Use of laser capture microdissection to map hepatitis C virus-positive hepatocytes in human liver

Abraham Kandathil, Frederik Graw, Jeffrey Quinn, Hyon S. Hwang, Michael Torbenson, Alan S. Perelson, Stuart Campbell Ray, David L Thomas, Ruy M. Ribeiro, Ashwin Balagopal

Research output: Contribution to journalArticle

Abstract

Background & Aims Hepatitis C virus (HCV) predominantly infects hepatocytes, but many hepatocytes are not infected; studies have shown that HCV antigens cluster within the liver. We investigated spatial distribution and determinants of HCV replication in human liver samples. Methods We analyzed liver samples from 4 patients with chronic HCV infection (genotype 1, Metavir scores 0-1) to estimate the proportion of infected hepatocytes and the amount of HCV viral RNA (vRNA) per cell. Single-cell laser capture microdissection was used to capture more than 1000 hepatocytes in grids, to preserve geometric relationships. HCV vRNA and interferon-induced transmembrane protein 3 (IFITM3) messenger RNA (the transcript of an interferon-stimulated gene) were measured in the same hepatocytes by quantitative polymerase chain reaction and assembled in maps to identify areas of high and low HCV replication. Results Patients' serum levels of HCV RNA ranged from 6.87 to 7.40 log10 IU/mL; the proportion of HCV-infected hepatocytes per person ranged from 21% to 45%, and the level of vRNA ranged from 1 to 50 IU/hepatocyte. Infection was not random; we identified clustering of HCV-positive hepatocytes using infected-neighbor analysis (P

Original languageEnglish (US)
JournalGastroenterology
Volume145
Issue number6
DOIs
StatePublished - Dec 2013

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Laser Capture Microdissection
Hepacivirus
Hepatocytes
Liver
Viral RNA
Virus Replication
Interferons
Hepatitis C Antigens
Chronic Hepatitis C
Virus Diseases
Cluster Analysis
Genotype
RNA

Keywords

  • Intrahepatic Infection
  • ISG
  • scLCM
  • Virology

ASJC Scopus subject areas

  • Gastroenterology

Cite this

Use of laser capture microdissection to map hepatitis C virus-positive hepatocytes in human liver. / Kandathil, Abraham; Graw, Frederik; Quinn, Jeffrey; Hwang, Hyon S.; Torbenson, Michael; Perelson, Alan S.; Ray, Stuart Campbell; Thomas, David L; Ribeiro, Ruy M.; Balagopal, Ashwin.

In: Gastroenterology, Vol. 145, No. 6, 12.2013.

Research output: Contribution to journalArticle

Kandathil, Abraham ; Graw, Frederik ; Quinn, Jeffrey ; Hwang, Hyon S. ; Torbenson, Michael ; Perelson, Alan S. ; Ray, Stuart Campbell ; Thomas, David L ; Ribeiro, Ruy M. ; Balagopal, Ashwin. / Use of laser capture microdissection to map hepatitis C virus-positive hepatocytes in human liver. In: Gastroenterology. 2013 ; Vol. 145, No. 6.
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