Visualizing Hepatitis C Virus Infections in Human Liver by Two-Photon Microscopy

Yuqiong Liang, Tuya Shilagard, Shu Yuan Xiao, Ned Snyder, Daryl Lau, Luca Cicalese, Heidi Weiss, Gracie Vargas, Stanley M. Lemon

Research output: Contribution to journalArticle

140 Citations (Scopus)

Abstract

Background & Aims: Although hepatitis C virus (HCV) is a common cause of cirrhosis and liver cancer, efforts to understand the pathogenesis of HCV infection have been limited by the low abundance of viral proteins expressed within the liver, which hinders the detection of infected cells in situ. This study evaluated the ability of advanced optical imaging techniques to determine the extent and distribution of HCV-infected cells within the liver. Methods: We combined 2-photon microscopy with virus-specific, fluorescent, semiconductor quantum dot probes to determine the proportion of hepatocytes that were infected with virus in frozen sections of liver tissue obtained from patients with chronic HCV infection. Results: Viral core and nonstructural protein 3 antigens were detected readily in liver tissues from patients with chronic infection without confounding tissue autofluorescence. Specificity was confirmed by blocking with specific antibodies and by tissue colocalization of distinct viral antigens. Between 7% and 20% of hepatocytes were infected in patients with plasma viral RNA loads of 105 IU/mL or greater. Infected cells were in clusters, which suggested spread of the virus from cell to cell. Double-stranded RNA, a product of viral replication, was abundant within cells at the center of such clusters, but often scarce in cells at the periphery, consistent with more recent infection of cells at the periphery. Conclusions: Two-photon microscopy provides unprecedented sensitivity for the detection of HCV proteins and double-stranded RNA. Studies using this technology indicate that HCV infection is a dynamic process that involves a limited number of hepatocytes. HCV spread between cells is likely to be constrained by host responses.

Original languageEnglish
Pages (from-to)1448-1458
Number of pages11
JournalGastroenterology
Volume137
Issue number4
DOIs
StatePublished - Oct 2009

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Virus Diseases
Photons
Hepacivirus
Microscopy
Liver
Hepatocytes
Double-Stranded RNA
Viruses
Viral Nonstructural Proteins
Viral Core Proteins
Quantum Dots
Semiconductors
Viral Antigens
Optical Imaging
Viral RNA
Frozen Sections
Viral Proteins
Chronic Hepatitis C
Liver Neoplasms
Infection

ASJC Scopus subject areas

  • Gastroenterology

Cite this

Liang, Y., Shilagard, T., Xiao, S. Y., Snyder, N., Lau, D., Cicalese, L., ... Lemon, S. M. (2009). Visualizing Hepatitis C Virus Infections in Human Liver by Two-Photon Microscopy. Gastroenterology, 137(4), 1448-1458. https://doi.org/10.1053/j.gastro.2009.07.050

Visualizing Hepatitis C Virus Infections in Human Liver by Two-Photon Microscopy. / Liang, Yuqiong; Shilagard, Tuya; Xiao, Shu Yuan; Snyder, Ned; Lau, Daryl; Cicalese, Luca; Weiss, Heidi; Vargas, Gracie; Lemon, Stanley M.

In: Gastroenterology, Vol. 137, No. 4, 10.2009, p. 1448-1458.

Research output: Contribution to journalArticle

Liang, Y, Shilagard, T, Xiao, SY, Snyder, N, Lau, D, Cicalese, L, Weiss, H, Vargas, G & Lemon, SM 2009, 'Visualizing Hepatitis C Virus Infections in Human Liver by Two-Photon Microscopy', Gastroenterology, vol. 137, no. 4, pp. 1448-1458. https://doi.org/10.1053/j.gastro.2009.07.050
Liang, Yuqiong ; Shilagard, Tuya ; Xiao, Shu Yuan ; Snyder, Ned ; Lau, Daryl ; Cicalese, Luca ; Weiss, Heidi ; Vargas, Gracie ; Lemon, Stanley M. / Visualizing Hepatitis C Virus Infections in Human Liver by Two-Photon Microscopy. In: Gastroenterology. 2009 ; Vol. 137, No. 4. pp. 1448-1458.
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