Expression of processed envelope protein of hepatitis C virus in mammalian and insect cells.

The putative envelope protein of hepatitis C virus (HCV) was expressed in insect cells by using a baculovirus expression vector and in monkey COS cells under the control of exogenous promoters. The expressed envelope proteins, identified by immunoblot analysis using sera from patients with chronic HCV infection, were a series of glycoproteins of 35 to 24 kDa (gp35-24) in insect cells and a single species of glycoprotein of 35 kDa (gp35) in monkey cells. The size difference of these proteins was due to the different degrees of glycosylation.

Seroprevalence of hepatitis C virus nucleocapsid antibodies in patients with cryptogenic chronic liver disease.

The serological responses to two different hepatitis C virus antigens were studied by enzyme-linked immunosorbent assay in a variety of chronic liver diseases and in healthy blood donors. The study population comprised 97 cases of cryptogenic chronic liver disease (40% with a history suggestive of parenterally transmitted non-A, non-B hepatitis and 60% without such a history), 87 cases of other well-characterized chronic liver diseases and 96 voluntary blood donors. The commercially available C100-3 assay and a new assay utilizing a 22 kD recombinant protein (c22) from the nucleocapsid region of the virus were used for antibody detection.

[Relationship between Ga-67 uptake and radiotherapeutic response of primary lung cancer (squamous cell carcinoma)].

This investigation was undertaken to evaluate the relationship between Ga-67 uptake and radiotherapeutic response to primary lung cancer (squamous cell carcinoma), Ga-67 uptake of tumor was estimated on 16 patients with untreated primary lung cancer (squamous cell carcinoma). Ga-67 uptake was then compared with the response to radiation therapy (tumor reduction ratio). There was statistically significant inverse correlation between Ga-67 uptake and response to radiation therapy (r = -0.

Correlation between detection of anti-viral antibody and histopathological disease activity in an epidemic of hepatitis C.

There was an epidemic of non-A non-B hepatitis in a small area of a town in the central part of Japan, which began with an outbreak of several patients in 1981 and then spread extensively with the result that about one third of the inhabitants showed abnormality in serum liver function tests at the health check performed in 1985. We determined histological diagnoses on that occasion for 167 individuals of the abnormal population and recently assayed antibodies against hepatitis C virus (HCV) for most of their sera left available. Histologically, chronic active hepatitis (CAH) was the major pattern, accounting for 59.

Discrimination of hepatitis C virus in liver tissues from different patients with hepatocellular carcinomas by direct nucleotide sequencing of amplified cDNA of the viral genome.

We investigated the presence of the hepatitis C virus (HCV) genome in liver tissues of eight different patients with hepatocellular carcinoma by using the reverse transcriptase polymerase chain reaction (PCR) method. RNA was extracted separately from cancerous and peripheral noncancerous portions of the liver tissues of each patient. For reverse transcriptase PCR, we used sets of primers derived either from nonstructural region 3 (the NS3 region) or from the nucleocapsid-envelope (C/E) region of the HCV genome.

Intrafamilial transmission of hepatitis C virus in Japan.

To study the intrafamilial transmission of hepatitis C virus (HCV), 36 family members of 16 patients with anti-HCV (anti-C100-3)-positive chronic liver disease were screened for anti-HCV by an enzyme-linked immunosorbent assay (ELISA). Clusters of anti-HCV-positive individuals were observed in 2 of 16 families (12.5%).

The particle size of hepatitis C virus estimated by filtration through microporous regenerated cellulose fibre.

To estimate the particle size of hepatitis C virus (HCV), a major causative agent of post-transfusion non-A, non-B hepatitis, we filtered plasma or serum samples through microporous cellulose fibres with different pore sizes. The amount of HCV particles in samples before and after filtration was determined by a quantitative reverse transcriptase polymerase chain reaction (PCR) method. Since there is no quantitative biological assay for HCV, except for that in chimpanzees, the HCV titre obtained from the PCR method was used in an equation constructed previously for application to filtration experiments with a flavivirus which is distantly related to HCV.

Molecular cloning of cDNA of hepatitis C virus (HCV) genome from a healthy carrier in an aboriginal community in Taiwan with high prevalence of HCV infection.

We analyzed hepatitis C virus (HCV) obtained from a healthy HCV carrier in an aboriginal community with high prevalence of HCV infection. By use of random primers and specific oligonucleotide primers, a cDNA fragment of putative non-structural 3 (NS3) region of the HCV genome was cloned by reverse transcription-polymerase chain reaction (RT-PCR) using RNA extracted from a serum sample of a healthy HCV carrier in Taur-Yuan village. The nucleotide and deduced amino acid sequences from the 583 nucleotides long cDNA were compared with those of equivalent region of HCV of other previously reported clones: [J1, HCV-J (regarded as major type of HCV in Japan) and US prototype].

Preferential expression of the large hepatitis B virus surface antigen gene by an adenovirus-hepatitis B virus recombinant.

Using an adenovirus-hepatitis B virus (HBV) recombinant, expression of the HBV surface antigen (HBsAg) genes was examined in various cell lines using S1 nuclease mapping and radioimmunoassay. The steady-state level of the 2.4 kb RNA encoding the large HBsAg was much greater than, or the same as, that of the 2.

Serodiagnosis of hepatitis C virus (HCV) infection with an HCV core protein molecularly expressed by a recombinant baculovirus.

An enzyme-linked immunosorbent assay (ELISA) was developed for serological diagnosis of hepatitis C virus (HCV) infection, using HCV core protein (p22) synthesized by a recombinant baculovirus. Among 58 clinically well-defined chronic non-A, non-B hepatitis (NANBH) patients, 49 (84.5%) were positive for p22 antibody (anti-p22), whereas 42 (72.

Expression of processed core protein of hepatitis C virus in mammalian cells.

A structural protein of hepatitis C virus (HCV) was expressed in monkey COS cells under the control of an exogenous promoter, and a protein of 22 kDa was identified by immunoblot analysis. This protein (p22), which was produced by processing in COS cells, reacted specifically to sera of chronic hepatitis C patients, and its coding region was mapped at the most amino-terminal part of the HCV polyprotein. These results suggested that the p22 protein is the nucleocapsid (core) protein of HCV.

Prevalence of antibody against the core protein of hepatitis C virus in patients with hepatocellular carcinoma.

We have cloned the whole structural region of the hepatitis C virus (HCV) genome and transiently expressed the nucleocapsid protein in animal cells. Since the nucleotide sequences of this region of the HCV genome has been shown to be highly conserved among different HCV isolates, the assay detecting the antibody to this expressed protein is useful for studying the pathogenicity of HCV. In this work, we investigated the presence of antibodies to HCV nucleocapsid protein (p22) in patients with hepatocellular carcinoma (HCC) and compared its frequency with that of antibody to HCV non-structural protein (C-100), which is presently applied for blood screening for transfusion and diagnosis for chronic hepatitis C.

HBx gene of hepatitis B virus induces liver cancer in transgenic mice.

The exact role of hepatitis B virus in the development of liver cancer is not known. The recent identification of a viral regulatory gene HBx suggests a possible direct involvement of the virus whereby the HBx protein, acting as a transcriptional transactivator of viral genes, may alter host gene expression and lead to the development of hepatocellular carcinoma. We have tested this possibility of placing the entire HBx gene under its own regulatory elements directly into the germline of mice.

Detection of hepatitis C virus antibody in patients with autoimmune hepatitis and other chronic liver diseases.

To clarify the relationship between autoimmune hepatitis (AIH) and the hepatitis C virus (HCV), we investigated the prevalence of antibodies to HCV (anti-HCV) by an enzyme-linked immunosorbent assay in patients with AIH, primary biliary cirrhosis (PBC), rheumatoid arthritis and multiple myeloma. The antibody was detected in 9 out of 18 patients with AIH (50%), in 3 out of 23 with PBC (23%), in 2 out of 10 with rheumatoid arthritis (20%), and in 5 out of 9 with multiple myeloma (56%). However, the optical density values in these patients were lower than those observed in non-A, non-B hepatitis (NANBH).

Efficient cloning of hepatitis B virus DNA from single-stranded replicative intermediates and its application to S1 mapping of viral RNA in human liver tissue.

An efficient method for cloning subgenomic fragments of the hepatitis B virus (HBV) was developed, utilizing its abundant single-stranded replicative intermediates. The total genomic DNA obtained from the liver tissue of patients with chronic HBV infection was treated by using the Klenow fragment of E. coli DNA polymerase I without adding any exogenous primers.

Alternative splicing of hepatitis B virus RNAs in HepG2 cells transfected with the viral DNA.

We identified a novel spliced RNA of 2.6 kb from a human hepatoma cell line HepG2 transfected with the hepatitis B virus (HBV) genome. The splicing acceptor site of the novel 2.

The putative nucleocapsid and envelope protein genes of hepatitis C virus determined by comparison of the nucleotide sequences of two isolates derived from an experimentally infected chimpanzee and healthy human carriers.

cDNA fragments of a 5'-terminal region of the hepatitis C virus (HCV) genome were isolated by the reverse polymerase chain reaction from RNA extracted from plasma samples of healthy Japanese carriers. Their nucleotide sequence was compared with that of the original isolate which had been passaged twice in chimpanzees. No deletions or insertions were observed between the two sequences in the regions examined.

Allele loss on chromosome 16 associated with progression of human hepatocellular carcinoma.

Loss of heterozygosity on chromosome 16 is a common genetic alteration in human hepatocellular carcinoma (HCC). To clarify the pathogenetic significance of allele loss on chromosome 16, we performed restriction fragment length polymorphism analysis of 70 surgically resected tumors by using 15 polymorphic DNA markers for chromosome 16. Loss of heterozygosity on chromosome 16 was detected in 36 (52%) of 69 informative cases, and the common region of allele loss in these 36 tumors was located between the HP locus (16q22.

Hepatitis C virus infection is associated with the development of hepatocellular carcinoma.

A possible causative role for the recently discovered hepatitis C virus (HCV) in the development of hepatocellular carcinoma (HCC) was investigated by assay of sera from HCC patients in Japan for antibodies to a recombinant HCV antigen and to hepatitis B virus (HBV) antigens. Among the 253 HCC patients examined, 156 (61.7%) had no serum markers of either a previous or a current HBV infection (group I), 46 (18.

Detection of hepatitis C virus cDNA sequence by the polymerase chain reaction in hepatocellular carcinoma tissues.

We found the presence of hepatitis C virus (HCV) infection in liver tissues of hepatocellular carcinoma (HCC) patients who had antibodies to HCV but no serological markers for hepatitis B virus infection by the sensitive reverse transcription/polymerase chain reaction (R/PCR) method. The primers used were derived from the non-structural (NS) 3 and/or the structural (C/E) region. Amplified cDNA sequences of HCV were detected in either cancerous or non-cancerous portion of liver tissues from four out of eight HCC patients with primers of NS3 region.