- Email: [email protected]
Changes in the distribution of HCV genotypes during the last decades, among French patients with chronic hepatitis C
April 1995
AASLD
• CHARACTERIZATION OF QUALITATIVE AND QUANTITATIVE LYMPHOCYTE DIFFERENCES IN RESPONDERS AND NONRESPONDERS TO a-INTERFERON THERAPY FOR HEPATITIS C. FD Manela, GR Fernandez-Botran, S Cornell, CH Tamburro Liver Research Center, Depts. of Medicine and Pathology, Univ of Louisville, Louisville, KY. Hepatitis C infection leads to chronic disease in the majority of exposed individuals. Interferon-a2b (IFN-c0 treatment results in sustained or complete response in only 15-20% of treated patients. Although the underlying causes for this partial effectiveness are not clear, it is possible that differences in the immune response among chronic hepatitis C patients may contribute to treatment failure. The purpose of this study was to compare the lymphocytic responses in responders vs. non-responders to IFN-a therapy. Two groups of 4 individuals each, who had chronic hepatitis C with anti-HCV antibodies, positive HCV-RNA and elevated transaminases, treated with IFNc~ (3MU TIW x 6 months) were studied. Peripheral blood mononuclear cells (PBMC) were obtained from these patients and cultured in the presence of phytohemagglutinin [PHA] and pokeweed mitogen [PWM]. Proliferative responses were measured by [3H]-thymidine incorporation and the culture supernatants were assayed for the level of cytokines (IL-lg, TNF~, IL-2, IFNT, IL-4 and IL-10) by ELISA. No significant differences were observed in the mitogen-induced proliferative responses of responders vs. non-responders (Table 1). However, differences were evident in the levels of secreted cytokines in response to a suboptimal dose of PHA (Table 2). No differences were discernible at c ,timal stimulatory doses of PHA or PWM. TABLE 1
Control (cpm)
PHA 1 (cpm)
Non-responder
1166 + 859
40,139 _+ 36,088
Responder
2275 + 698
55,329 + 29,324
TABLE 2*
IL-1
,,Non-responder
22
13.3
54
22.5
Responder a
TNFc~
IL-10
IFN7
IL-2
IL-4
81.2
13.1
1
1
47.9 31.5 Baseline Levels
1
1
Significant stimulation of IL-1 and TNFa synthesis in the absence of IL2 and IL-4 up-regulation suggests that monoeytes may play an important role in effective response to IPNa therapy. Increased IFN3, production in responders suggests enhanced cytotoxic activity while increased IL-10 response in non-responders ma~/correlate with an immunosuppressive effect. These data suggest that monocyUc and cytotoxic T-cell responsiveness may characterize responders to IFN~ treatment.
• CHANGES IN THE DISTRIBUTION OF HCV GENOTYPES DURING THE LAST DECADES, AMONG FRENCH PATIENTS WITH CHRONIC HEPATITIS C. P. Marcellin, M. Martinot-Peignoux, M. Pouteau, C. Castelnau, N. Boyer, S. Erlinger. Service d'H~patologie et INSERM U24, Clichy, France. The distributin of HCV genotypes is influenced by geographical origin and possibly by the time of infection. The aim of this study was to determine the distribution of HCV genotypes in a population of patients with chronic hepatitis C and an history of blood transfusion or intravenous drug addiction, according to the estimated duration of HCV infection. Patients and methods. We studied 119 patients : 39 women and 80 men, mean age 42 years (19-72). Forty-four had an history of intravenous drug addiction and 75 had an history of blood transfusion. HCV genotyping was performed with reverse hybridization assay (LiPA) (INNO-LIPA, ImGeN). The classification of Simmonds et al. was used. The duration of HCV infection was estimated according to the date of transfusion or to the date of first illicit drug injection. Results. Distribution of HCV genotypes according to the duration of infection (years) 1b 3a others
<5 30% 40% 30%
5-15 34% 33% 33%
15-25 42% 25% 32%
> 25 53% 16% 31%
Conclusion. In French patients with chronic HCV infection acquired by intravenous drug addiction or transfusion, the proportion of genotype lb decreased and the proportion of genotype 3a increased during the last decades. These changes in the distribution of HCV genotypes might have consequences on the severity of the liver disease and the response to therapy.
Al117
DOES G E N O T Y P E A F F E C T THE COURSE OF HCV INFECTION ? A . M a n g i a , I . C a s c a v i l l a , G . L e z z i , R . C l e m e n t e , F.Spirito G.Maertens* , M.Rizzetto ° and A.Andriulli G a s t r o e n t e r o l o g y D i v i s i o n IRCCS "Casa Sollievo della sofferenza." S.Giovanni Rotondo, ITALY; *Innogenetics, G h e n t BELGIUM; °Gastroenterology Departement, M o l i n e t t e Hosp. Torino, ITALY D i f f e r e n t genotypes of H C V may have implications~ in the outcome of liver disease. According to, Simmonds' classification, g e n o t y p e ib has been m o r e frequently found in severe disease, w h e r e a s type 2~ seems to be associated with less p r o g r e s s i v e disease. AIM: to verify whether genotype's d i f f e r e n c e s affect the c l i n i c a l course of HCV infection. We s t u d i e d 70 H C V R N A +ve pts w i t h h i s t o l o g i c a l d i a g n o s i s of h e p a t o c e l l u l a r carcinoma (HCC) and 15 HCV R N A +ve a s y m p t o m a t i c pts (ALT every 2-4 m o . , m e a n follow-up 2 yrs) w i t h evidence of CPH or CAH in the liver. METHODS: sensitive n e s t e d PCR to amplify 5'UT region of HCV genome and reverse solid phase hybridization with type s p e c i f i c probes (Innolipa, Innogenetics). In some patients d i r e c t s e q u e n c i n g of 5"UT region has been done. RESULTS: among HCC pts g e n o t y p e ib has been found in 43% and type 2 in 40%; the r e m a i n i n g had m i x e d infection. A s y m p t o m a t i c c a r r i e r s show 82% of g e n o t y p e 2. .................................................. HCC asymptomatic ib(43%) 2a(40%) 2a(82%) m e a n age (yrs) 65 64 53 d u r a t i o n (yrs) 18 20 24 parenteral (%) 82 71 83 .................................................. CONCLUSION: Our results show: i) comparable p r e v a l e n c e of the two most c o m m o n g e n o t y p e in our region, in the HCC group; 2) g e n o t y p e 2 is largely p r e s e n t in a s y m p t o m a t i c group, but it does not p r e v e n t h e p a t i c damage; 3) a t r e n d to y o u n g e r age and longer d i s e a s e d u r a t i o n has b e e n found in a s y m p t o m a t i c pts w i t h type 2.
EFFECT OF H E P A T I C STIMULATOR SUBSTANCE A D M I N I S T R A T I O N ON C A D M I U M - I N D U C E D H E P A T O T O X I C I T Y IN THE RAT. A. Margeli, S. Theocharis, A.Constantinou, C. Spiliopoulou, M. Kokkala, A. Koutselinis. Dept. of Forensic M e d i c i n e and Toxicology, School of Medicine, U n i v e r s i t y of Athens, 75, M. Asias str., Athens, GR 115 27, Greece. The h e p a t o p r o t e c t i v e effect of hepatic stimulator substance (HSS) a d m i n i s t r a t i o n on c a d m i u m (Cd) induced h e p a t o t o x i c i t y was studied in the rat. The i n t r a p e r i t o n e a l a d m i n i s t r a t i o n of CdCI 2 at a dose of 2.5 m g / K g b o d y weight (b.w.) in male W i s t a r rats, caused acute liver injury e v a l u a t e d 24 hours after the administration. S e r u m enzyme activities of alanine aminotransferase, aspartate aminotransferase, alkaline p h o s p h a t a s e and histological p a r a m e t e r s were used for the e s t i m a t i o n of C d - i n d u c e d liver injury. HSS (LaBrecque et al, 1987, Hepatology, 7:100-106) at doses of i0, 20 and 40 m g / K g b.w. injected i0 hours after Cd administration, caused a d o s e - d e p e n d e n t d e c r e a s e of hepatic injury. Our results show that HSS a d m i n i s t r a t i o n i0 hours later than Cd injection, has b e n e f i c i a l effect against C d - i n d u c e d hepatotoxicity. Rats t r e a t e d w i t h the highest dose of HSS p r e s e n t e d reduced aspartate a m i n o t r a n s f e r a s e and alanine aminot r a n s f e r a s e levels, similar to those o b s e r v e d in Cdu n t r e a t e d rats. Cd a d m i n i s t r a t i o n caused chan~es in hepatocytes, including enlargement, n u c l e a r p y k n o s i s and focal necrosis a c c o m p a n i e d b y periportal and lobular inflammation. HSS administration caused r e c o v e r y of hepatic a r c h i t e c t u r e and d e c r e a s e of i n f l a m m a t o r y lesions in a d o s e - d e p e n d e n t manner. In conclusion, our results show that HSS p r o t e c t s the liver against C d - i n d u c e d injury. This r e s e a r c h was funded in part b y the ELPEN Pharmaceutical Company, Pikermi, Athens, Greece.
AASLD
• CHARACTERIZATION OF QUALITATIVE AND QUANTITATIVE LYMPHOCYTE DIFFERENCES IN RESPONDERS AND NONRESPONDERS TO a-INTERFERON THERAPY FOR HEPATITIS C. FD Manela, GR Fernandez-Botran, S Cornell, CH Tamburro Liver Research Center, Depts. of Medicine and Pathology, Univ of Louisville, Louisville, KY. Hepatitis C infection leads to chronic disease in the majority of exposed individuals. Interferon-a2b (IFN-c0 treatment results in sustained or complete response in only 15-20% of treated patients. Although the underlying causes for this partial effectiveness are not clear, it is possible that differences in the immune response among chronic hepatitis C patients may contribute to treatment failure. The purpose of this study was to compare the lymphocytic responses in responders vs. non-responders to IFN-a therapy. Two groups of 4 individuals each, who had chronic hepatitis C with anti-HCV antibodies, positive HCV-RNA and elevated transaminases, treated with IFNc~ (3MU TIW x 6 months) were studied. Peripheral blood mononuclear cells (PBMC) were obtained from these patients and cultured in the presence of phytohemagglutinin [PHA] and pokeweed mitogen [PWM]. Proliferative responses were measured by [3H]-thymidine incorporation and the culture supernatants were assayed for the level of cytokines (IL-lg, TNF~, IL-2, IFNT, IL-4 and IL-10) by ELISA. No significant differences were observed in the mitogen-induced proliferative responses of responders vs. non-responders (Table 1). However, differences were evident in the levels of secreted cytokines in response to a suboptimal dose of PHA (Table 2). No differences were discernible at c ,timal stimulatory doses of PHA or PWM. TABLE 1
Control (cpm)
PHA 1 (cpm)
Non-responder
1166 + 859
40,139 _+ 36,088
Responder
2275 + 698
55,329 + 29,324
TABLE 2*
IL-1
,,Non-responder
22
13.3
54
22.5
Responder a
TNFc~
IL-10
IFN7
IL-2
IL-4
81.2
13.1
1
1
47.9 31.5 Baseline Levels
1
1
Significant stimulation of IL-1 and TNFa synthesis in the absence of IL2 and IL-4 up-regulation suggests that monoeytes may play an important role in effective response to IPNa therapy. Increased IFN3, production in responders suggests enhanced cytotoxic activity while increased IL-10 response in non-responders ma~/correlate with an immunosuppressive effect. These data suggest that monocyUc and cytotoxic T-cell responsiveness may characterize responders to IFN~ treatment.
• CHANGES IN THE DISTRIBUTION OF HCV GENOTYPES DURING THE LAST DECADES, AMONG FRENCH PATIENTS WITH CHRONIC HEPATITIS C. P. Marcellin, M. Martinot-Peignoux, M. Pouteau, C. Castelnau, N. Boyer, S. Erlinger. Service d'H~patologie et INSERM U24, Clichy, France. The distributin of HCV genotypes is influenced by geographical origin and possibly by the time of infection. The aim of this study was to determine the distribution of HCV genotypes in a population of patients with chronic hepatitis C and an history of blood transfusion or intravenous drug addiction, according to the estimated duration of HCV infection. Patients and methods. We studied 119 patients : 39 women and 80 men, mean age 42 years (19-72). Forty-four had an history of intravenous drug addiction and 75 had an history of blood transfusion. HCV genotyping was performed with reverse hybridization assay (LiPA) (INNO-LIPA, ImGeN). The classification of Simmonds et al. was used. The duration of HCV infection was estimated according to the date of transfusion or to the date of first illicit drug injection. Results. Distribution of HCV genotypes according to the duration of infection (years) 1b 3a others
<5 30% 40% 30%
5-15 34% 33% 33%
15-25 42% 25% 32%
> 25 53% 16% 31%
Conclusion. In French patients with chronic HCV infection acquired by intravenous drug addiction or transfusion, the proportion of genotype lb decreased and the proportion of genotype 3a increased during the last decades. These changes in the distribution of HCV genotypes might have consequences on the severity of the liver disease and the response to therapy.
Al117
DOES G E N O T Y P E A F F E C T THE COURSE OF HCV INFECTION ? A . M a n g i a , I . C a s c a v i l l a , G . L e z z i , R . C l e m e n t e , F.Spirito G.Maertens* , M.Rizzetto ° and A.Andriulli G a s t r o e n t e r o l o g y D i v i s i o n IRCCS "Casa Sollievo della sofferenza." S.Giovanni Rotondo, ITALY; *Innogenetics, G h e n t BELGIUM; °Gastroenterology Departement, M o l i n e t t e Hosp. Torino, ITALY D i f f e r e n t genotypes of H C V may have implications~ in the outcome of liver disease. According to, Simmonds' classification, g e n o t y p e ib has been m o r e frequently found in severe disease, w h e r e a s type 2~ seems to be associated with less p r o g r e s s i v e disease. AIM: to verify whether genotype's d i f f e r e n c e s affect the c l i n i c a l course of HCV infection. We s t u d i e d 70 H C V R N A +ve pts w i t h h i s t o l o g i c a l d i a g n o s i s of h e p a t o c e l l u l a r carcinoma (HCC) and 15 HCV R N A +ve a s y m p t o m a t i c pts (ALT every 2-4 m o . , m e a n follow-up 2 yrs) w i t h evidence of CPH or CAH in the liver. METHODS: sensitive n e s t e d PCR to amplify 5'UT region of HCV genome and reverse solid phase hybridization with type s p e c i f i c probes (Innolipa, Innogenetics). In some patients d i r e c t s e q u e n c i n g of 5"UT region has been done. RESULTS: among HCC pts g e n o t y p e ib has been found in 43% and type 2 in 40%; the r e m a i n i n g had m i x e d infection. A s y m p t o m a t i c c a r r i e r s show 82% of g e n o t y p e 2. .................................................. HCC asymptomatic ib(43%) 2a(40%) 2a(82%) m e a n age (yrs) 65 64 53 d u r a t i o n (yrs) 18 20 24 parenteral (%) 82 71 83 .................................................. CONCLUSION: Our results show: i) comparable p r e v a l e n c e of the two most c o m m o n g e n o t y p e in our region, in the HCC group; 2) g e n o t y p e 2 is largely p r e s e n t in a s y m p t o m a t i c group, but it does not p r e v e n t h e p a t i c damage; 3) a t r e n d to y o u n g e r age and longer d i s e a s e d u r a t i o n has b e e n found in a s y m p t o m a t i c pts w i t h type 2.
EFFECT OF H E P A T I C STIMULATOR SUBSTANCE A D M I N I S T R A T I O N ON C A D M I U M - I N D U C E D H E P A T O T O X I C I T Y IN THE RAT. A. Margeli, S. Theocharis, A.Constantinou, C. Spiliopoulou, M. Kokkala, A. Koutselinis. Dept. of Forensic M e d i c i n e and Toxicology, School of Medicine, U n i v e r s i t y of Athens, 75, M. Asias str., Athens, GR 115 27, Greece. The h e p a t o p r o t e c t i v e effect of hepatic stimulator substance (HSS) a d m i n i s t r a t i o n on c a d m i u m (Cd) induced h e p a t o t o x i c i t y was studied in the rat. The i n t r a p e r i t o n e a l a d m i n i s t r a t i o n of CdCI 2 at a dose of 2.5 m g / K g b o d y weight (b.w.) in male W i s t a r rats, caused acute liver injury e v a l u a t e d 24 hours after the administration. S e r u m enzyme activities of alanine aminotransferase, aspartate aminotransferase, alkaline p h o s p h a t a s e and histological p a r a m e t e r s were used for the e s t i m a t i o n of C d - i n d u c e d liver injury. HSS (LaBrecque et al, 1987, Hepatology, 7:100-106) at doses of i0, 20 and 40 m g / K g b.w. injected i0 hours after Cd administration, caused a d o s e - d e p e n d e n t d e c r e a s e of hepatic injury. Our results show that HSS a d m i n i s t r a t i o n i0 hours later than Cd injection, has b e n e f i c i a l effect against C d - i n d u c e d hepatotoxicity. Rats t r e a t e d w i t h the highest dose of HSS p r e s e n t e d reduced aspartate a m i n o t r a n s f e r a s e and alanine aminot r a n s f e r a s e levels, similar to those o b s e r v e d in Cdu n t r e a t e d rats. Cd a d m i n i s t r a t i o n caused chan~es in hepatocytes, including enlargement, n u c l e a r p y k n o s i s and focal necrosis a c c o m p a n i e d b y periportal and lobular inflammation. HSS administration caused r e c o v e r y of hepatic a r c h i t e c t u r e and d e c r e a s e of i n f l a m m a t o r y lesions in a d o s e - d e p e n d e n t manner. In conclusion, our results show that HSS p r o t e c t s the liver against C d - i n d u c e d injury. This r e s e a r c h was funded in part b y the ELPEN Pharmaceutical Company, Pikermi, Athens, Greece.