Functional Measurement of Hepatitis C Virus Core-Specific Cd8+T-Cell Responses in the Livers or Peripheral Blood of Patients by Using Autologous Peripheral Blood Mononuclear Cells as Targets or Stimulators
Resource
JOURNAL OF CLINICAL MICROBIOLOGY v.39 n.11 pp.3895-3901
Journal
JOURNAL OF CLINICAL MICROBIOLOGY
Journal Volume
v.39
Journal Issue
n.11
Pages
3895-3901
Date Issued
2001
Date
2001
Author(s)
FANG, SHIH-HUA
CHIANG, BOR-LUEN
WU, MEI-HUA
LAI, MING-YANG
YANG, PEI-MING
CHEN, DING-SHINN
HWANG, LIH-HWA
Abstract
As is widely recognized, CD8(+) cytotoxic T lymphocytes ( CTLs) play a crucial role in hepatitis C virus (HCV) infection, both in pathogenesis of liver injury and in clearing the virus. CTL studies with HCV-infected patients have been difficult because of the relatively low frequency of CTL precursors in the peripheral blood and because the targeted epitopes vary depending on the human leukocyte antigen (HLA) types of the individuals. This study attempts to overcome these problems by assessing the feasibility of using autologous peripheral blood mononuclear cells ( PBMCs) expressing viral antigens as stimulators or targets in order to monitor the CTL responses. Primary PBMCs were transduced using a retroviral vector pseudotyped with a vesicular stomatitis virus G glycoprotein expressing the HCV core gene . Additionally, the vector- transduced PBMCs were used as targets of CTL assays to measure the HCV core-specific CTL activities from the liver-infiltrating lymphocytes of six different HLA-type patients with chronic HCV infection. The core- expressing PBMCs also served as stimulators, allowing us to measure core- specific CD8+ T-cell responses by intracellular gamma interferon staining of the peripheral blood of hepatitis C patients who had received treatment with alpha interferon plus ribavirin. This approach provides an efficient means of measuring antigen-specific CTL responses without HLA constraints .
Subjects
VESICULAR STOMATITIS-VIRUS
CYTOTOXIC T-LYMPHOCYTES
MEDIATED GENE-TRANSFER
PSEUDOTYPED RETROVIRAL VECTORS
CRE RECOMBINASE
HIGH-TITER
SDGs
Type
journal article