HEPATOLOGY Vol. 22, No. 4, Pt. 2, 1995 AASLD ABSTRACTS 327A

8 8 1 RETREATMENT OF CHRONIC HEPATITIS B {CH HBV) WITH

ALPHA 2 INTERFERON (IFN) IN NONRESPONDERS AND RELAPSE PATIENTS IS EFFICIENT. JLPAYEN*. J IZOPgT**. K IMANI*r L ALRIC***.K BAPJkNGE*. M DUFFAUT***. JP VINEL*.

JP PASCAL*. J PUEL**. *Service d'H6pato-gaatroent6rologie et **Service de Virologie;cau Toulouse-Purpan 31059 Toulouse Cedex, ~**Service de M6decine Interne A,CHU Tculouse-Rangueil 31054 Toulouse Cedex-France.

We analyzed the retreatment (RT) of CH HBV with IFN to get the efficacy in nonresponders (NR) Or relapse patients (R). 95 patients with CH HBV were treated by IFN (5 MU tiw for 24 weeks) between November 90 and June 94. All patients were positive for HEs Ag, HBV-DNA and had histologically proved chronic active hepatitis. RT was achieved in 35 patients: 13 R patients (reappearence of HBVDNA after undetectable post-treatement period), 22 NR patients (persitent HBVDNA at the end of treatment) with IFN (Mean : 486 ± 222 mega units, for 25 ± 10 Weeks); the mean ALT level was 3.87 ± 2.5 N.We study: biochemical response (complete normalization of ALT), virological response:typeI:HBVDNA(-),without seroconversion; type II: HBVDNA(-I,with seroconversion e; type III: HBVDNA(-),with seroconversion e,s. Biochemical and virological relapse post RT. The mean follow up was 26.6±12 months.Results: in NR patients: biochemical response 12/22 (54%) followed by 3 (25%) relapse; virological response: type 1:2/22 (13.6%), type I~:4/22 (18.1%), type III:2/22 (9.1%), followed by 1 (11%) relapse. In the R patients: biochemical response 9/13 (69%) followed by 2 (22.2%) relapse; virological response: type 1:1/13 (7.7%), type II:7/13 (54%), type III: 0/13, followed by 3/8 (37.5%) relapse.Mean follow up post RT:10.2±8.Smonths. Conclusions: in nonresponders and relapse patients with chronic B hepatitis initialy treated with IFN alpha 2 (SMU tiw, 24 weeks), IFN retreatement with higher dose achieve a virological response in 41 % of nonresponders and 54 % of relapse patients.

882 TARGETED TREATMENT OF HEPATITIS B VIRUS (HBV) I N F E C T I O N BY A S I A L O - I N T E R F E R O N BETA AND ASSESSMENT O F ITS E F F E C T USING HBV NUDE MICE MODEL ESTABLISHED BY IN VIVO TRANSFECTION. T Eto. VS Trubetskoy*, VP Torchilin* and H Takahashi. Gastrointestinal Unit and *Department of Radiology, Harvard Medical School, Massachusetts General Hospital, Boston, MA

Hepatitis B virus (HBV) infection is a world wide health problem. In order to experimentally explore a new therapeutic regimens, we prepared asialo-interferon ~ (AS-IFN-15) to augment in vivo anti-viral effect of interferon treatment and tested its effect using an animal model of HBV infection established in nude mice by in vivo transfection. Methods: AS-IFN-~ was prepared from glycosylated natural IFN-13 (gift from M. Moriyama, TORAY Industries, Tokyo, Japan) by enzymatic treatment. This AS-IFN-~ demonstrated an antiviral effect similar to that of natural IFN-13 in vitro. Replication competent HBV plasmid DNA (head-tail homodimer of cloned HBV) was complexed with asialofetuin-poly-L- lysine conjugate and cationic-liposome to make virus-like particles for liver-specific transfection (infectious liposome). Asialoglycoprotein receptor-positive hepatoma cell were transfected with this infectious liposome complex more efficiently than cationic liposome alone or asialofetuin-poly-L-lysine alone. Infectious liposome complex was injected into the portal vein of nude mice via spleen. Seven days after in vivo transfection, mice were randomly selected and treated with intraperitoneal injections of physiological saline solution (PSS), IFN-I~ (1,000 or 10,000 IU/day) or AS-IFN-13 (1;000 or 10,000 IU/day) for seven days. In order to detect HBV virions in the serum from HBV- transfected mice, HBV particles in the test samples were first immunoprecipitated by anti-HBs Mab and HBV DNA in the immunoprecipitated particles was amplified by polymerase chain reaction. Results: HBV particles were produced in nude mice by in vivo transfeetion using infectious liposome. In nude mice treated with PSS, HBV viremia continued to the end of the treatment. In contrast, both IFN-I~ and AS-IFN-[3 demonstrated anti-viral effect and no HBV was detected by PCR in the mice sera after the treatment. Conclusions: Nude mice were successfully transfected by infectious liposome. Using this animal model, we demonstrated that AS-IFNq3 was effective to !nh;-bff I=I~V infecfio~ m

883 EXPRESSION OF THE INTERFERON-INDUCED ANTIVIRAL PROTEIN MXA IS INHIBITED IN VITRO BY THE HEPATITIS B VIRUS (HBV) CAPSID PROTEIN. O.Rosmorduc*-. H.Sirma L. P.Sonssan~ P.Lebon~ M.Horrisber~er$. C.Brechot Z. D.Kremsdorf*-. *INSERM U370, CHU Necker; Paris; §Laboratoire de Virologie, Hopital St Vincent de Paul, Universit4 Ren6 Descartes, Paris, France; $Ciba-Geigy; Basel; Switzerland.

We have previously shown that circulating defective hepatitis B virus particles (dHBV DNA), generated by singly spliced 2.2 kb HBV RNA, are associated with a chronic course of HBV infection. In addition, we have also shown that the in vitro expression of this dHBV DNA leads to overexpression of the HBV capsid proteins (HBcAg and HBeAg). We have examined whether these defective HBV particles might modulate antiviral effect of interferon alpha. Methods: Huh7 cells were stably transfected with dHBV DNA or the complete HBV DNA. We studied in these cells: I) global antiviral response to interferon a, by virus yield reduction experiments, using the vesicular stomatitis virus (VSV); 2) induction of 4 main proteins involved in the antiviral effect of interferon a: MxA protein, p69 and pl00 oligoadenylate- synthetases (OAS) and p68 protein kinase (PK), using western blot and semi- quantitative immunofhiorescence; 3) transcripts coding for the MxA protein, by northern blot. Results: 1) we observed more than 10 fold decrease in interferon a protective effect against VSV infection in dHBV DNA transfected ceils, as compared to HBV DNA transfected cells and negative controls. 2) we did not find a significant difference in the amount of induced PK or OAS. However, a significant reduction of MxA protein (about 3 fold), together with a decreased amount of the MxA RNAs, were observed in dHBV DNA transfected cells; 3) an inverse relationship between the iniracelhilar amount of HBV capsid protein and the IFN-induced MxA protein was also evidenced. Conclusions: Our results: 1) demonstrate that in vitro expression of dI-IBV DNA modulates the antiviral effects of the interferon alpha, through decreased MxA concentration; 2) suggest that the HBV capsid protein might be implicated in this inhibition, probably through transcriptional regulation. The mechanism of the MxA promoter activity modulation, as well as the protective effect of MxA on HBV genome expression, are now under investigation. Together, these findings might explain the absence of response to interferon in some HBV chronically infected patients.

884 HBV MARKER KINETICS DURING THERAPY OF CHRONIC HEPATITIS B: COMPARING INTERFERON (IFN) AND LAMIVUDINE (3TC). MC Kuhns. M Bartholomew*. M de Medina*. A McNarnara_ I Olson. P Green,and ER Schiff*. Abbott Laboratories, Abbott Park, IL and *Univ. of Miami School of Med. and VAMC, Miami, FL.

Virological response to antiviral therapy (Rx) for chronic hepatitis B is defined as sustained loss of HBeAg and serum HBV DNA (by direct hybridization). Anti-HBc IgM may also be usefulin monitoring response. We investigated the effects of 2 different therapies onthe kinetics of HBV DNA, HBeAg, and anti-HBc IgM. Study groups: 7 patients (4 responders,3 nonmsI~nders)treated for 16 weeks with IFN with or without prednisone withdrawal and 10 patients treated with 3TC for 12 weeks (2 at 25 rag,4 a t 100 mg, 4 at 300 rag). HBV DNA is rapidly lost during 3TC Rx in most patients (pts) but usually returns post-Rx; HBeAg loss occursin only 9% of pts (Hepatology 1994. 20(4):199A). Methods: HBV DNA quantitation by solution hybridization assay (Abbott), HBeAg quantitation by a new, prototype automated EIA with recombinant calibrators (AXSYM, Abbott); anti-HBc IgM index values determined by IMx Core M assay (Abbott). Results for 3 of the patient groups are summarized in the figures.

,.oI n.v OnA m~L H--, The initial decline in HBV DNA ~ , was greater with 3TC (all 1(301300

- - .~. mg pts) than with IFN. Although oa| .... .~ ~m.~4t, , 9/i0 3TC ptS remained HBeAg(+),

g[ | - - taavu~m~ ~" HBeAglevels in fact deelined ~ +o.++~ ~[~ :""8 :[ * " . during Rxbut more slowly than in ~ o+ x IFN pts. 5/5 IFN pts tested had

i~ x k (range 1.1-3.2 fold). Only4/103TC ~ ' ' ~ 1 pts showed an increase in anti-HBc

i S +2 l~IgM (range 1.2-2 fold) ineludingthe 1 nx wsm< nx wrmt( pt who lost HBV DNA and HBeAg.

Conclusions:A new prototype quantitative HBeAg assay (AXSYM) is a useful and practical method for monitoring the virological effects of Rx, especially in the evaluation of new regimens. Although HBV DNA toss OCCtLrt~ more rapidly with 3TC than with IFN , HBeAg declined more slowly with 3TC. Anti-HBc IgM increased during Rx in all IFN patients tested but in only 40% of 3TC patients.