RAPID FLUOROMETRIC ASSAY OF HIV-INDUCED CELL FUSION
RAPID FLUOROMETRIC ASSAY OF HIV-INDUCED CELL FUSION
批准号:
3489609
负责人:
PAUL J MADDON
金额:
$5.0万
依托单位国家:
美国
项目类别:
财政年份:
1992
资助国家:
美国
项目状态:
已结题
起止时间:
1992-04-01 至 1993-01-31
关键词:
AIDS CD4 molecule CHO cells HIV envelope protein gp120 HIV envelope protein gp41 HeLa cells acidity /alkalinity antireceptor antibody antiviral antibody cell fusion clone cells fluorescent dye /probe fluorimetry gene expression helper T lymphocyte human immunodeficiency virus 1 immunologic assay /test membrane fusion method development mixed tissue /cell culture neoplastic cell culture for noncancer research pathologic process rapid diagnosis receptor expression temperature western blottings
中文摘要
表达HIV-1包膜糖蛋白gp120/gp41的细胞可以融合
英文摘要
Cells which express gp120/gp41, the HIV-1 envelope glycoprotein, can fuse
with cells expressing CD4, the HIV receptor, to form multi-nucleated giant
cells or syncytia. Such cell-to-cell fusion appears to be a mechanism for
virus transmission as well as a primary cause of cell death, and may play
an important role in the development of AIDS.
Current methods for measuring HIV-induced cell fusion are based on the
production of syncytia in CD4+ cells infected with HIV or in cells
expressing env using a transient expression system. Such assays typically
require several days, give variable results, and have a visual readout
which is both labor intensive and subjective. The first goal of this Phase
1 project is to develop a reproducible, virus-free cell fusion assay based
on syncytium formation between cells stably expressing HIV-1 gp120/gp41 and
CD4. The second goal is to demonstrate that fusion between these cells can
be assayed rapidly and with specificity using the fluorescence dequenching
(FDQ) method. Such an assay would be invaluable for detecting membrane
fusion in real time and for accurately measuring fusion kinetics.
In Phase 2, the FDQ fusion assay would be used in basic research studies of
HIV-induced cell fusion, including the identification of molecules, in
addition to CD4 and gp120/gp41, which are involved in membrane fusion. By
establishing new cell lines with env genes derived from different strains
of HIV-1, the assay would be useful in examining the fusogenic properties
of different isolates. Several applications of this assay would be
investigated, including the development of new clinical markers for HIV
disease, such as the presence of anti-syncytial antibodies and the
appearance of syncytium-inducing isolates. Finally, the assay would serve
as a model of virus-cell fusion, permitting the identification and
development of novel anti-fusion compounds and monoclonal neutralizing
antibodies which inhibit viral entry.
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资助金额:$54.6万
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财政年份:1998
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财政年份:1996
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财政年份:1996
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依托单位:
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资助金额:$81.34万
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财政年份:1996
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RAPID FLUOROMETRIC ASSAY OF HIV-INDUCED CELL FUSION
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批准号:2067703
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项目类别:
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资助金额:$19.64万
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财政年份:1992
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RAPID FLUOROMETRIC ASSAY OF HIV-INDUCED CELL FUSION
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批准号:2067704
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资助金额:$26.75万
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依托单位:
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批准号:2066683
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项目类别:
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财政年份:1991
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依托单位:
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财政年份:1991
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依托单位:
海外基金