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Introduction Changes in HBsAg plasma levels during antiviral therapy with pegylated interferon (IFN) predict response in adults with chronic hepatitis B (CH-B). Whether pre-treatment HBsAg plasma levels correlate with liver relaxed circular (RC) HBV DNA and covalently closed circular (ccc) DNA is controversial. To date, no information is available on HBsAg plasma level behaviour and response to treatment prediction in tolerant patients with infancy-acquired CH-B treated with lamivudine (LAM) and IFN. Aim To investigate whether HBsAg plasma levels predict response to LAM + IFN treatment in tolerant children with infancy-acquired CH-B, and to determine their association with plasma HBV DNA levels during treatment and with pre-treatment liver RC HBV DNA and cccDNA. Method Patients: 23 children (8 males, median age 10.2 yrs) with infancy-acquired CH-B (all HBeAg positive), treated for 52 weeks [lead-in LAM (3 mg/kg/d) for 9 weeks; LAM plus IFN-α (5 MU/m2 TIW) from week 9 for 44 weeks], were divided according to treatment response: 5 responders (R = anti-HBs seroconversion) and 18 non-responders (NR). Methods: Plasma HBsAg and HBV DNA levels were measured before (treatment week 0, TW0), during (TW9, TW28, TW52) and after (follow-up week, FUW24) therapy by Abbott ARCHTECT_ assay and real-time TaqMan PCR [both log10 IU/ml]. Baseline liver RC HBV DNA and cccDNA was quantified by real-time TaqMan PCR [copies/ng genomic DNA]. Results are presented as median. Results Baseline HBsAg levels were lower in R than NR (4.36 vs 4.74, p=0.02), but similar in R and NR at the end of LAM lead-in therapy (TW9) (4.34 vs 4.66, p=0.1). During IFN add-on therapy, at TW28 (2.34 vs 4.33) and TW52 (0 vs 4.08) levels were markedly lower in R than NR, the difference persisting at FUW24 (0 vs 4.51) (p

Introduction Plasma HBsAg reflects the transcriptional activity of covalently closed circular (ccc) DNA within the liver rather than the absolute amount of cccDNA copies. Higher plasma HBsAg levels are reported among tolerant than immuno-active HBeAg positive chronic hepatitis B (CH-B) patients. Only limited information is available on the interaction of plasma HBsAg, expression of HBsAg and HBcAg within the liver and liver relaxed circular (RC) HBV DNA and cccDNA in patients with infancy-acquired CH-B. Aims To evaluate whether there is a relationship between plasma HBsAg and HBV DNA levels and HBsAg/HBcAg expression within the liver and liver RC HBV DNA and cccDNA in tolerant children with infancy-acquired CH-B. Patients 23 children (eight males, median age 10.2 yrs) with infancy-acquired CH-B (all HBeAg+) in tolerant stage underwent liver biopsy prior to antiviral therapy with lamivudine and interferon-α. Methods Plasma HBsAg and HBV DNA levels were measured at therapy baseline by Abbott ARCHITECT® assay and real-time TaqMan PCR [both log 10 IU/ml]. Baseline liver RC HBV DNA and cccDNA was quantified by real-time TaqMan PCR [copies/ng genomic DNA]. Immunostaining of formalin-fixed, paraffin-embedded liver specimens assessed HBsAg and HBcAg expression [# of positive cells per 1000 hepatocytes]. Results are presented as median, range. Results Baseline plasma HBsAg levels were 4.67 (3.7–5.1), HBV DNA was 8.2 (7.1–8.9) both log 10 IU/ml and ratio HBsAg/HBV DNA 0.57 (0.44–0.65). Liver RC HBV DNA was 4.41 (2–5.3) log 10 copies/ng genomic DNA and cccDNA was 363 (0.3–3632) copies/ng genomic DNA. HBsAg is predominantly expressed in cytoplasm focally associated with membranous staining, 2.9 (0–56.9) positive cells/1000 hepatocytes, in contrast to HBcAg expressing predominantly in the nucleus, 8.3 (0.5–83.6) positive cells/1000 hepatocytes. There were positive bivariate Spearman correlations between plasma HBsAg levels and liver cccDNA (r=0.41, p=0.05), HBcAg liver expression and cccDNA (r=0.44, p=0.05), liver RC HBV DNA and cccDNA (r=0.67, p=0.04) and a trend towards correlation between plasma HBsAg and HBsAg liver expression (r=0.38, p=0.07). Conclusion Plasma HBsAg levels correlate with liver cccDNA in tolerant children with infancy-acquired chronic hepatitis B infection suggesting that plasma HBsAg levels acts as a surrogate marker of cccDNA within the liver in tolerant patients. Competing interests I Carey grant/research support from: BMS, Gilead, Y Zen: None declared, M Bruce: None declared, M Horner: None declared, S Bansal: None declared, D Vergani: None declared, G Mieli-Vergani: None declared.

Introduction Changes in HBsAg plasma levels during antiviral therapy with pegylated interferon (IFN) predict response in chronic hepatitis B (CH-B). High baseline plasma levels of inducible interferon-γ protein 10 (IP-10) predicted HBsAg loss in CH-B adults patients on nucles(t)ide analogues (NA) therapy. No data are available on the relationship between IP-10 and HBsAg plasma levels in children with infancy-acquired CH-B on combination therapy. Aims To investigate whether baseline plasma IP-10 levels and their changes during therapy can help to predict HBsAg loss in children with infancy-acquired CH-B on combination therapy with lamivudine (LAM) and IFN-α. Methods Patients: 23 children (eight males, median age 10.2 yrs) with infancy-acquired CH-B (all HBeAg+), treated for 52 weeks [lead-in LAM (3 mg/kg/d) for 9 weeks; LAM plus IFN-α (5 MU/m 2 TIW) from week 9 for 44 weeks], were divided according to treatment response: five responders (R = anti-HBs seroconversion) and 18 non-responders (NR). Methods Plasma IP-10, HBsAg and AST levels were measured in serial samples before (treatment week 0, TW0), during (TW9, TW28, TW52) and after (follow-up week, FUW24) therapy by ELISA [pg/ml], Abbott ARCHITECT® assay [log 10 IU/ml] and AutoAnalyser [IU/l]. The results are presented median. Results Baseline IP-10 levels were similar in R and NR (123 vs 99, p=0.4), but increased significantly in R than NR at TW28 (220 vs 79, p=0.04). Baseline HBsAg levels were lower in R than NR (4.36 vs 4.74, p=0.02), but similar in R and NR at TW9 (4.34 vs 4.66, p=0.1) and markedly lower in R than NR during IFN add-on therapy and therapy follow-up (TW28: 2.34 vs 4.33; TW52: 0 vs 4.08 and FUW24: 0 vs 4.51; p Conclusion Low baseline HBsAg levels, HBsAg plasma levels decline and IP-10 concentrations increase during interferon add-on at TW28 predict HBsAg loss and response to therapy in tolerant children with CH-B. Competing interests I Carey grant/research support from: BMS, Gilead, M Bruce: None declared, M Hussain: None declared, M Horner: None declared, S Bansal: None declared, D Vergani: None declared, G Mieli-Vergani: None declared.

Introduction Vertically acquired chronic hepatitis C (CH-C) is a mild disease in childhood but may accelerate in adolescence. Effective early therapy with pegylated interferon (Peg-IFN) and ribavirin (Riba) prevents progressive liver damage. Control of HCV infection depends on both innate and adaptive immunity. Little is known about treatment predictors in children with CH-C. Aim To assess whether single nucleotide polymorphisms (SNP) near IL28B gene rs12979860 and rs8099917, HCV genotype (G) and pre-treatment innate and adaptive immunity [frequency/phenotype of natural killer (NK) cells, HCV-specific T cell responses, frequency/phenotype of T regulatory cells (T-regs), plasma levels of IFN-γ inducible protein 10 (IP-10)] predict therapy response in children with CH-C. Method Patients: 32 children with CH-C (19 males, median age 12 yrs, 53% G1) treated according to genotype with Peg-IFN (60 μg/m 2 /week) and riba (15 mg/kg/d) were divided into responders (22, R), relapsers (4, Rel) and non-responders (6, NR). Methods: rs12979860 and rs8099917 were tested by direct sequencing; baseline numbers of NK cells (CD3 - CD56 + ) and their subsets CD56 dim /CD56 bright , CD3 − CD56 +/− CD16 +/− , of CD4+cells expressing programmed death receptor (CD4 + PD1 + ) and of T-regs (CD4 + CD25 + FoxP3 + ) by flowcytometry on peripheral blood mononuclear cells (PBMC). PBMC IFN-γ and IL-10 production after exposure to HCV-core, NS3, NS4, NS5 antigens was evaluated by intracellular cytokine staining. Baseline IP-10 plasma levels [pg/ml] were measured by ELISA. All presented as median. Results rs12979860 haplotype CC was present in 34% (91% R and 9% NR), CT in 47% (73% R, 7% Rel and 20% NR) and TT in 19% (17% R, 50% Rel and 33% NR); rs8099917 haplotype TT was seen in 50% (88% R and 12% NR), GT in 44% (50% R, 29% Rel and 21% NR) and GG in 6% (50% R and 50% NR) patients. Non-G1 CH-C was linked with better response than G1 (53% vs 80%, p=0.02). Baseline number of CD56 bright NK cells was higher in R than Rel & NR (3.7% vs 1.8% and 1.3%, p=0.05). Compared to R, Rel and NR had higher numbers of CD3 - CD56 - CD16 cells (17.4% vs 12.9% & 10.7%, p=0.05), of HCV-core-specific IL-10 producing cells (CD4 + /IL-10: 4.4% and 3.8% vs 1.7%, p=0.03), of CD4+PD1+ cells (7.1% and 6.9% vs 4.3%, p=0.03) and of T-regs (4.2% and 3.0% vs 1.4%, p=0.04). Baseline plasma IP-10 levels were higher in NR than Rel and R (85 vs 34 and 15, p Conclusion Possession of both major haplotypes CC rs12979860 and TT rs8099917 for IL28B gene SNPs and low baseline IP-10 levels predict successful therapy response in children with CH-C.