5 1 0 A A A S L D A B S T R A C T S HEPATOLOGY October 1995

1613 SIGNIFICANCE OF ANTIBODY TO ANTI ENVELOPE PEPTIDES IN CHRONIC HEPATITIS C VIRUS (HCV) INFECTION. V W ong , P Jackson, GIM Alexander . C a m b r i d g e Un i ve r s i t y School of Clinical Medic ine , Cambridge, CB2 2QQ

The relevance of antibodies to hepatitis C virus (HCV) envelope proteins is uncertain. To investigate this sera from 121 chronic HCV carriers (median age 37 years, range 22 to 77, including 33 females) were assayed for reactivity with 27 well conserved peptides ranging from 10 to 20 residues derived from E1 (amino acids 1-192) and E2/NS1 (amino acids 1-352) proteins of HCV. Peptides were synthesised and commercially cross linked to bovine se rum albumin through heterobifunctional cross-linker 3-maleimidobenzoic acid N- hydroxysuccimic ester or 2-iminothiolane hydrochloride. Antibody was sought us ing ELISA. Liver damage was scored according to stage of fibrosis 0-5; HCV RNA was sought by reverse transcription PCR us ing primers from the 5' un-translated region of HCV. Antibody to all except 2 conserved peptides was found in some patients with chronic HCV. The two commonest peptides inducing a serological response were a 16-mer from E1 (SGHRMAWDMMMNW- SPC) recognised by 23.3% of patients and a 18-mer from E2/NS1 (DLVNTNGSTHINRTALNC) recognised by 13.3%. However the presence of antibody to these peptides was not associated with biochemical markers of liver inflammation, liver fibrosis or the presence/absence of HCV RNA. In conclusion anti-bodies against conserved epitopes of E1 and E2/NS1 are identified in a minority of patients with chronic HCV infection but do not confer significant protection against viraemia or fibrosis.

1614 RAPID AND SUSTAINED D E C R E A S E IN BILIARY P H O S P H O L I P I D O U T P U T F O L L O W I N G 1 , 1 - DICHLOROETHYLENE (DCE). SH Woodard. L Kaphalia, A Wan_a. MT Moslen. University of Texas Medical Branch, Galveston, TX.

Background: DCE produces selective injury to the bile canalicular membrane of zone 3 hepatocytes within 2 hrs, Ultrastructural changes to the canaliculus include dilation of the canalicular lumen and blunting of the canalicular microvilli. DCE treatment also results in selective impairment of certain pathways of bile formation, notably decreases in organic anion, total biliary protein and pIgA outputs, while bile salt secretion is unchanged. We propose that canalicular injury results from the interaction of reactive DCE-glutathione (DCE-GS) conjugates with pericanalicular components; thus, DCE-induced injury could alter the function of the canalicular membrane mdr2 P-glycoprotein involved in phospbolipid transport. Objective: Our 0bjeetive was to test the hypothesis that DCE alters the biliary output of' phospholipids. Experimental Design: Freely moving male Sprague-Dawley rats equipped with a permanent biliary cannula for bile collection and a duodenal cannula for taurocholate infusion were treated with DCE (50 mg/kg po) or mineral oil vehicle. Results: Biliary output of phospholipids decreased to 25% basal by 30 minutes after DCE. Presence of DCE-GS conjugates within the canalicular lumen could inhibit mixed micelle formation therefore, we measured phospholipid output when biliary excretion of DCE metabolites was greatly diminished. Biliary phospholipid output remained at 50% basal levels for up to 10 hrs after DCE. Depletion of endogenous bile salts through bile collection Without tauroch01ate replacement caused a 5-fold decrease in biliary phospholipid output; DCE treatment of bile salt-depleted rats resulted in a complete absence of phospholipid in bile. Conclusions: Our observations of a sustained decrease in phospholipid output after DCE are consistent with an effect on the canalieular membrane mdr2 P- glycoprotein. (Supported by NIH DK 34806 and an Amaco Fellowship)

1615 LAMIVUDINE THERAPY OF FIBROSING CHOLESTATIC HEPATITIS (FCH) DUE TO RECURRENT HEPATITIS B VIRUS (I-IBV) POST-LIVER TRANSPLANTATION (LT). GM Woolf, FG Villamil, LM Petrovie, SE Rojter, L Sher, L Makowka, C Vieary, JM Vierling. Liver Transplantation Program Cedars-Sinai Medical Center/UCLA, Los Angeles, CA and Glaxo-WeUcome, Research Triangle Park, NC.

FCH is a fatal form of recurrent HBV infection post-LT that is unresponsive to medical therapy. Lamivudine competitively inhibits HBV DNA synthesis and replication. Preliminary data indicate that Lamivudine is a promising anti-viral agent for r~urrent I-IBV post-LT. We report the results of Lamivudine therapy in a 42 y.o Asian man with FCH who underwent LT in 1992 with HBIg prophylaxis for subfulminant hepatic failure due to HBV infection. Reinfection occurred 6 mos after discontinuation of I-IBlg 1 yr post-LT. Alfa-2b IFN (3MU tiw) treatment resulted in only a partial response. Liver biopsy showed chronic hepatitis. Liver tests alad HBV DNA levels rose, and the patient became severely fatigued. Repeat liver biopsy showed hepatocyte ballooning degeneration, bile ductular proliferation, focal bile plugs and early nodular regeneration. Immunostaining was positive for HBsAg and HBeAg. Lamivudine (100

was started and subset ~ results are summarized below. Lamivudine T Bili AST/)kLT Alk P y-gt Albumin HBV DNA

Therapy (m~/dl) (U/L) (U/L) U/L) (mg/dl) (pg/ml) Pre 1.8 147/183 234 1358 2.7 179

3 wks post 1.1 72/59 164 631 3.0 4.6 5 wks post 0.8 133/115 160 487 2.8 3.9 9 wks post 0.7 79/80 104 358 3.4 < 1.6 13 wks post 0.4 50/55 101 241 3.6 < 1.6 Liver biopsy after6 weeks of therapy showed cirrhosis with marked decrease in ballooning degeneration and inflammation. By week 13, liver tests were only mildly abnormal. HBV DNA became undeteetable by wk 7, although HBsAg and I-IBeAg remained positive. The patient's fatigue resolved and he has experienced no significant side effects: We conclude that Lamivudine therapy, in this patient, improved clinical symptoms, inhibited HBV DNA replication, significantly reduced liver test abnormalities and reversed the histopathologieal features of FCH.

1616 QUANTITATIVE LIVER FUNCTION TESTING PRE- AND POST- LIVER TRANSPLANTATION (LT). GM Woolf. DA Warmer. L Makowka, JM Vierling. Hepatology and Liver Transplantation Programs. Cedars-Siani Medical Center/UCLA, LA, CA, and Metabolic Solutions Inc., Merrimack, NH.

Currently, hepatic function post-LT is indirectly assessed by standard serum liver tests and volume of biliary drainage. In contrast, quantitative liver tests (QLT) directly assess metabolic functions and have been shown to correlate with disease activity. To study the utility of QLT pre- and post-LT, we evaluated liver function with the established monoethylglyeinexylidide (MEGX) test and a new breath test which measures hepatic oxidation of ~3C'phenylalanine (Phe). The MEGX test and the Phe breath test (PBT) assess microsomal and cytosolic enzyme functions, respectively. We studied 15 patients, 8 wks (range 1-32 wks) pre-LT and followed QLT post-LT for 8 wks. Liver diseases were classified as hepatocellular in 10 patients (HCV=7, alcohol=2, other=l) and cholestatie in 5 (PBC=3, PSC=2). Fourteen patients were cirrhotic (Childs-Pugh A=2, B=6, and C=6), and one had fulminant failure of indeterminate etiology. Post-LT courses were either complicated (moderate rejection, n=l or recurrent HCV, n=2) or unenmplieated (n=12, discharged from hospital within 14 days). QLT were performed while fasting and at rest. Serum MEGX levels, drawn 15 and 30 min after lidoeaine infusion (lmg/kg), were measured with a TDx Analyzer (Abbott Laboratories). Concurrently patients ingested |00 mg of 13C-pha and breath samples were obtained every 10 rain for lh for 3CO2 analysis using gas isotope rutio/mass spectroscopy'. Mierosomal and eytosolic functions were abnormal pre-LT as measured by the MEGX (14+16 ng/ml, nl > 70 ng/ml) and PBT (2.6+1.2%, nl > 4.0%) tests, respectively. MEGX and PBT values correlated pre-LT, at 1 wk and at 4-8 wks (r 2 =0.78~ p<0.05). MEGX (54+-35 ng/ml, but not PBT (2.9+1.8%) values, showed a significant difference between the pre- and post-LT tests in all patients (p<0.03). By 8 weeks p0st-LT, MEGX tests were near normal, while PBT had not improved. Complications of rejection and recurrent HCV infection were associated with. worsening of both QLT and improved with treatment. We conclude that: 1) the MEGX test and PBT correlate pre-LT and worsen in parallel during complications post-LT and 2) mierosomal function normalizes earlier than cytosolic function during recovery post-LT.