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Ultra-structural localisation of hepatocellular PKR protein using immuno-gold labelling in chronic hepatitis C virus disease

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Abstract

The greater resistance of HCV genotype 1 infection to IFN therapy has been partially attributed to functional inhibition of the type 1 interferon induced anti-viral protein PKR in vitro. Whether PKR has antiviral activity against HCV in vivo is unknown. Whilst the ultra-structural localisation of PKR is known in vitro, it is not defined in chronic hepatitis C disease. Using a novel immuno-gold technique we characterised the expression of intrahepatic PKR protein at the ultra-structural level in four patients with chronic HCV disease compared to normal human PBMCs, HepG2 cells and a normal human liver biopsy. All four HCV patients labelled for PKR protein, localising to the nucleus, nucleolus and cytoplasm. Nuclear labelling was confined mainly to the nucleolus and euchromatin. Cytoplasmic labelling was evident within smooth vesicles. Strong immunogold labelling was also evident within the cisternae of the rough endoplasmic reticulum. A similar pattern of ultra-structural nuclear and cytoplasmic PKR protein labelling was seen in PBMCs from healthy donors, HepG2 cells and a normal liver biopsy. The mean nuclear and cytoplasmic count for PKR protein in the HCV group was 21 ± 4 and 18 ± 3 gold particles/μm2, respectively. This represented an increase, though not statistically significant, in nuclear and cytoplasmic labelling for PKR protein in HCV biopsies relative to normal liver tissue.

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Acknowledgments

Dr. MacQuillan was supported by a Postgraduate Research Scholarship from the National Health and Medical Research Council of Australia, an Athelstan and Amy Saw Medical Research Fellowship from the University of Western Australia and a grant from the Sir Charles Gairdner Hospital Research Fund.

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Authors and Affiliations

  1. Department of Gastroenterology and Hepatology, Sir Charles Gairdner Hospital, Nedlands, WA, Australia

    Gerry C. MacQuillan & Gary P. Jeffrey

  2. School of Medicine and Pharmacology, University of Western Australia, Nedlands, WA, Australia

    Gerry C. MacQuillan, Jane E. Allan & Gary P. Jeffrey

  3. Department of Anatomical Pathology, PathWest, QEII Medical Centre, Nedlands, WA, Australia

    Paul Caterina, Bastiaan de Boer & Michael A. Platten

  4. Hollywood Private Hospital, Nedlands, WA, Australia

    William D. Reed

Authors
  1. Gerry C. MacQuillan
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  2. Paul Caterina
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  3. Bastiaan de Boer
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  4. Jane E. Allan
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  5. Michael A. Platten
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  6. William D. Reed
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  7. Gary P. Jeffrey
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Correspondence to Gerry C. MacQuillan.

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MacQuillan, G.C., Caterina, P., de Boer, B. et al. Ultra-structural localisation of hepatocellular PKR protein using immuno-gold labelling in chronic hepatitis C virus disease. J Mol Hist 40, 171–176 (2009). https://doi.org/10.1007/s10735-009-9227-0

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  • DOI: https://doi.org/10.1007/s10735-009-9227-0

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