Chronic hepatitis in chimpanzee carriers of hepatitis B virus: morphologic, immunologic, and viral DNA studies.

Shouval, Daniel; Chakraborty, Parthasarathi; Ruiz-Opazo, Nelson; Baum, Steven K.; Spigland, Ilya; Muchmore, Elizabeth; Gerber, Michael A.; Thung, Swan N.; Pópper, Hans; Shafritz, David A.

doi:10.1073/pnas.77.10.6147

Cited by 42 publications

(22 citation statements)

References 20 publications

(14 reference statements)

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“…The presence of slowly migrating closed circular DNA in human livers infected with HBV has been observed (13). The pattern of electrophoretic migration of DNA bands seen in the present study may be comparable to the pattern reported for liver and pancreatic tissues infected with DHBV (10 (14) by digestion with Bgl II endonuclease.…”

supporting

confidence: 75%

Hepatitis B virus DNA in Kaposi sarcoma.

Siddiqui¹

1983

Proc. Natl. Acad. Sci. U.S.A.

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(14) by digestion with Bgl II endonuclease. The pSH6 DNA was prepared by standard methods (14). The HBV DNA fragment was purified from the excised gel piece by dissolution in sodium iodide solution, followed by addition of glass powder beads to bind the DNA (15). The DNA was eluted in 0.05 M Tris/0.001 M EDTA, pH 7.5 M. The isolated 2.8-kilobase Bgl II fragment, which spans about 80% of the HBV genome, includes a complete hepatitis B surface antigen (HBsAg) gene and most of hepatitis B core antigen (HBcAg) gene. The fragment was radiolabeled in vitro by the method of nick-translation (16). The specific activity of the labeled DNA was 1 X 108 cpm/ig. [a-32P]dCTP (800 Ci/mmol; 1 Ci = 3.7 X 1010 Bq) was purchased from New England Nuclear. Escherichia coli DNA polymerase I was from Bethesda Research Laboratories. Nitrocellulose sheets (BA85) (0.45-,tm pore size) were obtained from Schleicher & Schuell.Dane particles were purified by centrifugation of the hepatitis serum in an SW 27 rotor through a 20% sucrose solution for 16 hr at 25,000 rpm. HBV DNA was radiolabeled by an endogenous DNA polymerase reaction as described (9). After the reaction, DNA was extracted with phenol by standard methods.Gel Electrophoresis. The Kaposi sarcoma DNA was digested with appropriate restriction endonucleases and fractionated in a 0.75% agarose gel at 60 V for 12-16 hr. Restriction enzymes were purchased from Bethesda Research Laboratories. Gels were stained with ethidium bromide at 5 ,ug/ml and visualized with a UV lamp transilluminator.Southern Hybridization. The gels were treated with 0.5 M NaOH and 1.5 M NaCl for 1 hr with shaking. Gels were transferred to 1.5 M NaCl/0.5 M Tris, pH 7.5, and washed twice for 30 min each. DNA fragments were transferred from agarose gels to nitrocellulose filters for 5-6 hr (17). Filters were then baked at 800C for 2 hr. The sheets were treated prior to hyAbbreviations: HBV, hepatitis B virus; DHBV, Duck hepatitis B virus; AIDS, acquired immunodeficiency syndrome; NaCl/Cit, 0.15 M NaCl/ 0.015 M sodium citrate, pH 7; bp, base pairs. 4861The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. §1734 solely to indicate this fact.

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supporting

confidence: 75%

Hepatitis B virus DNA in Kaposi sarcoma.

Siddiqui¹

1983

Proc. Natl. Acad. Sci. U.S.A.

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“…3). The data for the 10 12 vg group are shown in Fig. 3, but similar results were obtained in all groups.…”

Section: Construction Of Trans-splicing Aav Vectorssupporting

confidence: 62%

“…Chimpanzees are susceptible to HBV infection, but only develop a mild liver inflammatory reaction (10), and their use in laboratory research is further limited because of ethical and financial considerations. Studies of HBV-related viruses in woodchucks, ground squirrels and ducks have improved our knowledge of HBV virology and the development of antiviral agents, but have not led to a better understanding of HBV immunopathology (11).…”

Section: Introductionmentioning

confidence: 99%

A murine model of hepatitis B-associated hepatocellular carcinoma generated by adeno-associated virus-mediated gene delivery

et al. 2011

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Abstract.A relevant animal model is critical for investigating the pathogenic mechanisms underlying hepatitis B virus (HBV)-induced hepatocellular carcinoma (HCC). Mice are not naturally infected by HBV, presumably due to the lack of HBV receptors on mouse hepatocytes. To bypass this entry step of HBV infection, we report generation of a novel HBV model in immunocompetent mice by hepatic delivery of the HBV genome using trans-splicing adeno-associated viral vectors (AAV/HBV). We confirmed production of HBV virions and proteins in the liver and circulation in all AAV/ HBV-transduced mice in all four immunocompetent mouse strains tested. These mice produced antigen and antibody profiles similar to that observed in chronic HBV patients. Importantly, 12-16 months later, all 12 AAV/HBV-transduced mice developed macroscopically visible liver-tumor nodules. Ten of the twelve tumors were characterized with typical HCC features. This AAV/HBV-transduced murine HCC model provides a useful instrument for studying the pathogenesis of HBV-associated HCC and the development of HCC therapeutic interventions.

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“…However, there are important biomedical (as well as obvious morphological and cognitive) differences between the two species, which thus far have eluded any molecular explanation within this supposedly 1% diversity range. Among these are our differential handling of a number of infectious agents, e.g., HIV (progression to AIDS) (4), late complications of hepatitis B and C (5,6), as well as susceptibility to Plasmodium falciparum (7), which are of utmost public health importance. The molecular basis of these distinctive traits is thought to be in large part encoded within the MHC, where MHC class I molecules sample pathogen-derived antigenic peptides for recognition by the CD8 ϩ ␣␤ T cell receptor expressing cytotoxic T cells (8).…”

mentioning

confidence: 99%

Comparative sequencing of human and chimpanzee MHC class I regions unveils insertions/deletions as the major path to genomic divergence

Anzai

Shiina²,

Kimura³

et al. 2003

Proc. Natl. Acad. Sci. U.S.A.

118

108

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Despite their high degree of genomic similarity, reminiscent of their relatively recent separation from each other (Ϸ6 million years ago), the molecular basis of traits unique to humans vs. their closest relative, the chimpanzee, is largely unknown. This report describes a large-scale single-contig comparison between human and chimpanzee genomes via the sequence analysis of almost one-half of the immunologically critical MHC. This 1,750,601-bp stretch of DNA, which encompasses the entire class I along with the telomeric part of the MHC class III regions, corresponds to an orthologous 1,870,955 bp of the human HLA region. Sequence analysis confirms the existence of a high degree of sequence similarity between the two species. However, and importantly, this 98.6% sequence identity drops to only 86.7% taking into account the multiple insertions͞deletions (indels) dispersed throughout the region. This is functionally exemplified by a large deletion of 95 kb between the virtual locations of human MICA and MICB genes, which results in a single hybrid chimpanzee MIC gene, in a segment of the MHC genetically linked to species-specific handling of several viral infections (HIV͞SIV, hepatitis B and C) as well as susceptibility to various autoimmune diseases. Finally, if generalized, these data suggest that evolution may have used the mechanistically more drastic indels instead of the more subtle singlenucleotide substitutions for shaping the recently emerged primate species.

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Chronic hepatitis in chimpanzee carriers of hepatitis B virus: morphologic, immunologic, and viral DNA studies.

Cited by 42 publications

References 20 publications

Hepatitis B virus DNA in Kaposi sarcoma.

Hepatitis B virus DNA in Kaposi sarcoma.

A murine model of hepatitis B-associated hepatocellular carcinoma generated by adeno-associated virus-mediated gene delivery

Comparative sequencing of human and chimpanzee MHC class I regions unveils insertions/deletions as the major path to genomic divergence

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