Hepatitis B surface antigen reduction by switching from long-term nucleoside/nucleotide analogue administration to pegylated interferon.

Hepatitis B surface antigen (HBsAg) reduction during nucleoside/nucleotide analogue (NA) therapy is slow and an alternative strategy for patients receiving ongoing NA to facilitate HBsAg reduction is required. We investigated whether switching to pegylated interferon (PEG-IFN) after long-term NA administration enhances HBsAg reduction. Forty-nine patients who switched from long-term NA to 48 weeks of PEG-IFN alfa-2a were studied. The mean duration of previous NA was 48 months (sequential group). A total of 147 patients who continued NA and matched for baseline characteristics were analysed for comparison (NA continuation group). The treatment response was defined as HBsAg reduction ≥1.0 logIU/mL at the end of PEG-IFN. HBsAg reduction at week 48 was 0.81±1.1 logIU/mL in the sequential group, which was significantly higher than that in the NA continuation group (0.11±0.3 logIU/mL, P < .001). The treatment response was achieved in 29% and 2% of the sequential group and NA continuation group (P < .001), and the odds ratio of sequential therapy for the treatment response was 19 compared with the NA continuation (P < .001). In patients tested positive for hepatitis B e antigen (HBeAg), HBeAg seroconversion was higher in the sequential group (44% vs 8%, P < .001). In HBeAg-negative patients, only patients in the sequential group achieved HBsAg loss. No patient needed to resume NA administration because of HBV DNA increase accompanied by alanine aminotransferase flares. In summary, sequential therapy with PEG-IFN after long-term NA enhances the reduction of HBsAg and may represent a treatment option to promote HBsAg loss.

Importance of cytotoxic T lymphocytes in the rejection of transplanted hepatocellular carcinoma.

Fischer rats became resistant to syngeneic hepatocellular carcinoma (FAA-HTC1) cells on repeated sensitization with mitomycin C-treated FAA-HTC1 cells. In contrast, FAA-HTC1 cells injected into the liver killed normal control Fischer rats within 2 months. Histopathological studies revealed massive accumulation of mononuclear cells in the tumor tissues of sensitized rats that rejected syngeneic FAA-HTC1 cells, whereas very few mononuclear cells were found in the tumor tissues of control rats. Cell populations infiltrating the tumor tissues were identified by flow cytometric analysis. Mononuclear cells found within the regressing tumors of the sensitized rats were identified as mostly T cells, and two-thirds of these T cells were CD8-positive. Compared with the activity in control rats, the killer activity of mononuclear cells infiltrating tumors was significantly increased in the sensitized rats 7 days after tumor inoculation. Depletion of CD8(+) T cells significantly reduced the cytotoxicity of mononuclear cells infiltrating tumors obtained from sensitized rats. In contrast, depletion of CD16(+) cells reduced the cytotoxicity of mononuclear cells infiltrating tumors obtained from both control and sensitized rats. Furthermore, the CD16(+) cell-depleted fraction of mononuclear cells infiltrating tumors showed significant cytotoxicity against FAA-HTC1 cells, but failed to show cytotoxicity against other syngeneic tumor cells or allogeneic hepatoma cells.

Hepatocellular carcinoma developed in a patient with chronic hepatitis C after the disappearance of hepatitis C virus due to interferon therapy.

A 62 year-old man was admitted to Asahikawa Medical College Hospital. Injection therapy of natural interferon-alpha was performed against chronic active hepatitis with hepatitis C virus infection. He successfully responded to interferon therapy with normalization of serum transaminases and disappearance of serum hepatitis C virus RNA. The liver function test remained within normal limits and serum hepatitis C virus RNA was not detected throughout the observation period. Three years later, CT examination demonstrated 2 small hepatic masses. Ultrasound-guided biopsy of the hepatic mass demonstrated well-differentiated hepatocellular carcinoma histologically. Laparoscopic examination revealed chronic hepatitis, but neither active inflammation nor cirrhotic changes were noted as an underlying liver disease. In the liver specimen, hepatitis C virus RNA was not detected by RT-PCR. Percutaneous ultrasound-guided ethanol injection therapy achieved complete necrosis of the hepatocellular carcinoma and there was no recurrence of hepatic cancer during the follow-up period. This case suggests that patients with chronic hepatitis C infection, who have complete disappearance of serum hepatitis C virus RNA by interferon therapy, should be followed-up carefully for the potential development of hepatocellular carcinoma.

Extensively calcified hemangioma of the diaphragm with increased 99mTc-hydroxymethylene diphosphonate uptake.

A 31-year-old woman visited an out-patient clinic, because of low-grade fever and general fatigue. She was referred to our hospital and admitted for examination of an abnormal shadow which had been found on the chest radiograph. She had experienced faint right lateral chest pain several times on the deep inspirations. Chest radiography showed a mass shadow with calcification in the right lower lung field on the mediastinal side. Chest radiographic computed tomography showed a 6x6 cm tumor in the right lung field. There were low-density areas with septae inside the tumor. Bone scintigraphy showed extremely high uptake of (99m)Tc-HMDP in the tumor. After surgical resection and pathological examination, we concluded that the tumor was an extensively calcified benign hemangioma of the diaphragm.

[Diagnosis of type C chronic hepatitis using PCR technology].

Many studies have suggested that examination of a patient's serum for viral RNA by reverse transcriptase-polymerase chain reaction (RT-PCR) is the most sensitive and specific serological means of determining chronic HCV infection. Moreover, technique to quantify HCV RNA in serum using PCR technology has been developed, and diagnosis of HCV genotype by PCR with mixed primers has been used for diagnosis of type C hepatitis. Herein, we discuss the laboratory technique and significance of various PCR methods. HCV RNA can be detected by amplification technique of RT-PCR. Primers in the 5'-noncoding region are frequently used to detect HCV RNA, because the RNA sequence in this region is the most conserved. We usually perform 30 cycles of amplification with outer primers and another 30 cycles with inner primers (nested RT-PCR). Using this technique, HCV RNA was detected in 96% of anti-HCV-positive chronic hepatitis patients. This PCR technique gives us important information for the diagnosis of type C chronic hepatitis indicating HCV infection. The competitive PCR technique is usually used to quantify HCV RNA. This method is based on complification of the target RNA with known amounts of synthetic mutant RNA. The mutant RNA should be amplified by the same primers for amplifying target RNA, and should be distinguished from target RNA by the size of the amplified DNA product by making a deletion or restriction site artificially. Quantifying HCV RNA before interferon therapy is useful to predict the effectiveness of the therapy; patients with a large amount of HCV RNA tend to have a poor outcome.(ABSTRACT TRUNCATED AT 250 WORDS)

[Histological findings of TTV-positive non-B non-C chronic hepatitis patients].

TT virus has been reported in association with patients with acute and chronic liver disease of unknown etiology. In order to estimate the pathogenesis of TTV, we investigated the liver histology in the patients of TTV-positive chronic hepatitis. The findings frequently observed in TTV-DNA positive cases were changes in liver parenchyma such as focal necrosis, hepatocyte degeneration or fatty change of various degree. On the other hand, degree of chronic inflammation in portal areas such as lymphocyte infiltration, piecemeal necrosis or fibrosis were relatively mild, which might suggest the good prognosis in TTV-positive cases. However, we could not find any differences in liver histology between TTV positive and negative patients.

[A case of type B chronic hepatitis showing rapid progression to liver cirrhosis with prolonged positive IgM anti-HBc].

We reported a case of type B chronic hepatitis with high titer of IgM anti-HBc continuously, which progressed to liver cirrhosis rapidly. The patient was a 37-years-old man, and was diagnosed as chronic type B hepatitis with severe activity by laparoscopy and liver biopsy. The titer of IgM anti-HBc persisted high level for more than 2 years. Then the second liver biopsy showed the progression to liver cirrhosis within only about 2 years. It is a very rare case to prolong positive IgM anti-HBc for such a long time. Anti-HBc production of PBMC was enhanced remarkably in this case. We considered that HBc antigens released into serum from hepatocytes with severe exacerbations had enhanced the anti-HBc producing activity of B lymphocytes. And the rapid progression in this case would be due to the episodes of severe exacerbations.

[Hepatic GOT and GPT activities in patients with various liver diseases--especially alcoholic liver disease].

GOT and GPT activities were measured in percutaneous needle biopsy specimens of human liver tissue from 98 cases including normal subjects and patients with various liver diseases. Hepatic GOT activity was markedly decreased in liver tissue of patients with nonalcoholic liver cirrhosis. Hepatic GPT activity was markedly decreased in liver tissue of patients with alcoholic liver cirrhosis. The GOT/GPT ratio in liver tissue was increased in patients with alcoholic liver cirrhosis (5.32 +/- 2.03) and alcoholic liver disease (4.78 +/- 2.43). The increased SGOT/SGPT ratio in patients with alcoholic liver disease is due to primarily to the increased LGOT/LGPT ratio.

[Clinical significance of HBeAg and anti-HBe in HBsAg positive liver diseases].

Variation of incidence of HBe antigen (HBeAg) and HBe antibody (anti-HBe) was examined by use of RIA in 72 patients with HBsAg positive liver diseases. 1) Percentage of positive HBeAg was highest (71.5%) in chronic active hepatitis with lobular distortion, followed by chronic active hapatitis without lobular distortion (70.0%) and acute hepatitis in asymptomatic HBsAg carriers (66.7%). In contrast, it was low in chronic inactive hepatitis (35.7%) and liver cirrhosis (38.5%). None of liver cancers showed HBeAg positive reaction. 2) Percentage of positive HBe antibody (anti-HBe) was highest in liver cancer (100%), followed by liver cirrhosis (61.5%) and chronic inactive hepatitis (50.0%). In acute hepatitis from asymptomatic HBsAg carriers no anti-HBe was found. In chronic active hepatitis the percentage of positive anti-HBe was low, 21.4 and 30.0% with and without lobular distortion, respectively. 3) In 45 patients with persistently positive HBsAg liver diseases, fluctuations of HBeAg and anti-HBe were followed over a period of one year in relation to serum GPT values, an indicator of clinical conditions. Serum GPT tended to fluctuate or to remain high in patients with persistently positive HBeAg or with sporadically positive HBeAg or anti-HBe, whereas it tended to become low or normal with persistently positive anti-HBe or with seroconversion from HBeAg to anti-HBe. However, there were some exceptions to this tendency. From these results we concluded that it is clinically of significant value to determine HBeAg and anti-HBe levels for the effective assessment of the activity and time course of HBsAg positive liver diseases.

Binding to R 1881 (methyltrienolone) of proteins from human benign prostatic hypertrophy.

Cytosol of human benign prostatic hypertrophy bound to R 1881 in a high affinity manner. Most of the protein which bound to R 1881 was recovered in the precipitate of a 0-30% saturation of ammonium sulfate, and was eluted in the void volume on a Sephadex G-200 column. The binding of cytosol to R 1881 was more inhibited by progestins than by dihydrotestosterone and estradiol-17 beta. The binder therefore seemed to be different from dihydrotestosterone-binding protein. The R 1881-binding component extracted from the nuclei by 0.4M KCl bound also to dihydrotestosterone in a high affinity manner. Cytosol prelabeled with R 1881 was bound to the nuclei in a nonsaturable process, and the extraction pattern of R 1881 by 0.4M KCl from the nuclei was almost identical to that in the case of dihydrotestosterone as the ligand. These results suggested that a part of the cytosolic protein which bound to R 1881 entered the nuclei where it bound to nuclear such components as dihydrotestosterone-binding protein.

Fas-mediated apoptosis of peripheral blood mononuclear cells in patients with hepatitis C.

In the present study, we demonstrated that a close relationship exists between hepatitis C virus (HCV) infection of peripheral blood mononuclear cells (PBMCs) and cell-surface Fas expression in patients with hepatitis C, and showed the possibility of PBMCs apoptosis via a Fas-mediated system. The expression of Fas on PBMCs was found by flowcytometric analysis to be significantly increased in these patients. In addition, the treatment of patients' PBMCs with anti-Fas antibody induced cell death, with nuclear condensation and fragmentation and cellular DNA fragmentation. These data indicate that the patients' PBMCs expressed a large amount of functional Fas on the cell surface and were susceptible to stimulation against Fas, causing apoptotic cell death. We then quantified the serum-soluble Fas ligand (sFasL), which was known to bind to Fas and induce the apoptotic signals into the sensitized cells. The patients' serum sFasL levels were significantly higher than those of normal subjects and showed a good negative correlation with their PBMC number. To demonstrate the correlation between Fas expression and HCV infection, nested reverse transcriptase polymerase chain reaction (RT-PCR) was performed to detect HCV RNA. Interestingly, HCV RNA was preferentially detected from Fas-positive cells but not from Fas-negative cells, which had been isolated from PBMCs by magnetic beads. These results suggest that HCV infection of PBMCs might induce Fas expression and additional stimulation such as sFasL might induce apoptosis in these Fas-expressing cells. These mechanisms, in addition to hypersplenism, may explain the decrease in the number of PBMCs observed in patients with chronic hepatitis C.