Soluble CD30 serum levels in chronic hepatitis C before and after treatment with interferon-α

Soluble CD30 serum levels in chronic hepatitis C before and after treatment with interferon-α


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HEPATOLOGY October 2001



SOLUBLE CD30 SERUM LEVELS IN CHRONIC HEPATITIS C BEFORE AND AFTER TREATMENT W I T H INTERFERON-or. Enrico Di Cesare, Dipartimento Medicina Interna, Messina University, Messina Italy; Maria A Freni, Aldo Spadaro, Antonino Ajello, Nunziata Alessi, Maria L Resta, Oscar Ferrau', Dipartimento Clinico Sperimentale di Medicina e Farmacologia, Messina Italy

EFFECT OF TREATMENT OF CHRONIC HEPATITIS C ON T LYMPHOCYTES IN HIV-INFECTED HEMOPHILIACS: DECREASE IN ACTIVATED T CELLS IN SUSTAINED RESPONDERS. Silvia Sauleda, Centre de Transfusio i Banc de Teixits, Barcelona Spain; Isabel Caragol,Juan I Esteban, Alberto Juarez, Carmen Altisent, Isabel Ruiz, Teresa Espanol, Hospital Universitari Vall d'Hebron, Barcelona Spain; Lluis Pnig, Centre de Transfusio i Banc de Teixits, Barcelona Spain; Rafael Esteban, Jaume Guardia, Hospital Universitari Vall d'Hebron, Barcelona Spain

lnterferon-a (IFN-a) has been widely applied in the treatment of chronic hepatitis from virus C (HCV). Virus factors as well as the host immune response seem to influence the outcome of IFN-a therapy. ThlYrh2 balance is believed to play an important role in HCV infection and a Th2 secretory pattern has been shown to prevail in peripheral lymphocytes from chronically infected patients. CD30 is a glycoprotein preferentially expressed on membrane surface and released as soluble molecule (sCD30) by h u m a n T ceils producing Th2 type cytokines. In this study levels of sCD30 were measured in sera from 25 patients (M/F 17/8; median age 41 years, range 21-63) with HCV-related chronic active hepatitis (CAH) before and after a 6-month course of treatment with IFN-2a, 6 MU thrice weekly. Ten patients were sustained responders to the treatment (normal ALT and negative serum HCV RNA for at least 6 months after withdrawal of the drug) and 15 patients were non responders (high ALT levels and persistent HCV viremia during and at the end of the treatment). Twenty-one healthy blood donors (HBD), (M/F 16/5; median age 38 years, range 22-60), were used as control population. Before IFN-a treatment, patients with CAH had median sCD30 serum levels significantly higher (25.4 U/ML, range 4.8-245) than HBD (median 17.6 U/ML, range 1.5-41.9, p<0.04). No differences in age, gender, serum ALT values and histological activity index were found between CAH patients with sCD30 values above or within the upper limits of normal range (mean + 3SD in HBD). At the end of the IFN-R treatment, sCD30 values remained significantly higher than in controls (median 33.3 U/ML, range 5-186.6, p=0.0002). No differences were found in sCD30 levels both before and after IFN treatment between sustained responders and non responders. From this study, sCD30 serum values, although high in patients with HCV chronic hepatitis, do not seem to be correlated either with the activity of the disease or to the outcome of the IFN treatment.

Background: The outcome of antiviral treatment for chronic hepatitis C depends to a great extent on the patient's immune response. HCV coinfection has also been associated with lower immune restoration in HIV-infected patients receiving HAART. Aims: We have studied changes in T cell surface markers and intracellular cytokine expression in HIV positive hemophiliacs receiving interferon alfa-2b (IFN) plus ribavirin (RBV) for chronic hepatitis C. Subjects and methods: Twenty-two HtV HCV positive hemophiliacs (all 93CDC class A, 19 on HAART for 1.8 years) were treated with 3MU tiw iFN and 800 rag/day RBV (Intron A and Rebetol, Schering-Plough) for 6 to 12 months, of whom 9 were sustained responders (SR) and 13 non-responders (NR). We have investigated activation markers (DR, CD38) and intracellular cytokines expression (I1-2, TNFa, IFN?) in T cell subsets by flow cytometry at baseline, six months into IFN ÷ RBVtreatment and six months after end of combination treatment. Ten similarly treated hemophiliacs (5 SR and 5 NR) without HIV infection served as control. Results: At baseline the proportion of the different T cell subpopulations (CD4, CD8, CD4CD45RA, CD4CD45RO, CD8CD45RA, CD8CD45RO, CD8CD28-) was similar in all HIV infected patients and did not change during or after treatment. However, a significant decrease in the proportion of both CD4+DR+ and CD8+ CD38+ subsets, and a tendency towards a decrease in CD8+DR+, were observed in sustained responders at the end of follow up (See table). No such changes in activation markers were observed in HIV positive NR patients or in HIV negative control patients (CD4+DR+: 9%---8 at baseline vs 10%-+8 at end of follow up; CD8+CD38+: 55%+--14 vs 60+-5, irrespective of treatment outcome). Imracellular cytokine expression was not significantly modified by treatment. These changes in activation markers were unrelated to HIV replication level, CD4 cell count or antiretroviral regimen. Conclusions: Hepatitis C infection seems to contribute substantially to chronic immune stimulation in HIV-infected patients under HAARTand tb_issituation may be reversed after eradication of HCV following combination therapy. This might explain differences in the degree of immune restoration following HAART in coinfected patients. baseline 6 monthsIFN+RBV SR (n=9) NR O=13) SR NR CD4 540_+117 543±282 474_+t42 423±273 %CD4 27_+8 24_+11 31_+8 25_+12 %C04DR t3_+5 19_+10 1t+_3~ 20_+I0 %CD838 68_+12 69_+17 7I-+13 72-+14 %CDSOR 35+12 42+-19 2 7 _ + 1 3 38+-22 a: P=0.028;b: P=O.O11vs. baseline(Wileoxontest for p~iredsamples) oc P= 0.043; 13:P= 0.013 vs NR (MannWhitney U-test)

followup SR NR 712_+236 558-+250 30_+6 25_+10 7_+2~p 16-+9 54+1P 64_+26 2 3 _ + 1 1 38_+27



HISTOPATHOLOGY OF CHRONIC HEPATITIS C IN DIALYSIS PATIENTS. Heike A Erberich, Department of Pathology, University of Cologne, Cologne Germany; Hans - michael Steffen, Department of Internal Medicine, University of Cologne, Cologne Germany; Volker Dries, Hans Peter Dienes, Department of Pathology, University of Cologne, Cologne Germany

HCV EVOLUTION AND SERIAL CHANGE OF ITS TITERS IN BIOCHEMICAL RESPONDERS W I T H INTERFERON THERAPY. Shiro Murashima, Ryukichi Kumashiro . . . . . . . . . . Kurume University School of Medicine, Kurume Japan

Aims: Patients with renal failure and haemodialysis treatment are at high risk to develop chronic hepatitis C, and figures up to 40% have been reported in literature. Clinical data on this special group of patients have been published in several reports, Histological description and evaluation of liver biopsies proriding important clues to natural history of chronic hepatitis C however are infrequent and cover only few aspects. Methods: To this aim, we analysed liver biopsies of 20 patients with chronic hepatitis C and long-term haemodialysis due to renal failure compared to 20 non dialysis patients, matched in age, gender and duration of HCV-infection. All tissues were routinely prepared and different staging methods applied to facilitate a light microscopy diagnosis. In cases with doubtful virological status, H CV-testing by RT-P CR was performed. Analysis and evaluation of histopathology were performed according to the recommendations of Ishak et. al. especially regarding inflammatory activity. The stage of fibrosis was semiquantitativly scored using the proposal method of Chevallier et. al.. Statistic analysis (T-tests, correlation analysis, regression analysis) were preformed with SAS (Vers 8.0). The significant level was set at p-<.05 (/3=.85). Results: HCV-RNA was demonstrated in all sera and liver tissues by PCR amplification. All haemodialysis patients showed minimal to mild (grade 1-2) inflammatory activity, significant higher then in non dialysis patients (p = .01 ). The stage of fibrosis was the same in dialysis and non-dialysis patients. In relation to the inflammatory activity, the fibrosis stage was significant higher in dialysis then in non-dialysis patients (Index-score: ((HAIStage)2/(HAl-Stage)/2), p=.044). No correlation was found between inflammatory activity and stage of fibrosis, obviously indicating a lack of causal relationship. Step-wise regression analysis strengthens this observation by showing two different predictors: 1) for inflammatory activity the group-accompanying and 2) for stage of fibrosis the patients age. In addition we confirm that patients with renal failure and haemodialysis treatment are a high-risk group to acquire chronic hepatitis C. Conclusion: 1)The lower inflammatory activity in dialysis patients may be caused by the modified immunological response. 2)However, the comparable stage of fibrosis to that in non dialysis patients, seems to indicate a direct impact of hepatitis C infection on fibrogenesis.

AIM: Some patients show sustained biochemical responses without clearance of serum hepatitis'C virus RNA after interferon(IFN) therapy. However, its virological characteristics remain unknown. Thus, we evaluated serial changes in viral quasispecies and titers in such patients. Methods: We examined two typical cases of biochemical responders in which we could serially monitor arid take blood samples. We observed serial changes in quasispecies by PCR-SSCP in hypervariable region 1 (HVR-1). Furthermore, Virus titers were measured by competitive PCR (Amplicore Ver.2). Result: [Case.l, 54 y.o. Female (Fig.l)] Before IFN therapy, ALT was within the range of 20-40 IU with slight liver disorder. After 1FN administration, ALT rapidly normalized (10 IU). and the improvement of the liver function also corttinued for 12 months after the cessation of the therapy. Remarkable thing is that SSCP bands did not change and no quasispecies evofusions were observed durirlg the 12 months. Thereafter, with a relapse of liver function, 5SCP bands revealed that a major HCV variant, which had been present for abou t a year, disappeared and a denovo HCV variant appeared. As for viral tirers, during the 12 monthes of ALT normalization, virus titers were within the range of 110-320 KIU/mL, which is increased compare to those before the IFN therapy (30 KIU/mL). [Case. 2, 42 y.o. Male (Fig. 2) ] Before IFN therapy, ALT was within the range of 46-58 IU. During the IFN therapy, serum HCV disappeared immidiatsly, however ALT was still high. After the cessatio~ of the therapy, HCV RNA was soon detected in serum. In contrast, three months after the cessation of the therapy, liver function became normal for about a year. Also in this case, no changes in SSCP bands were observed during the period of ALT normalization. Virus titer during the period with normal liver function was higher (500-800 KIU/ml) than that before 1FN treatment (250 KIU/ml), . Discussion: Among biochemical re~ sponders, we found that HCV in the period of ALT normalization, show no viral evofusion. Futharmore HCV-RNA titer increased during the period. These results suggests that HCV escape host immune-surveillartce during the period. Fig. 1

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