Akinori Kasahara

Fas antigen expression in liver tissues of patients with chronic hepatitis B

BACKGROUNDnHepatitis B virus-infected cells can be eliminated by the cytotoxic T cell-mediated immune reaction. Fas ligand, recently detected on the surface of cytotoxic T cell, is thought to induce cells to apoptosis by adhering to Fas antigen.nnnAIMS/METHODSnTo evaluate the role of Fas antigen and apoptosis in chronic hepatitis B, we immunohistochemically studied Fas antigen and HBsAg expression in liver samples from patients with hepatitis B virus infection.nnnRESULTSnIn samples from 56 HBV patients, Fas antigen was mainly expressed in the cytoplasm (partly at the membrane) of hepatocytes, and these positive cells were detected especially at the periportal region near piecemeal necrosis. According to Knodells HAI scoring system, the scores of periportal inflammation and necrosis (category I) and the scores of intralobular inflammation and necrosis (category II) were similarly higher in Fas antigen-positive cases than in Fas antigen-negative cases (p < 0.01), and there was a positive correlation between these scores and the degree of Fas antigen expression. In normal cases, Fas antigen was not detected. In patients with HBV infection, Fas antigen expression was closely correlated with the activity of the viral hepatitis. HBsAg was expressed by the majority of hepatocytes. However, Fas antigen was expressed by fewer hepatocytes than the number of HBsAg-positive cells.nnnCONCLUSIONSnThese findings suggest that the expression of Fas antigen may not be triggered only by HBV infection, and immunological interaction may be needed for the expression and for apoptosis to occur.

Circulating matrix metalloproteinase-2 and tissue inhibitor of metalloproteinase-1 as serum markers of fibrosis in patients with chronic hepatitis C : relationship to interferon response

BACKGROUND/AIMS/METHODSnThe imbalance between matrix metalloproteinases (MMPs) and tissue inhibitors of matrix metalloproteinases (TIMPs) is considered to be an important determinant of extracellular matrix deposition and breakdown. We measured serum MMP-1, MMP-2, TIMP-1 and TIMP-2 levels using the respective one-step sandwich enzyme immunoassays in 98 patients with chronic hepatitis C treated with interferon beta to examine their clinical significance for assessment of liver histology and to determine whether they can be useful as predictors of the interferon response.nnnRESULTSnSerum TIMP-1 levels showed a positive correlation with the degree of fibrosis (r(s)=0.30, p= 0.004). Serum MMP-2 levels revealed positive relationships with the degree of periportal necrosis (r(s)= 0.32, p=0.002), the degree of fibrosis (r(s)=0.26, p= 0.01) and total score of histological activity index (r(s)=0.24, p=0.02). Serum MMP-2 levels were significantly higher in patients with no response than in those with sustained and transient response (p<0.01 and p<0.05, respectively), while serum MMP-1 levels did not differ among the three groups. Compared with the levels in sustained responders, the total amounts of serum TIMP-1 were significantly lower in transient responders and non-responders (p<0.01 and p<0.001, respectively). As for serum TIMP-2 levels, a significant decrease was found in transient responders and non-responders (p<0.01). The ratios of serum MMP-2 to TIMP-1 levels were significantly higher in transient responders and non-responders than in sustained responders (p<0.001, respectively) even when HCV RNA levels were low in patients with HCV genome subtype 1b or when the HCV genome subtype was 2a or 2b. Sustained response was never found in type 1b patients with ratios of serum MMP-2 to TIMP-1 levels of over 6.0. In logistic multivariate regression analysis, the ratios of serum MMP-2 to TIMP-1 level (p=0.0001), HCV genome subtype (p=0.005) and serum TIMP-2 level (p=0.03) were the independent predictors for sustained response, while serum MMP-2 level (p=0.0006) was the only predictor for no response.nnnCONCLUSIONSnSerum MMP-2 and TIMP-1 levels might be useful for estimating the degree of liver fibrosis. The ratio of serum MMP-2 to TIMP-1 levels may serve as a new predictor of interferon response in patients with chronic hepatitis C.

Site-specific analysis of N-glycans on haptoglobin in sera of patients with pancreatic cancer: a novel approach for the development of tumor markers.

It was found in our previous studies that the concentration of fucosylated haptoglobin had increased in the sera of patients with pancreatic cancer (PC) compared to those of other types of cancer and normal controls. Haptoglobin, an acute phase protein, has four potential N‐glycosylation sites, although it remains unknown which site is responsible for the change in fucosylated N‐glycans. In the present study, site‐specific N‐glycan structures of haptoglobin in sera obtained from patients with PC or chronic pancreatitis (CP) were analyzed using liquid chromatography‐electrospray ionization mass spectrometry. Mass spectrometry analyses demonstrated that concentrations of total fucosylated di‐, tri‐ and tetra‐branched glycans of haptoglobin increased in the sera of PC patients. Tri‐antennary N‐glycans containing a Lewis X‐type fucose markedly increased at the Asn211 site of haptoglobin N‐glycans. While fucosylated N‐glycans derived from serum haptoglobin of patients with CP slightly increased, di‐fucosylated tetra‐antennary N‐glycans were observed only at this site in PC patients, and were absent in the haptoglobin of normal controls and individuals with CP. Thus, the present study provides evidence that site‐specific analyses of N‐glycans may be useful as novel tumor markers for PC.

Effect of interferon therapy on the incidence of hepatocellular carcinoma and mortality of patients with chronic hepatitis C: a retrospective cohort study of 738 patients.

The effect of interferon on the long‐term clinical outcome of patients with chronic hepatitis C remains unclear. This study included 594 patients with chronic hepatitis C who received interferon‐α therapy (Interferon group) and 144 patients with chronic hepatitis C who did not receive interferon (Control group). The patients in the Interferon group were classified into the following three groups based on the response of the serum aminotransaminase level of the patient during and after completion of the therapy protocol: sustained responders (n = 175), transient responders (n = 165), and non‐responders (n = 254). The age, sex, serum aminotransaminase level, platelet count, histological staging, hepatitis C virus (HCV) subtype, and HCV concentration at baseline were adjusted with the Cox proportional hazards model. The length of follow‐up for assessment of the risk for developing hepatocellular carcinoma (HCC) was 57.2 ± 13.9 months in the Interferon group and 67.7 ± 28.7 months in the Control group. Multivariate analysis showed that interferon therapy decreased the risk for developing HCC by 48% compared with that in the Control group (P = 0.064). The older the age, being male, having a low platelet count, and higher histological stage were independent factors associated with the development of HCC. The hazard rate ratio for development of HCC in the sustained responders, transient responders, and non‐responders was 0.16 (95% confidence interval [CI]: 0.04–0.62), 0.27 (95% CI: 0.09–0.79), and 0.74 (95% CI: 0.37–1.48), respectively. During follow‐up, 18 patients in the Interferon group died (10 from liver‐related diseases) and 17 patients in the Control group died (10 from liver‐related diseases). No sustained responder or transient responder in the Interferon group died of liver‐related disease. The cumulative survival rates of the Interferon and Control groups were nearly identical during the first 5 years following diagnosis. Thereafter, the cumulative survival rate of the Control group declined, resulting in an 8‐year survival rate in the Interferon and Control groups of 97% and 81%, respectively (P = 0.061). Similar trends were seen in the survival analysis of those who had died of liver disease: the 8‐year survival rates of the Interferon and Control groups were 98% and 88%, respectively (P = 0.32). Our study demonstrated that interferon therapy significantly lowered the incidence of HCC among patients with chronic hepatitis C who showed sustained normalization and among patients who showed transient normalization of the serum aminotransferase level after completion of interferon therapy. The survival analyses and determination of cause of death suggested that interferon therapy improves the long‐term survival of chronic hepatitis C patients who respond to this therapy, possibly by decreasing mortality from liver‐related diseases. Int. J. Cancer 87:741–749, 2000.

Ribavirin dose reduction raises relapse rate dose-dependently in genotype 1 patients with hepatitis C responding to pegylated interferon alpha-2b plus ribavirin

Summary.u2002 The impact of ribavirin exposure on virologic relapse remains controversial in combination therapy with pegylated interferon (Peg‐IFN) and ribavirin for patients with chronic hepatitis C (CH‐C) genotype 1. The present study was conducted to investigate this. Nine hundred and eighty‐four patients with CH‐C genotype 1 were enrolled. The drug exposure of each medication was calculated by averaging the dose actually taken. For the 472 patients who were HCV RNA negative at week 24 and week 48, multivariate logistic regression analysis showed that the degree of fibrosis (Pu2003=u20030.002), the timing of HCV RNA negativiation (Pu2003<u20030.001) and the mean doses of ribavirin (Pu2003<u20030.001) were significantly associated with relapse, but those of Peg‐IFN were not. Stepwise reduction of the ribavirin dose was associated with a stepwise increase in relapse rate from 11% to 60%. For patients with complete early virologic response (c‐EVR) defined as HCV RNA negativity at week 12, only 4% relapse was found in patients given ≥12u2003mg/kg/day of ribavirin and ribavirin exposure affected the relapse even after treatment week 12, while Peg‐IFN could be reduced to 0.6u2003μg/kg/week after week 12 without the increase of relapse rate. Ribavirin showed dose‐dependent correlation with the relapse. Maintaining as high a ribavirin dose as possible (≥12u2003mg/kg/day) during the full treatment period can lead to suppression of the relapse in HCV genotype 1 patients responding to Peg‐IFN alpha‐2b plus ribavirin, especially in c‐EVR patients.

Interferon treatment improves survival in chronic hepatitis C patients showing biochemical as well as virological responses by preventing liver-related death

Summary.u2002 Interferon therapy for chronic hepatitis C reduces the risk of hepatocellular carcinoma, especially among virological and biochemical responders. However, little is known about the effect of interferon therapy on mortality. We studied the long‐term effect of interferon therapy on mortality in patients with chronic hepatitis C. For this retrospective cohort study, 2954 patients with chronic hepatitis C were recruited, of whom 2698 received interferon therapy and 256 did not. The effect of interferon therapy on survival was assessed by standardized mortality ratio (SMR) based on published mortality data for the general Japanese population and by risk ratio calculated by proportional hazard regression. Over 6.0u2003±u20032.2u2003years follow‐up, death from liver‐related diseases was observed in 69 (68%) of 101 deaths among interferon‐treated patients and in 42 (81%) of 52 deaths among untreated patients. Compared with the general population, overall mortality was high among untreated patients (SMR: 2.7; 95% CI: 2.0–3.6) but not among interferon‐treated patients (SMR: 0.9; 95% CI: 0.7–1.1). Liver‐related mortality was extremely high among untreated patients (SMR: 22.2; 95% CI: 16.0–30.0) and less among interferon‐treated patients (SMR: 5.5; 95% CI: 4.3–6.9). The risk of death from all causes was lower for interferon‐treated than untreated patients (risk ratio: 0.47; 95% CI: 0.261–0.836; Pu2003=u20030.01). The risk of death from liver‐related diseases was significantly lower for sustained virological responders (risk ratio: 0.04; 95% CI: 0.005–0.301; Pu2003=u20030.002) compared with untreated patients, but not for nonsustained virological responders. Sustained biochemical responders (risk ratio: 0.03; 95% CI: 0.004–0.230; Pu2003<u20030.001) and transient biochemical responders (risk ratio: 0.18; 95% CI: 0.063–0.532; Pu2003=u20030.002) showed a significantly reduced risk of death from liver‐related death, whereas biochemical nonresponders did not. Hence interferon treatment improved survival in chronic hepatitis C patients showing a biochemical as well as a virological response by preventing liver‐related deaths.

Serum levels of soluble Fas antigen in chronic hepatitis C patients

Background/Aims: In chronic hepatitis C, the expression of Fas antigen on hepatocytes is upregulated and Fas ligand expression is detected on liver-infiltrating mononuclear cells. Thus Fas antigen/Fas ligand-mediated apoptosis is thought to be involved in hepatic injury in chronic hepatitis C. The soluble form of Fas antigen has been detected in serum and shown to inhibit Fas-mediated apoptosis. The present study was done to evaluate the relationship of serum soluble Fas antigen levels with disease activity. Methods: Serum soluble Fas antigen levels were measured by enzyme-linked immunosorbent assay for 68 chronic hepatitis C patients and compared with those in normal volunteers, chronic hepatitis B patients and autoimmune hepatitis patients. These levels were compared with histological activity, ALT levels, HCV-RNA titer and Fas expression on hepatocytes. Results: Serum soluble Fas antigen levels in chronic hepatitis C patients (3.24 ± 1.55 ng/ml) were significantly higher than those in normal volunteers (1.70 ± 1.01 ng/ml) (p < 0.01). They showed no difference from those in chronic hepatitis B or autoimmune hepatitis patients. Histologically, soluble Fas antigen levels showed correlation with the levels of liver inflammation (p < 0.01). However, no relationship was observed between serum soluble Fas antigen and serum ALT levels or HCV-RNA titer. Serum soluble Fas antigen levels showed correlation with the levels of Fas antigen expression in liver tissue (p < 0.05). Conclusions: These findings suggest that serum soluble Fas antigen may reflect the expression levels of Fas antigen on hepatocytes and the severity of liver inflammation in chronic hepatitis C.

Long-term effect of lamivudine treatment on the incidence of hepatocellular carcinoma in patients with hepatitis B virus infection

BackgroundNucleotide analogues have recently been approved for the treatment of patients with hepatitis B virus (HBV) infection. However, it is still controversial whether the decrease of HBV-DNA amount induced by treatment with nucleotide analogues can reduce the risk of hepatocellular carcinoma (HCC) development in HBV patients.MethodsA total of 293 HBV patients without HCC who were treated with lamivudine (LAM) were enrolled in a multicenter trial. The incidence of HCC was examined after the start of LAM therapy, and the risk factors for liver carcinogenesis were analyzed. The mean follow-up period was 67.6xa0±xa027.4xa0months.ResultsOn multivariate analysis for HCC development in all patients, age ≥50xa0years, platelet count <14.0xa0×xa0104/mm3, cirrhosis, and median HBV-DNA levels of ≥4.0xa0log copies/ml during LAM treatment were significant risk factors. The cumulative carcinogenesis rate at 5xa0years was 3% in patients with chronic hepatitis and 30% in those with cirrhosis. For the chronic hepatitis patients, the log-rank test showed the significant risk factors related to HCC development to be age ≥50xa0years, platelet count <14.0xa0×xa0104/mm3, and hepatitis B e antigen negativity, but median HBV-DNA levels of <4.0xa0log copies/ml (maintained viral response, MVR) did not significantly suppress the development of HCC. In cirrhosis patients, however, the attainment of MVR during LAM treatment was revealed to reduce the risk of HCC development.ConclusionsThese results suggest that the incidence of HCC in HBV patients with cirrhosis can be reduced in those with an MVR induced by consecutive LAM treatment.

Indications and limitations for aged patients with chronic hepatitis C in pegylated interferon alfa-2b plus ribavirin combination therapy

BACKGROUND & AIMSnThis study investigated the efficacy and adverse effects of pegylated interferon (Peg-IFN) plus ribavirin therapy in aged patients with chronic hepatitis C (CH-C).nnnMETHODSnA total of 1040 naïve patients with CH-C (genotype 1, n=759; genotype 2, n=281), of whom 240 (23%) over 65 years old (y.o.), were treated with Peg-IFN alfa-2b plus ribavirin and assessed after being classified into five categories, according to age.nnnRESULTSnThe discontinuance rate was higher for patients over 70 y.o. (36%), the most common reason being anemia. In the presence of genotype 1, the SVR rate was similar (42-46%) among patients under 65 y.o. and declined (26-29%) among patients over 65 y.o. For patients over 65 y.o., being male (Odds ratio, OR, 3.5, p=0.035) and EVR (OR, 83.3, p<0.001) were significant factors for SVR, in multivariate analysis. The Peg-IFN dose was related to EVR, and when EVR was attained, 76-86% of patients over 65 y.o. achieved SVR. SVR was not achieved (0/35, 0/38, respectively) if a 1-log decrease and a 2-log decrease were not attained at week 4 and week 8, respectively. In the presence of genotype 2, the SVR rate was similar (70-71%) among patients under 70 y.o. and declined among patients over 70 y.o. (43%).nnnCONCLUSIONSnAged patients up to 65 y.o. with genotype 1 and 70 y.o. with genotype 2 can be candidates for Peg-IFN plus ribavirin therapy. The response-guided therapy can be applied for aged patients with genotype 1.

Involvement of transporter associated with antigen processing 2 (TAP2) gene polymorphisms in hepatitis C virus infection

BACKGROUND & AIMSnTransporter associated with antigen processing (TAP) has essential roles in the antigen-presenting systems, translocating antigenic peptides from the cytosol into the endoplasmic reticulum. The aim of this study was to clarify whether TAP polymorphisms are involved in hepatitis C virus (HCV) infection.nnnMETHODSnThe 145 HCV-infected Japanese patients examined in this study were categorized into two groups: 36 carriers with persistently normal alanine transaminase (ALT) values and 109 patients with chronic liver disease (CLD). TAP2 gene phenotypes were determined by means of polymerase chain reaction-restriction fragment length polymorphism, and their frequencies were compared between the two groups.nnnRESULTSnFrequencies of TAP2*0101, *0102, and *0201 were not different between the two groups. However, TAP2*0103 frequency in carriers with normal ALT levels was significantly higher than that in patients with CLD (44% vs. 16%; P = 0.00064, Pc < 0.005). Although the TAP2*0103 allele was tightly linked with class II DRB1*1302-DQB1*0604 haplotype in this study, the TAP2*0103 frequency in the normal ALT group was also significantly higher than that in the CLD group even in DRB1*1302-DQB1*0604-negative patients (31% vs. 10%; P = 0.0076, Pc < 0.05).nnnCONCLUSIONSnThese findings suggest that TAP2*0103 may be closely associated with low serum ALT activity in HCV-infected Japanese patients.