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Journal of Virology, November 2008, p. 10487-10492, Vol. 82, No. 21
0022-538X/08/$08.00+0     doi:10.1128/JVI.00588-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Kinetic Analysis by Real-Time PCR of Hepatitis C Virus (HCV)-Specific T Cells in Peripheral Blood and Liver after Challenge with HCV{triangledown}

Ramesh K. Ramalingam,1 Dirk Meyer-Olson,2 Naglaa H. Shoukry,3 David G. Bowen,4 Christopher M. Walker,4,5 and Spyros A. Kalams1,6*

Department of Medicine, Division of Infectious Diseases, Vanderbilt University Medical Center, Nashville, Tennessee 37232,1 The Abteilung Klinische Immunologie, Medizinische Hochschule Hannover, Hanover, Germany,2 Department of Medicine, University of Montreal and University of Montreal Hospital Research Centre (CRCHUM), Montreal, Canada,3 The Research Institute at Nationwide Children's Hospital, Columbus, Ohio 43205,4 Department of Pediatrics, College of Medicine and Public Health, The Ohio State University, Columbus, Ohio 43205,5 Department of Microbiology and Immunology, Vanderbilt University Medical Center, Nashville, Tennessee 372326

Received 16 March 2008/ Accepted 8 August 2008

Intrahepatic virus-specific CD8+ T cells are thought to be important for the control of hepatitis C virus (HCV) infection, yet the precise kinetics for the expansion of epitope-specific T cells over the course of infection are difficult to determine with currently available methods. We used a real-time PCR assay to measure the frequency of clonotypic HCV-specific CD8+ T cells in peripheral blood and snap-frozen liver biopsy specimens of two chimpanzees (Pan troglodytes) with previously resolved HCV infection who were rechallenged with HCV. In response to rechallenge, the magnitude of each clonotypic response was 10-fold higher in the liver than in the blood, and the peak clonotype frequency was concurrent with the peak viral load. The higher frequency of HCV-specific clonotypes in the liver than in peripheral blood was maintained for at least 3 months after the clearance of viremia. After antibody-mediated CD8+ T-cell depletion and another viral challenge, the rebound of these clonotypes was seen prior to an appreciable reconstitution of CD8+ T-cell values and, again, at higher frequencies in the liver than in peripheral blood. These data demonstrate the importance of intrahepatic virus-specific CD8+ T cells for the clearance of infection and the rapid kinetics of expansion after virus challenge.

* Corresponding author. Mailing address: Infectious Diseases Unit, Dept. of Internal Medicine and Dept. of Microbiology and Immunology, Vanderbilt University Medical Center, MCN A2205b, Nashville, TN 37232. Phone: (615) 322-2035. Fax: (615) 343-6160. E-mail: spyros.a.kalams{at}vanderbilt.edu

{triangledown} Published ahead of print on 20 August 2008.

Journal of Virology, November 2008, p. 10487-10492, Vol. 82, No. 21
0022-538X/08/$08.00+0     doi:10.1128/JVI.00588-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.





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