HEPATOLOGY Vol. 22, No. 4, Pt . 2, 1995 A A S L D A B S T R A C T S 4 0 7 A

1201 LIVER BIOPSY IN HEMOPHILIC PATIENTS WITH CHRONIC HCV INFECTION. JR Daruich, R Perez Biant;o, E Mi~nQr0. G Picchi0, M Tezanos Pinto, JA Findor. Hospital de Clfnicas, University of Buenos Aires. Instituto de Investigaciones Hematoldgicas Mariano R Castex, Academia Nacional de Medicina, Buenos Aires, Argentina.

Background: HCV infection is very frequent in hemophiliacs, but studies about its consequences are very scarce. Objective: To establish the frequency the liver damage in hemophiliacs, due to chronic HCV infection and its severity, 20 patients (pts) with HCV infection of at least six months duration were studied. Material and Methods: In 19 males and 1 female with mean age of 29.9 years (±14.9) a laparoscopy and liver biopsy, previous correction of the coagulation disorders, were performed, All the pts were anti HCV + r e (EIA 2 and RIBA 2) and HCVRNA +ve by n-PCR. Aminotransferase level was not considered as an inclusion criteria. Patients with HBV or HIV coinfection or history of alcoholism were excluded. Histologic specimens were analyzed by two independent observers and evaluated by the Knodell score. Results: No complications related to the procedure were observed. All but one of the pts presented with persistently normal aminotransferase level. In 2 (10%) cases laparoscopy showed the existence of cirrhosis. This finding was also confirmed by histology. In spite of the lack of elevation of the aminotransferases only 2 (10%) liver biopsies were considered normal. In the remaining biopsy specimens including the 2 (10%) pts with cirrhosis the Knodeil score showed values for the inflammatory component ranging between 2 and 9 and the fibrosis index between 1 and 4. Histologic markers considered to be closely related to HCV infection such as lymphoid aggregates or follicles in the portal tract, with or without germinal center were observed in 9 pts (45%), ductular damage in 9 (45%) and mild hepatocellular steatosis in 5 (25%). One or more of these three features were present in 13 pts (65%). Conclusions: In spite of the normal aminotransferase level histologic liver damage as consequence of chronic HCV infection =s a frequent event in hemophiliacs. This situation may evolve into a severe liver damage such as cirrhosis. The long term results of IFN treatment in this group of patients is now under evaluation.

1202 QUANTITATIVE DETECTION OF HEPATITIS C VIRUS AND GENOTYPES IN HAEMOPHILIC PATIENTS DURING

INTERFERON THERAPY C. Defer. L Lepot. A. Parquet. M. Manie,£. Centre Rtgional de Transfusion Sanguine, Lille, FRANCE.

We have determinated the hepatitis C virus (HCV) genotypes and followed the HCV-RNA levels of 8 anti-HCV positive haemophilie patients under interferon (IFN) therapy treated at the C.R.T.S. of Lille. Methods : The eight haemophiliacs were given 3 million units oflFN three times a week for 1 year. Serum samples for measurement of alanine aminotransferase (ALT) values were collected at week 0, regularly during the trial and 2 or 3 months after the trial had finished. We analyzed HCV- RNA levels by Amplicor HCV Monitor (Roche) and the genotyping was processed using HCV InnoLIPA assay (Innogeneties). Results :

Hemophiliac Genotype LB lb FP lb PG lb SJ lb

PJC 3a BC 3a QA 3a HC 3a

IFN response Genomes/mL at week 0 No No No Yes Yes Yes Yes Yes

406 211 l 576 383 960 286

5 691 59 084 57 959 207 191 277 778

Five patients had a complete response (normalization of ALT levels and undetectable HCV-RNA) but patient PJC relapsed 3 months after the end of the therapy. The follow-up of the other four is going on. Conclusion : The efficacy of IFN in chronic hepatitis C is dependent on the HCV-RNA levels and HCV genotype. In our study, generally, genotype lb is associated to high HCV-RNA levels (non responders) and genotype 3a to low HCV-RNA levels (responders). These findings have to be confirmed by the mean of a study on a larger population.

1203 EFFECT OF ISCHEMIC PRECONDITIONING ON LIVER INJURY INDUCED BY HYPOTHERMIC PRESERVATION. A STUDY IN THE ISOLATED PERFUSED RAT LIVER. N Dehnil= O Chazouill~res 2, M Vaubourdolle3= C.Rey z, P P De St Maur 4 C Housset 2, R Poupon2= L Hannoun t. 1Laboratoire d'Anatomie et d'Organogtn~se, ZlNSERM U402 et Unit6 d'Htpatologie, 3Service de Biochimie A, 4Service d'Anatomopathologie, Htpital Saint-Antoine, 75012 Paris, France.

We have previously shown that ischemic preconditioning (short periods of ischemia with intermittent reperfasion) has a protective effect in normothermic ischemia/reperfusion (I/R) liver injury. Because of its potential interest in liver transplantation, we therefore studied whether such an effect occurs in hypothermic I/R. Methods: Livers from 12 hr-fasted rats were excised, stored in UW solution at 4"C for 48 hrs and then repeffused for 90 min in a recireulating system with blood cells free oxygenated buffer (control group, n = 12). In the preconditioning groups, livers were subjected, by clamping the hepatic pedicle, to 1 (PC1, n = 9) or 2 (PC2, n = 8) in vivo periods of 10 min ischemia (followed by 10 min reperfusion) prior to cold storage and reperfusion. Perfusate activities of AST, LDH, CK (an index of endothelial injury), bile flow, portal flow and vascular resistances were sequentially determined. Liver biopsy was taken after Trypan blue perfusion at the end of the experiments. Resul ts : There was no significant differences among groups in hemodynamic parameters, enzyme releases or histological damage (hepatoeyte necrosis and sinusoidal cell staining by Trypan blue). Only bile flow was significantly increased during reperfusion in PC1 vs controls (p < 0.03, Mann-Whitney test). Conclusions: Preconditioning did not alter liver injury in this model of hypothermie I/R. When compared with our previous findings, this lack of protective effect might be related to 1) distinct mechanisms between normothermic and hypothermic I/R injury or 2) too long duration of hypothermic ischemia (48 hrs) inducing too severe damage (over the therapeutic window) or waning of the preconditioning effect.

1204 RELATIONSHIP BETWEEN HEPATOCYTE PROLIFERATIVE ACTIVITY AND LIVER FUNCTION IN HUMAN LIVER CIRRHOSIS. M Delhave, H Louis, C. Degraef, O. Le Moine, J. Devi~re, B. Gulbis, D. Jacobovitz, M. Adler, P. Galand. Dept. of Gastroenterology & Lab. Cyt. Cancer. Exp., Erasme Hospital, University of Brussels, Brussels, Belgium.

High hepatocyte proliferative activity in liver cirrhosis was shown to be related to the risk of developing hepatocellular carcinoma and decreased proliferation indices to patient's outcome in alcoholic hepatitis. This work aimed at evaluating the hepatocyte proliferative activity in an unselected population of cirrhotic patients regarding the severity &the disease. Fourty six cirrhotics (21 alcoholic, 20 viral and 5 other) were prospectively analyzed by proliferating cell nuclear antigen (PCNA) immunostaining on methanol-fixed, paraffin-embedded liver biopsies. In those conditions, PCNA- labelling index (PCNA-LI) measures the number of cells in S-phase and assessed tissue proliferation. The median value of the PCNA-LI for all samples was 4.3% (range : 0-20.2%); it declined with worsening Child-Pugh score : 9.15%, 5.3% and 2.4% in Child Class A, B and C respectively (p < 0.0005). A significant association was also observed between PCNA index and liver function assessed by the aminopyrine breath test (ABT) (p < 0.05).

PCNA-LI <4.3% Child-Pugh score" ABT b (hi > 4.5%) A (n) B (n) C (n) median (%) (range) 3 7 13 0.54 (0.16-3.61)

PCNA-LI > 4.3% 10 12 1 1.96 (0.37-6.05) a : p <0.05;b : p <0.05

The probability of survival was significantly higher in patients with a high PCNA-LI (> 4.3%) as compared to those with a lower PCNA-LI (at a median follow-up of 4.3 months : 0.95 and 0.58 respectively; p = 0.02). In 6 patients undergoing transjugular intrahepatic portosystemic shunt (TIPS) the PCNA-LI decreased after the procedure in all cases (from 2-9.6% to 0- 1.6%) (p < 0.005). This early impairment of hepatocyte proliferative activity after TIPS might reflect the functional alterations induced by this treatment. In conclusion, hepatocyte proliferative activity assessed by the PCNA-LI increased in cirrhotic patients but decreased with worsening of the disease.