利用ELISPOT以及細胞內細胞泌素染色法來分析慢性B型肝炎病患之B型肝炎病毒特異性CD8陽性與CD4陽性的T細胞反應
Other Title
Analysis of Hepatitis B Virus-specific CD8 + and CD4 + T Cell
Responses in Patients Chronically Infected with Hepatitis B Virus
Using ELISPOT assay and Intracellular Cytokine Staining
Responses in Patients Chronically Infected with Hepatitis B Virus
Using ELISPOT assay and Intracellular Cytokine Staining
Date Issued
2001
Date
2001
Author(s)
陳健弘
DOI
892315B002037
Abstract
Taiwan is a hyperendemic area of
hepatitis B virus (HBV) infection. Around
15-20% adults in Taiwan are chronically
infected with HBV. Previous studies
demonstrated vigorous T cell responses to
HBV-encoded antigens developed in patients
with self-limited acute hepatitis B. In contrast,
weak or no T cell responses could be
detected in chronic hepatitis B patients.
The immunological assays used in these
previous studies are T cell proliferating assay
or cytokine ELISA assay in bulk cultures of
peripheral blood mononuclear cell (PBMC)
for CD4 + T cell responses, and chromium
release assays for CD8 + T cell responses.
These techniques suffer from the drawback
that they do not enable analysis of single cell
responses in the context of unselected
cellular background and need prolonged in
vitro restimulation. These methods may
underestimate the antigen-specific T cell
responses in chronic hepatitis B patients..
In contrast, ELISPOT assay and
intracellular cytokine staining are exquisitely
sensitive and can detect low frequency T
cells. They can detect the cytokine release
from an activated T cell and can be used for
enumeration of single cytokine-secreting
cells. These assays are believed to be at least
30-100 times more sensitive than the chromium release assay, and is efficient and
fast because it does not require any in vitro
cellular proliferation.
Therefore, in this current project, we
applied the ELISPOT assays and intracellular
cytokine staining to assess the HBV
antigen-specific CD4 + and CD8 + T cell
responses in patients infected with HBV. We
successfully establish the ELISPOT and
intracellular cytokine staining assays.
However, only weak or no T cell responses
was detected in patients with chronic HBV
infection. This implied that T cell
hypo-responsiveness may play an important
role in the pathogenesis of chronic hepatitis
B. Thus, enhancing the HBV-specific T cell
responses would be one of the therapeutic targeting for chronic hepatitis B.
Subjects
hepatitis B virus
antigen-specific
T cell responses
T cell responses
intracellular cytokine staining
CD4+
CD8+
SDGs
Publisher
臺北市：國立臺灣大學醫學院內科
Type
report
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