April 1995 AASLD A l157

• RESPONSE 1N PATIENTS WITH HEPATITIS C VIRUS (HCV) INFECTION AND CRYOGLOBULINEMIA(CRYO) TO ALPH~ INTERFERON 2b (IFN). M Z Rofail, G Kerr, J H Lewis, D Kalloo, Z Buskell-Bales, L B Seeff Division of Gastroenterology and Rheumatology, VA Medical Center & Georgetown University, Washington, D,C.

Despite a high prevalence of HCV in mixed cryo patients, knowledge of the effect of therapy on the natural history of the disorder is limited. To help determine the efficacy of IFN on the clinical and biochemical parameters ofcryo in patients with chronic HCV, we retrospectively analyzed the results of treatment in a group with documented cryoglob- ulinemia. Methods We identified six patients undergoing IFN treatment for HCV- cryo.There were five males and one female, mean age 51 (42-67 years), five gave a history of IVDA and 4 consumed ETOH. Mean ALT was 112 IU/L (range 76-164) pretreatment. ANA was positive in one patient, 1:160 speckled pattern andrheumatoid factor was positive in another patient. All had symptoms including myalgias, arthralgias, Raynauds phenomenon, paraesthesias, and photosensitivity. Complete response to IFN was defined as normalization of ALT and loss of HCV-RNA for at least six months after the cessation of treatment. Symptom relief was a secondary endpoint. Results One of the six patients normalized ALT but remained HCV-RNA positive; two had a mild flare of ALT on IFN. Three of six had slight improvement of their rheumatologic symptoms, mainly myalgia and arthralgia. No patient had worsening ofcryoglobulin- related symptoms. Conclusion (1) Biochemical and serological complete response was not seen in this small group, but three of six had mild symptomatic improvement. (2) Long-term follow-up will be needed to define the natural history of cryoglobulin in HCV positive patients.

• BREAKTHROUGH IN PATIENTS WITH HCV CHRONIC HEPATITIS: ROLE OF GENOTYPES. L. Roffi, G Colloredo Mels*, G. Bdlati**, 13. Bmgnetti§, A. Ricci, A. Redaelli, A. Scalori, F. Panizzuti, M. Pozzi, A. Pipemo, P. Pioltelli, A. Bellobuono**, G Idco** and G. Mancia. Chair of Internal Medicine, S. Gemrdo Hospital, University of Milan; Monza (MI). *Dpt. Internal Medicine, Bolognini Hospital; Sedate (BG). **Dpt. Internal Medicine, Niguarda Hospital; Milan. §S. Pietro Hospital; Ponte S. Pietro (BG) Italy.

We have recently demonstrated that breakthrough (serum ALT increase more than two times over the upper normal limit during r-IFNa2 treatment atter an initial normalization) occurse in about 12% of patients with chronic hepatitis C (1). Development of neutralizing anti r-lFNa2 antibodies is associated with breakthrough in at least 50% of cases. In neutralizing antibodies negative patients other possible explanations include HCV escape mutants and r-IFNa induced down-regulation of specific IFN cell receptors. In chronic hepatitis C, distinct genotypes can be identified by sequence comparison of the viral Schemes and genotype may influence the clinical course of hepatitis and the response to IFN therapy.

Twenty-four patients experiencing breakthrough, and 31 responders and 28 non responders, consecutively enrolled in an open study, were studied. HCV genotyping was performed according to Okamoto, using RT-PCR with type- specific primers of the core gene (2). Neutralizing antibodies were measured accordin$ to a previously described method (1). Results arc shown in the table. GENOTYPE BREAK. RESPONDERS NON RESP. P I

I 3 3 0 as lI 8 9 16 ns Ili 8 9 9 ns IV 0 0 0 V 1 6 1

non typable 3 0 2 One patient with breakthrough and 4 responders had two distinct genotypes (III-V and II-III respectively). These differences are not significant.

In conclusion, breakthrough is a complex phenomenon in which different factors play a pathogenic role. Some of them (i.e. neutralizing antibodies to r-IFNa) have already been focused, while others need further investigations. According to our results genotypes seem to play no rote.

1 - L. Roffi et al.: Hepatology 21,3; 1995 (in press). 2 -H. Okamoto ct ah : J. Gen. Virol. 74 : 2385-90, 1993.

• T E C H N I C A L E V O L U T I O N S IN SPLIT LIVER TRANSPLANTATION

X. Rollers, M. Malag6, K.A. Gawad, J. Schulte am Esch, J. Erhardt*, M. Gt~tdlach and C.E. Broelsch Departments of Surgery, University Hospital Eppendorf, Hamburg and University Hospital, Essen*, Germany

From 01.01.94 to 13.12.94 s ixty-seven or thotopic liver transplantations were performed at the University of Hamburg. Of these, 14 (20.9%) were split liver transplantations (3 shipped from other centers) with a patient survival of 85,7 % (12/14) and a graft survival of 78,6 % (11/14). Eigth of these transplantations are the result of a new generation of techniques for splitting. Two livers were splitted using the in-situ splitting technique in the heart beating cadaveric donor. All 4 recipients are alive. One recipient of a left lateral segment suffered ischemic damage due to progressive portal steal after auxiliary grafting. After initial recuperation graft function is now progressively deteriorating. The others had an uncomplicated course and have normal liver function. Two livers were splitted ex-situ, using a technique derived from the in-situ technique, allowing to avoid all dissection of the bifurcation of the bile duct and to minimize manipulations to the hilar structures of the right lobe. All four patients had uncomplicated postoperative courses (including one with an auxiliary orthotopic graft and a right graft shipped to Essen). In summary, with the two new techniques, a 100% patient survival and a 100% graft survival (with probable functional loss of one graft) were obtained. Conclusions: 1. Split liver transplantation can be performed safely using the in-situ or modified ex-situ techniques. 2. ln-situ splitting allows shorter ischemic times and observation of the perfusion and hemostasis on both grafts before transplantation. 3. Shipping of splitted grafts between experienced centers is a valuable option.

VOLUME-ACTIVATED ION EFFLUX IN A HUMAN BILIARY CELL LINE. R. M. Roman, G. Shimokura, J. G. Fitz. Division of Gastroenterology, Duke University Medical Center, Durham NC.

Activation of ion transport mechanisms in biliary epithelial cells serves as a driving force for fluid and electrolyte transport. These cells are exposed to osmotic stress physiologically as a result of solute uptake and varying bile composition, and pathophysiologically due to ischemia and preservation/reperfusion injury. Our aim was to evaluate whether biliary cells possess ion effiux pathways that protect against cell swelling.

METHODS. Studies were performed in Mz-ChA-1 ceils derived from a human biliary adenocarcinoma which possess phenotypic features of biliary origin and regulated potassium (K +) and chloride (CI') channels. SrRb efflux and 1251 efflux were used as markers for changes in K+ and CI" permeability, respectively. Cell swelling was induced by exposure to hypotonic buffer (40 % decrease in NaCI).

RESULTS. Exposure of cells to hypotonic buffer caused a rapid increase in effiux of both SrRb and ~25I as compared to control cells in isotonic buffer. 1) SrRb efflux increased 53 % (p<.001) in hypotonic buffer; this increase was completely inhibited by the K ÷ channel blockers barium (3mM, p<.001) and quinine (lmM, p<.001). Ouabain (lmM) decreased basal but not volume-activated grRb effiux (p<.001). Bumetanide (250 uM) had no effect on SrRb effiux. 2) 12si effiux increased by 324 % (p<.001) in hypotonic buffer; this increase was inhibited by the CI" channel blocker NPPB (100uM, p<.01), but was unaffected by either barium or bumetanide.

CONCLUSIONS. Mz-ChA-1 cell swelling induced by exposure to a hypotonic environment stimulates effiux of both SrRb and ~2sI, consistent with activation of K ÷ and CI- channels. The increased rate of ion effiux is likely to contribute to volume regulation in biliary epithelia. Further characterization of the specific mechanisms involved may provide a focus for therapeutic intervention in such areas as biliary transport, and cholangiocyte ischemic and reperfusion injury.