Production of antibody associated with non-A, non-B hepatitis in a chimpanzee lymphoblastoid cell line established by in vitro transformation with Epstein-Barr virus.

Shimizu, Yohko K.; Oomura, Masashi; Abe, Kengi; Uno, Masatune; Yamada, Erika; Ono, Yasushi; Shikata, Toshio

doi:10.1073/pnas.82.7.2138

Cited by 41 publications

(28 citation statements)

References 19 publications

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“…In 1985, p44 was shown to be a component of the doublewalled membranous tubules which appeared as a distinctive alteration in the cytoplasm of hepatocytes after intravenous administration of human non A, non B hepatitis inocula in chimpanzees (39,46,47). Neither the ultrastructural changes nor the expression of p44 was evident in chimpanzees experimentally infected with HBV.…”

Section: Discussionmentioning

confidence: 97%

Enhanced Expression of Interferon-Regulated Genes in the Liver of Patients with Chronic Hepatitis C Virus Infection: Detection by Suppression-Subtractive Hybridization

Patzwahl

Meier

Ramadori

et al. 2001

J Virol

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Section: Discussionmentioning

confidence: 97%

Enhanced Expression of Interferon-Regulated Genes in the Liver of Patients with Chronic Hepatitis C Virus Infection: Detection by Suppression-Subtractive Hybridization

Patzwahl

Meier

Ramadori

et al. 2001

J Virol

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“…All infected liver sections from chimpanzees were derived from experimental infection with NANBH agent strain F (no. 34, 60, 48 and 42) (Shimizu et aL, 1985), HDV (no. 92 and 48) (Shimizu et al, 1986), HBV or HAV at the glutamine-pyruvate transaminase peak during the acute phase.…”

Section: Methodsmentioning

confidence: 99%

“…Cultures showing positive IF were cloned by limiting dilution, rescreened using IF, and then expanded. These MAbs were then tested by IEM for reactivity with the intrahepatocytic microtubular aggregates according to the method described previously (Shimizu et al, 1985). Those antibodies capable of reacting with the denatured antigen polypeptide were finally screened using Western blot analysis.…”

Section: Methodsmentioning

confidence: 99%

“…In this way, we established two clonal cell lines, 48-1 and S-l, that produced class IgM MAbs recognizing hepatocytes from NANBH virus-infected chimpanzees (Shimizu et al, 1985). The 48-1 and S-1 MAbs reacted with liver biopsy specimens obtained from chimpanzees 0000-8415 O 1989 SGM with acute or chronic hepatitis caused by NANBH viruses, but not with biopsy tissues obtained from chimpanzees infected with either hepatitis A (HAV) or B (HBV) viruses, nor with material from normal chimpanzees.…”

Section: Introductionmentioning

confidence: 99%

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Production of Antibodies Directed against Microtubular Aggregates in Hepatocytes of Chimpanzees with Non-A, Non-B Hepatitis

Maeda

Honda²,

Hanawa³

et al. 1989

Journal of General Virology

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SUMMARYWe have previously used Epstein-Barr virus transformation to establish two clonal lymphoblastoid cell lines (48-1 and S-I) producing monoclonal antibodies against microtubular aggregates that appear in the hepatocytes of chimpanzees with non-A, non-B hepatitis (NANBH). To obtain additional antibodies directed against the same structure, the mouse hybridoma method was employed. Partially purified microtubular aggregates were prepared from liver homogenates of a chimpanzee with NANBH and used as the immunogen. Hybridoma cultures were first screened by radioimmunoassay against the partially purified antigen and secondly by immunofluorescence (IF) using liver sections from a chimpanzee with NANBH. Twenty-seven cultures exhibited positive IF reactions similar to those observed with the original antibodies, 48-1 and S-I, and were cloned by limiting dilution. The specificities of the monoclonal antibodies were tested by IF on liver biopsy specimens from chimpanzees with hepatitis A, B, D or NANBH and from normal chimpanzees. All the antibodies proved to be IgG. Immunoelectron microscopy revealed that all 27 antibodies bound to the same structure, the microtubular aggregates, in hepatocytes of chimpanzees with NANBH. To determine the size of the antigen polypeptide recognized by these antibodies, polyacrylamide gel electrophoresis and Western blot assays were performed. Nine of the 27 antibodies specifically reacted with a single polypeptide of Mr 44K (p44). The remaining 18 antibodies detected no antigen polypeptide on the filters. The anti-p44 antibodies were then tested using cross-competition assays with lzSI-labelled antibodies, and were found to be classifiable into three groups. In addition, the results indicate that at least three distinct epitopes are located on p44: epitope A recognized by group 1, epitope B recognized by group 2 and epitope C recognized by group 3.

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“…In 1985, before the discovery of the HCV gemone, we attempted to obtain antibodies associated with non-A, non-B hepatitis (now hepatitis C) by using the Epstein-Barr virus (EBV) transformation method (23). This method is based on the fact that B lymphocytes from immune individuals can be transformed in vitro by FM/ infection into lymphoblastoid cell lines capable of producing specific antibodies (28).…”

Section: Cytoplasmic Antigen 48-1mentioning

confidence: 99%

Ultrastructural Alterations and Expression of Cytoplasmic Antigen 48‐1 in Hepatocytes in Association with Hepatitis C Virus Infection

Shimizu

1992

Microbiology and Immunology

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Hepatitis type C is an important disease of humans ; it is responsible for more than 90% of the cases of posttransfusion non-A, non-B hepatitis (6,14). The etiologic agent of this disease, however, remained elusive for almost two decades, although the viral nature of the disease had been demonstrated already in 1978 by inoculating the serum or plasma from the patients into chimpanzees (1, 29). In 1989, the genome of the virus was finally discovered by the recombinant cDNA approach and the agent was termed hepatitis C virus (HCV) (5). HCV was revealed to contain a plus-strand RNA of about 10 kilonucleotides with one large open reading frame encoding a protein of 3010 amino acids (7, 13, 31) and comparative sequence analysis indicated that the virus is distantly related to the flaviviruses and pestiviruses (16). A sensitive assay for detecting HCV RNA by reverse transcription/polymerase chain reaction (RT/PCR) (32) and immunodiagnostic assays for antibodies against recombinant HCV proteins have been developed (14). The results from the serological assays indicated that most cases of hepatocellular carcinoma in Japan (19), Italy (8), and Spain (4) were associated with HCV infection.Before the emergence of these assays, chimpanzees (Pan troglodytes) were used for the characterization of the etiologic agent as the only reliable animal model for non-A, non-B hepatitis. The studies with chimpanzees have provided evidence that the virus is probably enveloped (9) and is approximately 30-60 nm in diameter (10). Additional research, however, has been hampered because of the limited availability of chimpanzees and the relatively low titer of the virus in clinical samples. To date, little is known about the replication and pathogenesis of this virus and the virion has not yet been visualized with certainty.Previously, my colleagues and I reported the presence of characteristic ultrastructural alterations (cytoplasmic tubular structures), detected by electron microscopy (EM) (22), and induction of a cytoplasmic antigen, detected by immunofluorescence staining (IF) with a monoclonal antibody (48-1) (23), in the hepatocytes of chimpanzees infected with non-A, non-B hepatitis virus (now HCV). These findings were initially thought to be specific for HCV infection, but were later found 911

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Production of antibody associated with non-A, non-B hepatitis in a chimpanzee lymphoblastoid cell line established by in vitro transformation with Epstein-Barr virus.

Cited by 41 publications

References 19 publications

Enhanced Expression of Interferon-Regulated Genes in the Liver of Patients with Chronic Hepatitis C Virus Infection: Detection by Suppression-Subtractive Hybridization

Enhanced Expression of Interferon-Regulated Genes in the Liver of Patients with Chronic Hepatitis C Virus Infection: Detection by Suppression-Subtractive Hybridization

Production of Antibodies Directed against Microtubular Aggregates in Hepatocytes of Chimpanzees with Non-A, Non-B Hepatitis

Ultrastructural Alterations and Expression of Cytoplasmic Antigen 48‐1 in Hepatocytes in Association with Hepatitis C Virus Infection

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