 years (SEM 0.1), 116 (13.3"/o)had developed clinical signs or symptoms of liver disease (see table).
Major sign/symptom
Patients (n)
Minor sign~symptom
Liver tumor Bleeding varices Variees Ascites Splenomagaly Sub Total Grand Total
2 5 4 7 18 36
Hepatomegaly Spider naevi Other
Patients (n) 61 13 6 80 116
Of the 36 patient's whose clinicians reported 'major' signs and symptoms at the time of registration, 10 had subsequently died. Death certification showed bepatocellular carcinoma to be the cause of death for 2, while 7 died of liver failure, cirrhosis or chronic HCV. The remaining patient died of another cause unrelated to HCV infection. When the 36 patients were compared to all other patients in the register (n = 838), those who had developed 'major' symptoms were more likely to be male (69.4% vs. 48.4*/0,X~ = 6.09, P = 0.014) and to have reported drinking at least 20 units of alcohol/wk (33.3% vs. 12.9%, ):.z= 12.1g, P < 0.001). Neither age at acquisition of infection, duration of infection, anti-HBc status, ethnic group, or smoking status, were significantly different between the 2 groups (P > 0.05). These preliminary analyses suggest that in our sample of trunsfuaion acquired infections, alcohol consumption and male sex were associated with progressive HCV-related disease. These factors were highly confounded with each other (P < 0.001).
I
P/CO6/O6
]
E Puig-Basagoiti. S. A m p u r d a n 6 s , M. Gim6nez-Barcons, A. Sdnchez-Fue¥o, J. Rod6s, J.C. Sdiz, J.M. Sfi.nchez-Tapias Liver Unit, Institut de Malalties Digestive, D e p a r t a m e n t de Medicina, IDIBAPS, Hospital Clinic, U. Barcelona, Spain.
Sequence heterogeneitywithin ~e ISDRof the NS5Aof HCV has been implicated in the type of response to IFN.based therapies, likely by regulation of the activity of PKR. Recently, sequence divergence among HCV genotypes between the so-called PePHD region of the E2 protein has been also implicated in susceptibi~ to IFN, I~ely by binding and inhibiting PKR. We have partially amplified by specif~ RT-PCR b~e E2 and NS5A genes from pre-treatmentserum samples of 70 palientstreated with IFN for 6 months: 64 infected wi~ HCV-lb (4 sustained responders,SR, and 60 non-responders, NR) and 6 with HCV-3a (3 SR, 3 NR). When compared with the consensussequence, Ihe numberof aa changes withinthe ISDR of HCV-lb infected patientswas higheramong SR than NR (mean 3.7-'2.7 vs 0.9±0.8,p<.0001). No such differenceswere found in the E2 region partially sequenced. (8.5±5.8 vs 5.4±2.9), nor in the specific PePHD region (0.25±0.5 vs 0.07±0.3). No differences were found in the number of aa changes in any of Ihe regions analysed in NR and SR patients infected with HCV-3a. Pre-~atment HCV-RNA level in patients infected with HCV-lb mutated ISDR strains was significantlylower than in patients infected with HCV-lb strains beadng few or no mutations on the ISDR. Therefore, viral load and n'utationswithinthe ISDR, but not ~hin the PePHD, are related to responsivenessto IFN therapy in HCV-lb infected patients.
[ /coe/o8
]
C O R R E L A T I O N B E T W E E N M U T A T I O N S IN T H E E2 R E G I O N O F HCV GENOTYPE 1 ISOLATES AND RESPONSIVENESS TO IFNa
E F F I C A C Y O F L O N G T E R M L A M I V U D I N E T H E R A P Y IN H B e A g NEGATIVE CHRONIC HEPATITIS B
T. Berg, A. M a s - M a r q u e s , M. H6hne, U. H o p f , B. W i e d e n m a n n , E. Schreier Medizinische Klinik m. S. Hepatologie, Charit6, C a m p u s Virchow-Klinikum, R o b e r t Koch-Institut, Berlin, Germany. The reasons for the lower response rates, observed in H C V genotype 1infected patients treated with IFNa as compared to those with genotype 2 or 3 infection, are still unknown. Recently, Taylor et al. (Science, July 1999) showed that H C V envelope protein E2 contains a 12 amino acid (aa) sequence identical with phosphorylation sites of the interferon inducible protein kinase P K R a n d the translation initiation factor eIF2cc, a target o f P K R (named, PKR-eIF2cc phosphorylation homology domain [PePHD]). E2 of genotype 1 isolates binds to and inhibits PKR. This interaction of E2 a n d P K R m a y be one m e c h a n i s m b y w h i c h H C V circumvents the antiviral effect of IFNa. We investigated, whether the mutational pattern o f the PePHD o f H C V genotype 1 infected patients is correlated with the response to IFNa. Methods: Parts of the E2 region (nt.2181-3269, including the PePHD) were sequenced in 87 H C V genotype 1-infected patients with chronic hepatitis C. All patients were treated with at least 3 M U IFNa tiw. 29 of the 87 patients became H C V R N A negative during treatment (virological responders), whereas the r e m a i n i n g 58 h a d virological nonresponse. Re,mRs: The PePHD was highly conserved in most patients. Mutations in this region, leading to 1 aa exchange were found in only 6 o f t h a 87 patients a n d 3 of them showed a virological response (p=n.s). A higher variability was demonstrated in the regions up- a n d downstream o f the PePHD: m e a n 3 aa exchanges (range, 0-11) as compared to the consensus sequence. However, frequency of mutations in these regions w a s not significantly different in responder a n d nonresponder patients (mean 3.7 vs. 2.7 aa exchanges, p=0.09). The PePHD is highly conserved a m o n g H C V genotype 1infected patients a n d mutations in this region did not correlate with virological response to IFNa.
E. Dim*u, G.V. Papatheodoridis, A. Laras, S.J. Hadzivannis A c a d e m i c Dept. o f Medicine, H i p p o k r a t i o n General Hospital, Athens, Greece. Approximately 2/3 of patients (pts) with HBeAg negative (-) chronic hepatitis B (CHB) treated for 6-12 runs either with interferon-ct or lamivudine (LAM) alone achieve virological remission, but most of them relapse soon after stop of therapy. The efficacy of long-term LAM therapy in pts with HBeAg(-) CHB is currently unknown. We evaluated the safety and efficacy of long-term LAM monotherapy in a group of 25 pts (22M/3F, mean age:55+ 10 yrs) with HBeAg (-) CHB. The mean duration of this ongoing therapy has been 20.3_+6.7 (12-33) runs todate. All pts had elevated ALT levels (p.I.5xULN in 3 monthly occasions), detectable serum HBV DNA and histologically established chronic hepatitis. Pts with decompensated liver disease were excluded. LFTs were evaluated montlhy and serum HBV DNA (Monitor Roche; sensitivity: 400cp/ml) every 6 runs and on every biochemical breakthrough (BT). Median baseline ALT levels were 121 (60331) IU/L and serum HBV DNA levels 33.3 (0.08-60) xl06 cp/ml. LAM was administrated in a daily dose of 150 mg and was well tolerated; no patient discontinued or decreased the dose of the drug because of side effects. No patient lost HBsAg. The rate of biochemical response (ALT
Conclusion:
98
I P/CO6/O5 I
I P/co6/o I
S I G N S A N D S Y M P T O M S O F L I V E R D I S E A S E A F T E R 10 Y E A R S O F I N F E C T I O N W I T H H E P A T I T I S C VIRUS. DATA F R O M A NATIONAL COHORT OF PATIENTS WHO ACQUIRED THEIR INFECT I O N S O N A K N O W N DATE
S E Q U E N C E H E T E R O G E N E I T Y W I T H I N I S D R A N D P e P H D REG I O N S O F H C V AS M A R K E R O F IFN R E S P O N S E
H.E. Harris, M.E.B. Ramsay. K.P. Eldridge PHLS CDSC, London, UK. It is difficult to define the frequency and rate of progression to symptomatic or end-stage liver disease following hepatitis C vires (HCV) infection because the time of infection is rarely known and the course of infection is predominantly asymptomatic. The aim of this preliminary analysis was to investigate the prevalence of clinical signs and symptoms of liver disease after 10 years of infection. The sample comprised 874 patients who acquired their infections following transfusion of contaminated blood and who were enrolled in the UK HCV National Register. Of tbese 874 patients who had been infected for an average of 10.0 years (SEM 0.1), 116 (13.3"/o)had developed clinical signs or symptoms of liver disease (see table).
Major sign/symptom
Patients (n)
Minor sign~symptom
Liver tumor Bleeding varices Variees Ascites Splenomagaly Sub Total Grand Total
2 5 4 7 18 36
Hepatomegaly Spider naevi Other
Patients (n) 61 13 6 80 116
Of the 36 patient's whose clinicians reported 'major' signs and symptoms at the time of registration, 10 had subsequently died. Death certification showed bepatocellular carcinoma to be the cause of death for 2, while 7 died of liver failure, cirrhosis or chronic HCV. The remaining patient died of another cause unrelated to HCV infection. When the 36 patients were compared to all other patients in the register (n = 838), those who had developed 'major' symptoms were more likely to be male (69.4% vs. 48.4*/0,X~ = 6.09, P = 0.014) and to have reported drinking at least 20 units of alcohol/wk (33.3% vs. 12.9%, ):.z= 12.1g, P < 0.001). Neither age at acquisition of infection, duration of infection, anti-HBc status, ethnic group, or smoking status, were significantly different between the 2 groups (P > 0.05). These preliminary analyses suggest that in our sample of trunsfuaion acquired infections, alcohol consumption and male sex were associated with progressive HCV-related disease. These factors were highly confounded with each other (P < 0.001).
I
P/CO6/O6
]
E Puig-Basagoiti. S. A m p u r d a n 6 s , M. Gim6nez-Barcons, A. Sdnchez-Fue¥o, J. Rod6s, J.C. Sdiz, J.M. Sfi.nchez-Tapias Liver Unit, Institut de Malalties Digestive, D e p a r t a m e n t de Medicina, IDIBAPS, Hospital Clinic, U. Barcelona, Spain.
Sequence heterogeneitywithin ~e ISDRof the NS5Aof HCV has been implicated in the type of response to IFN.based therapies, likely by regulation of the activity of PKR. Recently, sequence divergence among HCV genotypes between the so-called PePHD region of the E2 protein has been also implicated in susceptibi~ to IFN, I~ely by binding and inhibiting PKR. We have partially amplified by specif~ RT-PCR b~e E2 and NS5A genes from pre-treatmentserum samples of 70 palientstreated with IFN for 6 months: 64 infected wi~ HCV-lb (4 sustained responders,SR, and 60 non-responders, NR) and 6 with HCV-3a (3 SR, 3 NR). When compared with the consensussequence, Ihe numberof aa changes withinthe ISDR of HCV-lb infected patientswas higheramong SR than NR (mean 3.7-'2.7 vs 0.9±0.8,p<.0001). No such differenceswere found in the E2 region partially sequenced. (8.5±5.8 vs 5.4±2.9), nor in the specific PePHD region (0.25±0.5 vs 0.07±0.3). No differences were found in the number of aa changes in any of Ihe regions analysed in NR and SR patients infected with HCV-3a. Pre-~atment HCV-RNA level in patients infected with HCV-lb mutated ISDR strains was significantlylower than in patients infected with HCV-lb strains beadng few or no mutations on the ISDR. Therefore, viral load and n'utationswithinthe ISDR, but not ~hin the PePHD, are related to responsivenessto IFN therapy in HCV-lb infected patients.
[ /coe/o8
]
C O R R E L A T I O N B E T W E E N M U T A T I O N S IN T H E E2 R E G I O N O F HCV GENOTYPE 1 ISOLATES AND RESPONSIVENESS TO IFNa
E F F I C A C Y O F L O N G T E R M L A M I V U D I N E T H E R A P Y IN H B e A g NEGATIVE CHRONIC HEPATITIS B
T. Berg, A. M a s - M a r q u e s , M. H6hne, U. H o p f , B. W i e d e n m a n n , E. Schreier Medizinische Klinik m. S. Hepatologie, Charit6, C a m p u s Virchow-Klinikum, R o b e r t Koch-Institut, Berlin, Germany. The reasons for the lower response rates, observed in H C V genotype 1infected patients treated with IFNa as compared to those with genotype 2 or 3 infection, are still unknown. Recently, Taylor et al. (Science, July 1999) showed that H C V envelope protein E2 contains a 12 amino acid (aa) sequence identical with phosphorylation sites of the interferon inducible protein kinase P K R a n d the translation initiation factor eIF2cc, a target o f P K R (named, PKR-eIF2cc phosphorylation homology domain [PePHD]). E2 of genotype 1 isolates binds to and inhibits PKR. This interaction of E2 a n d P K R m a y be one m e c h a n i s m b y w h i c h H C V circumvents the antiviral effect of IFNa. We investigated, whether the mutational pattern o f the PePHD o f H C V genotype 1 infected patients is correlated with the response to IFNa. Methods: Parts of the E2 region (nt.2181-3269, including the PePHD) were sequenced in 87 H C V genotype 1-infected patients with chronic hepatitis C. All patients were treated with at least 3 M U IFNa tiw. 29 of the 87 patients became H C V R N A negative during treatment (virological responders), whereas the r e m a i n i n g 58 h a d virological nonresponse. Re,mRs: The PePHD was highly conserved in most patients. Mutations in this region, leading to 1 aa exchange were found in only 6 o f t h a 87 patients a n d 3 of them showed a virological response (p=n.s). A higher variability was demonstrated in the regions up- a n d downstream o f the PePHD: m e a n 3 aa exchanges (range, 0-11) as compared to the consensus sequence. However, frequency of mutations in these regions w a s not significantly different in responder a n d nonresponder patients (mean 3.7 vs. 2.7 aa exchanges, p=0.09). The PePHD is highly conserved a m o n g H C V genotype 1infected patients a n d mutations in this region did not correlate with virological response to IFNa.
E. Dim*u, G.V. Papatheodoridis, A. Laras, S.J. Hadzivannis A c a d e m i c Dept. o f Medicine, H i p p o k r a t i o n General Hospital, Athens, Greece. Approximately 2/3 of patients (pts) with HBeAg negative (-) chronic hepatitis B (CHB) treated for 6-12 runs either with interferon-ct or lamivudine (LAM) alone achieve virological remission, but most of them relapse soon after stop of therapy. The efficacy of long-term LAM therapy in pts with HBeAg(-) CHB is currently unknown. We evaluated the safety and efficacy of long-term LAM monotherapy in a group of 25 pts (22M/3F, mean age:55+ 10 yrs) with HBeAg (-) CHB. The mean duration of this ongoing therapy has been 20.3_+6.7 (12-33) runs todate. All pts had elevated ALT levels (p.I.5xULN in 3 monthly occasions), detectable serum HBV DNA and histologically established chronic hepatitis. Pts with decompensated liver disease were excluded. LFTs were evaluated montlhy and serum HBV DNA (Monitor Roche; sensitivity: 400cp/ml) every 6 runs and on every biochemical breakthrough (BT). Median baseline ALT levels were 121 (60331) IU/L and serum HBV DNA levels 33.3 (0.08-60) xl06 cp/ml. LAM was administrated in a daily dose of 150 mg and was well tolerated; no patient discontinued or decreased the dose of the drug because of side effects. No patient lost HBsAg. The rate of biochemical response (ALT
Conclusion:
98