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HCV genotypes in patients with chronic hepatitis C and silent HCV infection after sustained biochemical response to interferon
$24
Posters 1
L, I P1 C1/05 I
[ P1 C1/07 I
IMMUNE MECHANISMSDURING INTERFERON.ALPHA (IFN) INDUCED VIRUSELIMINATIONIN CHRONICHBV AND HCV INFECTION:ANALYSIS OF VIRUS-SPECIFICCD4+ T LYMPHOCY'rES
HCV GENOTYPES IN PATIENTS WITH CHRONIC HEPATITIS C AND SILENT HCV INFECTION AFTER SUSTAINED BIOCHEMICAL RESPONSE TO INTERFERON
HM Diepolder, MC Jung, RM Hoffinao., R Zaehoval, FM Zwiebel, C Korherr, G Paumgarmer, G Riethmiiller, and GR Pape. luslitute of hnmunology and Dept. of Internal Medicine II, Klinikum GroBhadern. University of Munich, Germany. Virusspecific CD4+ T-I~mphoeytes are believed to be the mainstay of the amiviral immune response in several viral diseases, including influenza and acute hepatitis B. While [FN has been shown to be an effective treatment for a subgroup of patients with chronic hepatitis B (criB) and chronic hepatitis C (cHC). the role of virusspecific CIM+ T cells in viral elimination during IFN therapy is unknown. We serially studied the virusspecifie proliferative response of peripheral CD4+ T lymphocytes (CD4+T-R) to recombinant viral proteins in 10 patients with crib and 27 patients will1 cHC who completed a course of IFN therapy. None of tile patients with criB had a CD4+T-R to hepalitis B virus (HBV) core protein before IFN therapy. However. during IFN therapy those patients who developed a "flare" of liver inflammation and seroeonverted to anti-HBe transiently displayed a vigorous CD4+T-R Io HBV core protein. These results arc in agreement with studies that show an association of s CD4+T-R to HBe/core protein with viral elimination in acute hepatitis B as well" as spontaneous viral clearance in criB. In contrast, 13/27 patients with cHC showed a CD4+T-R to hepatitis C virus (HCV) core protein, NS3. NS4. or NS5 already before IFN therapy and sustained responders (SR) were confined to this group. A new CD4+T-R to HCV proteins during IFN therapy was typically present in transient responders during the intervall of response but was frequently lost again after relapse. Neither permanent or transient loss of HCV-RNA was preceded by an increased inflammatory activity. Our data suggest that a virus-specific CD4+T-R seems to be important for viral eliminiation during IFN therapy in both crib and cHC. However, while in criB an HBV-specific CD4+T-R is induced by IFN and accompanied by viral elimination, in eric only those patients with a high a priori CD4+T-R are apparently able to clear the infection. A detailed analysis of IFN induced immune mechanismsin cH'B and cHC may be a key for Ihe development of new rational therapies.
P.Pontisso t L.Chemello t C.Casarin t HG.Ruvoletto, H.Gerotto t A.Alberti. Clinica Hedica 2, University of Padova (Italy) Interferon treatment in patients with chronic hepatitis type C leads to complete response, in terms of biochemical remission, in about 25Z of the patients. ALT normalization is not invariably associated with viral clearance and serum HCV-RNA can persist after therapy in individual cases. To evaluate whether this behaviour is related to HCV genotype, 316 patients with chronic hepatitis C treated with interferon (3 to 9 HU/tiw for 6 to 12 months) and followed up for at least 12 months after therapy were studied. For genotyping, hybridization of the 5'UTR amplified products with probes specific for different HCV genotypes was used. Among 253 viremic patients, 46~ were infected by HCV-I (21Z la and 79Z Ib), 42Z by HCV-2, ii~ by HCV-3 and I~ by HCV-4. Long term response was observed in 85 patients, representing 9Z of HCV-I, 52~ of HCV-2 and 59Z of HCV-3 infected patients. Among these sustained responders with normal ALT, serum HCV-RNA persisted 6 to 12 months after therapy in 20Z of patients with HCV-I and in 45Z of those with HCV-2, while it was never found in those with HCV-3. Viral load was invariably decreased compared to basal levels. In conclusion, HCV-I and particularly HCV-2 but not HCV-3 are associated with clinically silent HCV infection after sustained biochemical response to interferon therapy.
I P1 c1/061
I Pl Cl/08
LONG-TERM RESPONSE TO INTERFERON THERAPY IN HEPATITIS B VIRUS (HBV) PRE-C MUTANTS ASSOCIATED CHRONIC HEPATITIS.
HCV VIRAEMIA, VIRAL GENOTYPES AND RESPONSE TO IFN IN PATIENTS WITH CHRONIC HEPATITIS C FROM SICILY.
T. Santantonio 1,2, M.Milella 1, L.Monno1' T.lacovazzi 1' G.Pastorel. H.Will 2 1Clinic of Infectious Diseases, Universita' dl Bad, Italy. 2Heinrich-Pette-lnstitut, University of Hamburg, Germany Objective. To evaluate the long-term therapeutic efficacy of alpha interferon (alFN) in chronic anti-HBe+ hepatitis B due to HBV pre-C mutants containing mutations in the pre-C region which prevent precora and e-antigen expression. Methods. 42 patients Cots) with anti-HBe/HBV-DNA+ chronic hepatitis B, Infected predominantly or exclusively with pre-C mutants, were given alFN 6-10 MU 3/w for 612 months. All pts were followed for at least one year after IFN withdrawal (range 1-6 years) with pedodic tests for serum ALT levels and HBV markers. Serum HBV-DNA was detected by spot hybridization, using a biotin-labeled 3.2-kb HBV genome. HBV-DNA pre-C sequence was amplified by PCR and directly sequenced by the dideoxynucleotide chain termination method. Results. At the end of therapy, 32/42 pts became HBV-DNA- and normalized ALT levels and were defined as responders. During follow-up, HBV reactivation and increased ALT levels occurred in 25•32 responder pts (78%), from 1 to 48 months after discontinuing IFN. In three pts HBV-DNA became undetectable after reactivation and ALT normalized for the rest of the follow-up period. Discussioq. These results demonstrate a high rate of initial response to alFN in anti-HBe positive pts, suggesting that IFN has an anti-viral effect on pre-C mutant viruses as on wildtype HBV. However, long term follow-up showed that a sustained response was present in only few pts, as evident from the frequent occurrence of reactivations after IFN withdrawal. Conclusions. Our data suggest that alFN therapy is less frequently able to induce a permanent remission in lots infected with pre-C mutants.
I
S Magrin, A Craxt. C Fabiano, L Marino, G Fiorentino, V Di Marco, O Lo lacono, P Almasio, MS Urdea*, JH Wilber*, C Bonura"*, F Gianguzza*", L Stuyver', L Pagliaro. Clinics Medica and ** Dip. Biol. Cell., Univ. Palermo; " Chiron Co.., USA; "Iunogenetics N.V., Belgium. Responsiveness to IFN in chronic hepatitis C differs according to HCV replication and genotype. We studied 99 anti-HCV +re Sicilian subjects (69 chronic hepatitis, CH; 30 cirrhosis, C). All received alpha2b IFN (10 MU tiw for 2 months,then 5 MU tiw for 4 to I0 months). ALT normalization on IFN was a primary response (PR), normal ALT after 12 months follow-up a sustained response (SR), any other pattern (NR). Serum HCV-RNA was found in 98/99 (99%) patients by nested PCR (5'UTR primers, sensitivity treshold 10-100 genome Eq/ml), and in 78/99 (78%) by branched DNA assay. Mean viraemia (xl06 Eq/ml by bDNA) was 7.7 in CH and 4.0 in C. HCV-RNA levels were 7,5 among the 49 PR and 5,7 among the 50 NR cases. Seventeen of the 49 PR became SR. Mean viraemia was lower in them (3.4). Specifically, 8/17 SR (47%) were bDNA negative as compared to 6/32 (18%) PR who relapsed and 7/50 (14%) NR cases. HCV was typed by a reverse-hybridization line probe assay (LiPA). Specificity was confirmed in some patients by sequencing. Type I was found in 85,99 [86%] subjects (4 la: 1 PR, 1 NIL 2 SR; 66 lb: 19 PR, 35 NIL 12 SR; 15 unclassified: 7 PR, 8 NR); type 2a in 4/99 [4%] (1 PR, 1 NIL 2 SR); type 3 in 4/99 [4%] (2 PR, 2 NR); type 4 in 6/99 [6%] (2 PR, 3 NIL 1 SR). Mean viraemias by type were: la: 3.8; lb: 6.7; 1 unclass.: 10.1; 2a: 0.4; 3: 2.22; 4: 5.5. CONCLUSIONS. Low viraemia correlates to SR. Genotype 1, mostly lb, is prevalent in Sicily. SR occurs more often with type la or 2a. Since most of our SR cases are infected by type lb, generally associated to unfavourable response in other areas, evaluation of other regions of the genome is needed to understand the relation between viral type and IFN response.
non-response
Posters 1
L, I P1 C1/05 I
[ P1 C1/07 I
IMMUNE MECHANISMSDURING INTERFERON.ALPHA (IFN) INDUCED VIRUSELIMINATIONIN CHRONICHBV AND HCV INFECTION:ANALYSIS OF VIRUS-SPECIFICCD4+ T LYMPHOCY'rES
HCV GENOTYPES IN PATIENTS WITH CHRONIC HEPATITIS C AND SILENT HCV INFECTION AFTER SUSTAINED BIOCHEMICAL RESPONSE TO INTERFERON
HM Diepolder, MC Jung, RM Hoffinao., R Zaehoval, FM Zwiebel, C Korherr, G Paumgarmer, G Riethmiiller, and GR Pape. luslitute of hnmunology and Dept. of Internal Medicine II, Klinikum GroBhadern. University of Munich, Germany. Virusspecific CD4+ T-I~mphoeytes are believed to be the mainstay of the amiviral immune response in several viral diseases, including influenza and acute hepatitis B. While [FN has been shown to be an effective treatment for a subgroup of patients with chronic hepatitis B (criB) and chronic hepatitis C (cHC). the role of virusspecific CIM+ T cells in viral elimination during IFN therapy is unknown. We serially studied the virusspecifie proliferative response of peripheral CD4+ T lymphocytes (CD4+T-R) to recombinant viral proteins in 10 patients with crib and 27 patients will1 cHC who completed a course of IFN therapy. None of tile patients with criB had a CD4+T-R to hepalitis B virus (HBV) core protein before IFN therapy. However. during IFN therapy those patients who developed a "flare" of liver inflammation and seroeonverted to anti-HBe transiently displayed a vigorous CD4+T-R Io HBV core protein. These results arc in agreement with studies that show an association of s CD4+T-R to HBe/core protein with viral elimination in acute hepatitis B as well" as spontaneous viral clearance in criB. In contrast, 13/27 patients with cHC showed a CD4+T-R to hepatitis C virus (HCV) core protein, NS3. NS4. or NS5 already before IFN therapy and sustained responders (SR) were confined to this group. A new CD4+T-R to HCV proteins during IFN therapy was typically present in transient responders during the intervall of response but was frequently lost again after relapse. Neither permanent or transient loss of HCV-RNA was preceded by an increased inflammatory activity. Our data suggest that a virus-specific CD4+T-R seems to be important for viral eliminiation during IFN therapy in both crib and cHC. However, while in criB an HBV-specific CD4+T-R is induced by IFN and accompanied by viral elimination, in eric only those patients with a high a priori CD4+T-R are apparently able to clear the infection. A detailed analysis of IFN induced immune mechanismsin cH'B and cHC may be a key for Ihe development of new rational therapies.
P.Pontisso t L.Chemello t C.Casarin t HG.Ruvoletto, H.Gerotto t A.Alberti. Clinica Hedica 2, University of Padova (Italy) Interferon treatment in patients with chronic hepatitis type C leads to complete response, in terms of biochemical remission, in about 25Z of the patients. ALT normalization is not invariably associated with viral clearance and serum HCV-RNA can persist after therapy in individual cases. To evaluate whether this behaviour is related to HCV genotype, 316 patients with chronic hepatitis C treated with interferon (3 to 9 HU/tiw for 6 to 12 months) and followed up for at least 12 months after therapy were studied. For genotyping, hybridization of the 5'UTR amplified products with probes specific for different HCV genotypes was used. Among 253 viremic patients, 46~ were infected by HCV-I (21Z la and 79Z Ib), 42Z by HCV-2, ii~ by HCV-3 and I~ by HCV-4. Long term response was observed in 85 patients, representing 9Z of HCV-I, 52~ of HCV-2 and 59Z of HCV-3 infected patients. Among these sustained responders with normal ALT, serum HCV-RNA persisted 6 to 12 months after therapy in 20Z of patients with HCV-I and in 45Z of those with HCV-2, while it was never found in those with HCV-3. Viral load was invariably decreased compared to basal levels. In conclusion, HCV-I and particularly HCV-2 but not HCV-3 are associated with clinically silent HCV infection after sustained biochemical response to interferon therapy.
I P1 c1/061
I Pl Cl/08
LONG-TERM RESPONSE TO INTERFERON THERAPY IN HEPATITIS B VIRUS (HBV) PRE-C MUTANTS ASSOCIATED CHRONIC HEPATITIS.
HCV VIRAEMIA, VIRAL GENOTYPES AND RESPONSE TO IFN IN PATIENTS WITH CHRONIC HEPATITIS C FROM SICILY.
T. Santantonio 1,2, M.Milella 1, L.Monno1' T.lacovazzi 1' G.Pastorel. H.Will 2 1Clinic of Infectious Diseases, Universita' dl Bad, Italy. 2Heinrich-Pette-lnstitut, University of Hamburg, Germany Objective. To evaluate the long-term therapeutic efficacy of alpha interferon (alFN) in chronic anti-HBe+ hepatitis B due to HBV pre-C mutants containing mutations in the pre-C region which prevent precora and e-antigen expression. Methods. 42 patients Cots) with anti-HBe/HBV-DNA+ chronic hepatitis B, Infected predominantly or exclusively with pre-C mutants, were given alFN 6-10 MU 3/w for 612 months. All pts were followed for at least one year after IFN withdrawal (range 1-6 years) with pedodic tests for serum ALT levels and HBV markers. Serum HBV-DNA was detected by spot hybridization, using a biotin-labeled 3.2-kb HBV genome. HBV-DNA pre-C sequence was amplified by PCR and directly sequenced by the dideoxynucleotide chain termination method. Results. At the end of therapy, 32/42 pts became HBV-DNA- and normalized ALT levels and were defined as responders. During follow-up, HBV reactivation and increased ALT levels occurred in 25•32 responder pts (78%), from 1 to 48 months after discontinuing IFN. In three pts HBV-DNA became undetectable after reactivation and ALT normalized for the rest of the follow-up period. Discussioq. These results demonstrate a high rate of initial response to alFN in anti-HBe positive pts, suggesting that IFN has an anti-viral effect on pre-C mutant viruses as on wildtype HBV. However, long term follow-up showed that a sustained response was present in only few pts, as evident from the frequent occurrence of reactivations after IFN withdrawal. Conclusions. Our data suggest that alFN therapy is less frequently able to induce a permanent remission in lots infected with pre-C mutants.
I
S Magrin, A Craxt. C Fabiano, L Marino, G Fiorentino, V Di Marco, O Lo lacono, P Almasio, MS Urdea*, JH Wilber*, C Bonura"*, F Gianguzza*", L Stuyver', L Pagliaro. Clinics Medica and ** Dip. Biol. Cell., Univ. Palermo; " Chiron Co.., USA; "Iunogenetics N.V., Belgium. Responsiveness to IFN in chronic hepatitis C differs according to HCV replication and genotype. We studied 99 anti-HCV +re Sicilian subjects (69 chronic hepatitis, CH; 30 cirrhosis, C). All received alpha2b IFN (10 MU tiw for 2 months,then 5 MU tiw for 4 to I0 months). ALT normalization on IFN was a primary response (PR), normal ALT after 12 months follow-up a sustained response (SR), any other pattern (NR). Serum HCV-RNA was found in 98/99 (99%) patients by nested PCR (5'UTR primers, sensitivity treshold 10-100 genome Eq/ml), and in 78/99 (78%) by branched DNA assay. Mean viraemia (xl06 Eq/ml by bDNA) was 7.7 in CH and 4.0 in C. HCV-RNA levels were 7,5 among the 49 PR and 5,7 among the 50 NR cases. Seventeen of the 49 PR became SR. Mean viraemia was lower in them (3.4). Specifically, 8/17 SR (47%) were bDNA negative as compared to 6/32 (18%) PR who relapsed and 7/50 (14%) NR cases. HCV was typed by a reverse-hybridization line probe assay (LiPA). Specificity was confirmed in some patients by sequencing. Type I was found in 85,99 [86%] subjects (4 la: 1 PR, 1 NIL 2 SR; 66 lb: 19 PR, 35 NIL 12 SR; 15 unclassified: 7 PR, 8 NR); type 2a in 4/99 [4%] (1 PR, 1 NIL 2 SR); type 3 in 4/99 [4%] (2 PR, 2 NR); type 4 in 6/99 [6%] (2 PR, 3 NIL 1 SR). Mean viraemias by type were: la: 3.8; lb: 6.7; 1 unclass.: 10.1; 2a: 0.4; 3: 2.22; 4: 5.5. CONCLUSIONS. Low viraemia correlates to SR. Genotype 1, mostly lb, is prevalent in Sicily. SR occurs more often with type la or 2a. Since most of our SR cases are infected by type lb, generally associated to unfavourable response in other areas, evaluation of other regions of the genome is needed to understand the relation between viral type and IFN response.
non-response