HEPATITIS-B VIRUS (HBV)-SPECIFIC CYTOTOXIC T-CELL RESPONSE IN HUMANS - PRODUCTION OF TARGET-CELLS BY STABLE EXPRESSION OF HBV-ENCODED PROTEINS IN IMMORTALIZED HUMAN B-CELL LINES

被引:64
作者
GUILHOT, S
FOWLER, P
PORTILLO, G
MARGOLSKEE, RF
FERRARI, C
BERTOLETTI, A
CHISARI, FV
机构
[1] Scripps Res Inst, RES INST, DEPT MOLEC & EXPTL MED, LA JOLLA, CA 92037 USA
[2] ROCHE INST MOLEC BIOL, DEPT NEUROSCI, NUTLEY, NJ 07110 USA
[3] UNIV PARMA, DEPT INFECT DIS, I-43100 PARMA, ITALY
关键词
D O I
10.1128/JVI.66.5.2670-2678.1992
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
To analyze the hepatitis B virus (HBV)-specific cytotoxic T-cell (CTL) response during acute and chronic viral hepatitis, target cells that express HBV-encoded antigens in the context of the appropriate HLA restriction element must be available for each subject studied. Since HBV is not infectious for human cells in vitro, such target cells must be produced by DNA-mediated gene transfer into cultured human primary cells or cell lines. For this purpose, we have developed a panel of Epstein-Barr virus-based episomal expression vectors containing each of the HBV open reading frames under the transcriptional control of the simian virus 40 early promoter. Transfection of Epstein-Barr virus-immortalized B-cell lines with this panel of recombinants consistently leads to stable expression of the HBV envelope, nucleocapsid, and polymerase proteins. The HBV X gene product is transiently expressed following transfection, but stable expression of this protein cannot be maintained on a long-term basis. To assess the suitability of this system for the identification of HBV-specific CTL in humans, a panel of EBO-HBV transfectants of defined HLA haplotype was used to monitor the HBV-specific CTL response in a patient with acute viral hepatitis type B. Transfectants that stably express the HBV nucleocapsid (core) antigen were found to serve as excellent targets for the detection of HLA class I-restricted CTL that recognize endogenously synthesized HBV core antigen in this patient; they were also successfully used to stimulate the specific expansion of these CTL in vitro.
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页码:2670 / 2678
页数:9
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