S232 POSTERS HCV infection. This critical role of the CD4+ T helper response is further supported by the association of certain HLA class II alleles and infection outcome. However, detailed information about many targeted CD4+ responses is limited and has restricted translational studies as well as the design of T-cell vaccines. We have combined an extensive HCV CD4+ T cell response database and patient HLA data with advanced computational methods to define novel HCV CD4+ epitopes out of an increasing number of newly identified CD4+ peptide responses. Methods: HCV specific CD4+ responses targeting the full HCV polypro- tein were identified using single peptides and short term Tcell lines from 100 subjects with different disease outcome (acute, resolved, HIV co- infected, chronic). Class II HLA typing was performed on all patients. HLA restriction predictions derived by computanional analysis were con- firmed by in vitro peptide HLA binding assays and cellular HLA restriction experiments. Class II restriction was further confirmed by generating class II tetramers for different HLA subtypes and successful staining of T-cell lines and PBMC. Results: Altogether 125 of 301 20-mer peptides were recognized by at least one subject. A majority of responses were located within the non- structural proteins and many were directed against a small number of highly conserved amd promiscuously binding CD4 epitopes. However, a large number of new epitopes were discovered and thus far we confirmed the HLA restriction of 8 of these epitopes. Interestingly, some, but not all tetramers showed cross-reactivity between different class II subtypes (e.g. DR0401 vs. DR0404). Discussion: The combination of a continuously expanding database of HCV CD4 epitopes that are targeted in vivo with cutting-edge computa- tional analysis of HLA associations and binding predictions accelerates the definition of HCV epitopes. This approach also allows a more complete appreciation of the promiscuity of CD4 epitopes. The results will allow new insights into the CD4 response that is critical for the outcome of HCV infection. 623 THE NEED FOR LIVER BIOPSY IS DECREASED USING A SIMPLE SCORE (FIBROFAST) OF HEPATIC FIBROSIS IN PATIENTS WITH CHRONIC HEPATITIS C G. Shiha 1 , W. Samir 2 , K. Zalata 3 , A. Elbeeh 4 , M. Zaki 4 , A. Attallah 4 . 1 Internal Medicine Department, Faculty of Medicine, 2 Biochemistry Department, Specialized Medical Hospital, 3 Pathology Department, Faculty of Medicine, Mansoura University, Mansoura, 4 Biotechnology Research Center, New Damitta, Egypt E-mail: g_shiha@hotmail.com Background and Aim: We published a simple non invasive score (Fibrofast) using five routine laboratory tests for the detection of significant hepatic fibrosis in patients with chronic hepatitis C (CHC) (Attallah et al., Hepatology Research (2006) 34: 163–169). The validation of 1067 cases from several international centers showed that the sensitivity of Fibrofast was 61.5%, specificity 81.1%, positive predictive value 59% and negative predictive value 82.6%. The performance of the test is not enough to suggestive as an alternative of liver biopsy. In this study we try to develop a new cut off score of the test that allows the diagnosis of established cirrhosis (F4) and (F0-F3) with the highest possible accuracy (more than 95%). Method: Subjects were 1266 patients with CHC. All biopsies were scored using METAVIR system. Our fibrosis score (Fibrofast) were measured and the performance of the new cut off score were done using ROC curve. Results: Liver biopsy showed that 1091 cases (F0-F3) and 175 cases established cirrhosis (F4). Using the ROC curve we develop new 2 cut off scores. The positive one is for diagnosis of (F0-F3) and the negative one is for diagnosis of established cirrhosis (F4). 112 out of 175 cases (64%) were established cirrhosis and 225 out of 1091 cases (20.6%) (FO-F3) i.e. 337 out 1266 cases (26.6%) was positively correlated with liver biopsy. r = 0.393, P = 0.0001, sensitivity 95%, specificity 95%. Conclusion: Fibrofast with the new two cut off scores could be an alternative to liver biopsy in about one third of the patients with sensitivity 95% and specificity 95%. Being non invasive, it could be an ideal marker for follow up during and after treatment in many patients. 624 HEPATITIS C VIRUS INFECTION ALTERS SERUM LEVELS OF THE MAJOR TYPE III INTERFERON IL-29 B. Langhans 1 , M. Michalk 1 , I. Braunschweiger 1 , G. Gornik 1 , N. Vidovic 2 , L. Leifeld 3 , J. Oldenburg 2 , T. Sauerbruch 1 , U. Spengler 1 . 1 Department of Internal I, 2 Institute for Experimental Hematology and Transfusion Medicine, University of Bonn, Bonn, 3 Evangelic Hospital K¨ oln Kalk, K¨ oln, Germany E-mail: spengler@uni-bonn.de;ulrich.spengler@ukb.uni-bonn.de Background and Aims: Interferons comprise several classes of cytokines and exert potent antiviral activity in viral hepatitis. Thus far, studies have been focused exclusively on type I and II interferons. Type III interferons, also designated as lambda-interferons, comprise cytokines IL-28A, IL-28B and IL-29, which are produced by plasmacytoid dendritic cells and bind to a shared receptor preferentially expressed on hepatocytes composed of IL10receptor2/IL28receptor alpha heterodimers. Methods: We studied serum levels of IL-29 and IL-28A in patients with acute replicative (n = 8), chronic persistent (n = 43) and spontaneously recovered self-limited (n = 24) hepatitis C by ELISA. Healthy uninfected subjects (n = 26) and patients with chronic hepatitis B (n = 16) served as controls. Results: IL-29 was detectable in all but 2 sera from healthy controls (167±57 pg/ml; Mean+/-SEM). Of note, IL-29 serum levels were signifi- cantly lower in patients with chronic hepatitis C (53±18 pg/ml; p = 0.035), while in acute hepatitis C IL-29 levels were intermediate between healthy controls and patients with chronic hepatitis C (123±84 pg/ml). Interest- ingly, IL-29 was normal in patients who had spontaneous viral clear- ance (208±69 pg/ml; p = 0.013 versus chronic hepatitis C) when com- pared to levels in healthy controls or patients with chronic hepatitis B (105±52 pg/ml). IL-29 tended to be lower in patients infected with genotypes 2 and 3 as compared to genotype 1 (25±22 pg/ml versus 36±11 pg/ml; p = 0.06) but did not show any correlation to serum viral loads. Serum levels of IL-28A were rather low as compared to IL-29 and did not reveal any significant differences between our study groups (healthy controls: 38±21 pg/ml, chronic hepatitis C: 24±11 pg/ml, acute hepatitis C: 33±10 pg/ml, self-limited hepatitis C: 79±34 pg/ml). Conclusion: Our data suggest that chronic hepatitis C is associated with reduced levels of IL-29. Intermediate IL-29 levels in patients with acute hepatitis C together with normal IL-29 levels after spontaneous recovery suggest that low levels of IL-29 mirror an impaired innate immune response against HCV. 625 POLYMORPHISMS IN TOLL-LIKE RECEPTORS 1 AND 2 ARE NOT ASSOCIATED WITH HEPATOCELLULAR CARCINOMA IN CHRONIC HEPATITIS C H.D. Nischalke, B. Reich, K. Aldenhoff, J. Nattermann, T. Sauerbruch, U. Spengler. University of Bonn, Department of Internal Medicine I, Bonn Germany E-mail: spengler@uni-bonn.de;ulrich.spengler@ukb.uni-bonn.de Background and Aims: Chronic hepatitis C virus (HCV) infection carries a high risk to progress to hepatocellular carcinoma (HCC). Recently, we and other groups have demonstrated that HCV proteins core and NS3 bind to TLR2 homo- and TLR2/TLR1 heterodimers and thereby induce several growth promoting cytokines. Polymorphisms in the TLR1 and TLR2 genes affect TLR expression and functionality and have been shown to predispose to various forms of cancer, such as gastric and colonic cancer or cancer of the prostate. Therefore, we studied if polymorphisms in the TLR1 and