Abstract
It is believed that the hepatitis C virus (HCV)-specific CD8+ cytotoxic T lymphocytes (CTLs) play a role in the development of liver cell injury and in the clearance of the virus. To develop a direct binding assay for HCV- specific CTLs, we generated two peptide-MHC tetramers by using the recombinant HLA A2.1 molecule and A2-restricted T cell epitopes of the HEW NS3 protein. With these reagents we are able to detect specific CD8+ cells in the blood of 15 of 20 HLA-A2+, HCV-infected patients, at a frequency ranging from 0.01% to 1.2% of peripheral CD8+ T cells. Phenotypic analysis of these specific cells indicated that there is a significant variation in the expression of the CD45 isoforms and CD27 in different patients. A 6-hour incubation of one patient's blood with NS3 peptides resulted in the activation of the epitope-specific CD8+ cells, as indicated by their expression of CD69 and IFN-γ. We also detected NS3-specific CD8+ T cells in the intrahepatic lymphocyte population isolated from liver biopsies of two HCV-infected patients. The frequency of these specific CD8+ cells in the liver was 1-2%, at least 30-fold higher than in the peripheral blood. All of the intrahepatic NS3-specific CD8+ T cells were CD69+, suggesting that they were activated CTLs. Direct quantitation and characterization of HCV-specific CTLs should extend our understanding of the immunopathogenesis and the mechanism of clearance or persistence of HCV.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 5692-5697 |
| Number of pages | 6 |
| Journal | Proceedings of the National Academy of Sciences of the United States of America |
| Volume | 96 |
| Issue number | 10 |
| DOIs | |
| State | Published - May 11 1999 |
| Externally published | Yes |
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ASJC Scopus subject areas
- Genetics
- General
Cite this
Quantitative analysis of hepatitis C virus-specific CD8+ T cells in peripheral blood and liver using peptide-MHC tetramers. / He, Xiaosong; Rehermann, Barbara; López-Labrador, F. Xavier; Boisvert, Judie; Cheung, Ramsey; Mumm, John; Wedemeyer, Heiner; Berenguer, Marina; Wright, Teresa L.; Davis, Mark M.; Greenberg, Harry B.
In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 96, No. 10, 11.05.1999, p. 5692-5697.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Quantitative analysis of hepatitis C virus-specific CD8+ T cells in peripheral blood and liver using peptide-MHC tetramers
AU - He, Xiaosong
AU - Rehermann, Barbara
AU - López-Labrador, F. Xavier
AU - Boisvert, Judie
AU - Cheung, Ramsey
AU - Mumm, John
AU - Wedemeyer, Heiner
AU - Berenguer, Marina
AU - Wright, Teresa L.
AU - Davis, Mark M.
AU - Greenberg, Harry B.
PY - 1999/5/11
Y1 - 1999/5/11
N2 - It is believed that the hepatitis C virus (HCV)-specific CD8+ cytotoxic T lymphocytes (CTLs) play a role in the development of liver cell injury and in the clearance of the virus. To develop a direct binding assay for HCV- specific CTLs, we generated two peptide-MHC tetramers by using the recombinant HLA A2.1 molecule and A2-restricted T cell epitopes of the HEW NS3 protein. With these reagents we are able to detect specific CD8+ cells in the blood of 15 of 20 HLA-A2+, HCV-infected patients, at a frequency ranging from 0.01% to 1.2% of peripheral CD8+ T cells. Phenotypic analysis of these specific cells indicated that there is a significant variation in the expression of the CD45 isoforms and CD27 in different patients. A 6-hour incubation of one patient's blood with NS3 peptides resulted in the activation of the epitope-specific CD8+ cells, as indicated by their expression of CD69 and IFN-γ. We also detected NS3-specific CD8+ T cells in the intrahepatic lymphocyte population isolated from liver biopsies of two HCV-infected patients. The frequency of these specific CD8+ cells in the liver was 1-2%, at least 30-fold higher than in the peripheral blood. All of the intrahepatic NS3-specific CD8+ T cells were CD69+, suggesting that they were activated CTLs. Direct quantitation and characterization of HCV-specific CTLs should extend our understanding of the immunopathogenesis and the mechanism of clearance or persistence of HCV.
AB - It is believed that the hepatitis C virus (HCV)-specific CD8+ cytotoxic T lymphocytes (CTLs) play a role in the development of liver cell injury and in the clearance of the virus. To develop a direct binding assay for HCV- specific CTLs, we generated two peptide-MHC tetramers by using the recombinant HLA A2.1 molecule and A2-restricted T cell epitopes of the HEW NS3 protein. With these reagents we are able to detect specific CD8+ cells in the blood of 15 of 20 HLA-A2+, HCV-infected patients, at a frequency ranging from 0.01% to 1.2% of peripheral CD8+ T cells. Phenotypic analysis of these specific cells indicated that there is a significant variation in the expression of the CD45 isoforms and CD27 in different patients. A 6-hour incubation of one patient's blood with NS3 peptides resulted in the activation of the epitope-specific CD8+ cells, as indicated by their expression of CD69 and IFN-γ. We also detected NS3-specific CD8+ T cells in the intrahepatic lymphocyte population isolated from liver biopsies of two HCV-infected patients. The frequency of these specific CD8+ cells in the liver was 1-2%, at least 30-fold higher than in the peripheral blood. All of the intrahepatic NS3-specific CD8+ T cells were CD69+, suggesting that they were activated CTLs. Direct quantitation and characterization of HCV-specific CTLs should extend our understanding of the immunopathogenesis and the mechanism of clearance or persistence of HCV.
UR - http://www.scopus.com/inward/record.url?scp=13044313467&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=13044313467&partnerID=8YFLogxK
U2 - 10.1073/pnas.96.10.5692
DO - 10.1073/pnas.96.10.5692
M3 - Article
C2 - 10318946
AN - SCOPUS:13044313467
VL - 96
SP - 5692
EP - 5697
JO - Proceedings of the National Academy of Sciences of the United States of America
JF - Proceedings of the National Academy of Sciences of the United States of America
SN - 0027-8424
IS - 10
ER -