HEPATOLOGY Vol. 22, No. 4, P t . 2, 1995 A A S L D A B S T R A C T S 461A

1 4 1 7 A N T I C A R D 1 O L I P I N A N T I B O D I E S A N D M I X E D CRYO- GLOBULINEM1A. L. Musset*, F. Lunel*, P. Cacoub*, P. Ghillani*, Z. Amoura**, J.P. Viard**, J.C. Piette*, P. Opolon*. *H6pital Piti& Salp~tri~re, **Hrpital Necker, Paris, FRANCE

Anticardiolipin antibodies (aCL) can appear in the course of various infectious diseases, but are not then associated with thrombotic events. Antibodies directed to B2 glycoprotein 1 (anti-82GPl), a co-factor of aCL, have been said to be associated with the presence of "autoimmune" aCE Recently, it has been demonstrated that about 50 % of "essential" mixed cryoglobuliuemia (MC) are associated with hepatitis C virus (HCV) infection. The aim of the study was to investigate aCL and anti-82GP1 antibodies in HCV patients with or without MC. Patients: We studied 3 groups of patients. Group 1:29 patients (18 M, 11 F) with chronic HCV infection and MC. Group 2 : 1 7 patients (10 M, 7 F) with chronic HCV infection hut without MC. Group 3:22 patients (12 M, 10 F) with essential MC and no HCV infection. In addition, 96 healthy blood donnors were used as a control group. Methods: Patients were considered to have MC if two successive determinations of their serum cryoglobulin level were above 0~05g/1. Anti-HCV antibodies were detected in all patients by third generation tests (ELISA, RIBA). aCL were detected using an ELISA (BMD Paris, France) and'the results were considered positive when above 14 GPL/MPL units (mean + 3 SD). Anti-B2GP1 were detected using an ELISA, and patients were considered positive if the OD was above 0.083 (mean + 3SD): In patients with MC, we also looked for aCL separately in Cryoprecipitate and in serum after extraction of MC, to investigate a possible "capture" of aCL in the cryoprecipitate. Results: lgG aCL were more frequently found in HCV patients than in control (9/46 (20%) vs 2/96 (2%); p<O.O01). However, the prevalence of aCL was similar in HCV patients with or without MC (6/29 vs 3/17; NS). None of the positive aCL patients had anti-112GPl. None of the positive aCL patients bad thrombotic events. In HCV patients (groups 1 and 2), the prevalence of aCL was not different whether patients were treated or not with interferon alpha. When we looked for the presence of aCL separately in serum and in cryoprecipitate, we did not find aCL in the cryoprecipitate. Conclusion: IgG aCL are frequently found in HCV patients (20%) whatever their status for MC. However, these aCL have the characteristics of infection-related aCL : low titer, absence of thrombotic events and absence of anti-g2GPl.

1 4 1 8 HEPATITIS-ASSOCIATED APLASTIC ANEMIA: V I R O L O G I C STUDIES AND LONG-TERM FOLLOW-UP. D Myerson, H-P Klein, CL Spurgeon, GB McDonald. Fred Hutchinson Cancer Research Center and University of Washington, Seattle, WA.

Hepatitis-associated aplastic anemia accounts for <5% of cases of severe aplastic anemia and is fatal in over 80% of patients unless treated. Marrow aplasia typically follows the onset of hepatitis by several months. Between 1971 and 1989, we treated 19 patients with this syndrome with allogeneic marrow transplantation. We studied pre- and post-transplant serum for evidence of infection by hepatitis viruses A, B and C and assessed long-term survivors for evidence of chronic liver disease.

METHODS. Patient charts were reviewed. Serum from 13 patients, collected before and after transplant and stored at -70 degrees C., was analyzed by nested polymerase chain reaction for HBV DNA and HCV RNA, respectively. Anti-HAV, anti-HBV and HBsAg were also determined pre-transplant.

RESULTS. At the time of initial diagnosis, patients were children or young adults (median 19 years, range 3 - 35) who presented with aplasia 0.5 to 8 months following clinical hepatitis (median 1 month). No patient had been transfused before the onset of hepatitis. The time from the onset of hepatitis to transplantation was 0.5 to 25 months (median 3 months). In 10 patients, hepatitis had resolved and liver tests were normal at the time of transplant. Transfusions had been given after development of aplasia but before transplantation in 16119 patients. Virologic studies before transplant were positive for anti-HAV (IgM) in 1/13 patients, for HBV DNA and HBsAg in 0113 patients, and for HCV RNA in 1/13 patients. At follow-up 4 - 23 years later, 15/19 are alive. Mild chronic hepatitis C and GVHD developed in 2/15 patients; the remainder currently have normal liver tests.

CONCLUSIONS. Hepatitis viruses A, B, or C are unlikely causes of hepatitis-associated aplastic anemia. There is a low frequency of chronic liver disease among long-term survivors, suggesting that the agent of acute hepatitis and marrow aplasia does not cause chronic infection.

1 4 1 9 COMBINATION THERAPY OF INTERFERON AND GLYCYRRHIZIN IN PATIENTS WITH CHRONIC HEPATITIS C. M. Naito. T. Kashiwaai. M. Naaasawa. T. T0yo, ma. Y. Tanaka. A. Ito. S. Ozaki. K~ Ishibashi.-M. Azuma. Department of Medicine, Osaka Kosei-Nenkin Hospital, Osaka, Japan.

Glycyrrhizin, a major component of a herb (licorice), has been widely used to treat chronic hepatitis B and C in Japan, because this substance has been shown to reduce aminotransferase levels. The purpose of this study was to investigate whether combination therapy of interferon (IFN)-a and glycyrrhizin could improve the beneficial effect of IFN alone therapy in chronic hepatitis C. METHODS Fifty-one chronic hepatitis C patients were included in this study (42 male and 9 female, mean age; 51.5 yr.). All patients received 9 million units (MU) of recombinant IFN-~-2a daily for 2 weeks followed 9 MU three times a week for 22 weeks (total dose; 720 MU). Twenty-six patients received 120 mg of glycyrrhizin on the same day of IFN administration, and the other 25 patients received IFN alone. There was no differences in the mean titer of hepatitis C virus (HCV) RNA between the both groups. The prevalence of subtype lb was greater in the combination therapy group (69%) than in the IFN alone group (52%). RESULTS At the end of therapy, 68% of patients achieved normalization of alanine aminotransferase (ALT) levels in the IFN alone group and 73% in the combination group. Among patients with subtype lb, the rate of ALT normalization was 54% in the IFN alone group and 67% in the combination group, furthermore, the rate of HCV RNA loss was 39% in the IFN alone group and 62% in the combination group. CONCLUSIONS The combination therapy of IFN and glychirrhizin might be efficacious for biochemical or virological response in patients with chronic hepatitis C.

1 4 2 0 DOES A PIGMENT STONE DIET PROMOTE NITROSAMINE- INDUCED HEPATOBILIARY MALIGNANCIES IN THE HAMSTER? A Nakeeb. Rtt Hruban*. PA Lin~tt. KD Lillemoe. HA Pitt. Department of Surgery and Pathology*, The Johns Hopkins Medical Institutions, Baltimore, Maryland.

Cholangiocareinoma is a rare tumor with an incidence of 1 per 100,000 population per year in the United States. However, biliary malignancies occur more frequently in Oriental countries where diets high in rice starch and low in protein may play a role in the pathogenesis of intrahepatic pigment stones and, possibly, cholangioearcinoma. Therefore, we tested the hypothesis that a pigment stone diet will promote the formation of tumors in a nitrosamine-induced hamster model of cholangiocarcinoma. Eight week old male Syrian golden hamsters were fed either a control chow diet (CHOW) or a high carbohydrate, low protein pigment gallstone inducing diet (PIG). In each group animals underwent weekly subcutaneous injection with either normal saline (NS) or the nitrosamine 2,2'-dihydroxy-di-n-propylnitrosamine (DIPN, 500 mg/kg) for ten weeks. At twenty weeks animals were evaluated for the presence of pigment gallstones and hepatobiliary malignancies. The results were:

Group N Stones Tumors

NS + CHOW 10 40% 0%

NS + PIG 10 70%* 0%

DIPN + CHOW 12 33% 17%

DIPN + PIG 12 67%* 8%

* p<0.05 vs CHOW, chi square Thes data suggest that a high rice starch, low protein diet 1 ) enhances pigment

gallstone formation but 2) does not increase the rate of DIPN-induced hepatobiliary tumors in the Syrian golden hamster. Therefore, we conclude that a pigment gallstone forming diet is not a promoter of nitrosamine-induced hepatobiliary malignancies.