Application of HIV-1-green fluorescent protein (GFP) reporter viruses in neutralizing antibody assays
We made reporter HIV-1 DNA constructs carrying green fluorescent protein (GFP) gene and exchangeable env of subtype E. The recombinant constructs were used to produce infectious reporter viruses, which induced infected cells to emit green fluorescent light and rendered them easily detectable at sing...
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th-mahidol.265532018-09-07T16:45:25Z Application of HIV-1-green fluorescent protein (GFP) reporter viruses in neutralizing antibody assays P. Auewarakul V. Paungcharoen S. Louisirirotchanakul C. Wasi Mahidol University Immunology and Microbiology Medicine We made reporter HIV-1 DNA constructs carrying green fluorescent protein (GFP) gene and exchangeable env of subtype E. The recombinant constructs were used to produce infectious reporter viruses, which induced infected cells to emit green fluorescent light and rendered them easily detectable at single cell level. Because the env in this construct can be easily exchanged, viruses with different antigenic epitopes can be made. We used these reporter viruses to set up a neutralizing antibody assay based on fluorescence reduction by flow cytometric measurement. The result of this new assay correlated with the standard infectivity reduction assay using primary isolates. Because this new assay is faster and much less costly than the standard assay using a p24 endpoint and can be performed in peripheral blood mononuclear cells (PBMC), it provides a useful tool for analysis of HIV-1 immune responses. 2018-09-07T09:41:07Z 2018-09-07T09:41:07Z 2001-11-17 Article Asian Pacific Journal of Allergy and Immunology. Vol.19, No.2 (2001), 139-144 0125877X 2-s2.0-0034767704 https://repository.li.mahidol.ac.th/handle/123456789/26553 Mahidol University SCOPUS https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=0034767704&origin=inward |
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Immunology and Microbiology Medicine P. Auewarakul V. Paungcharoen S. Louisirirotchanakul C. Wasi Application of HIV-1-green fluorescent protein (GFP) reporter viruses in neutralizing antibody assays |
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We made reporter HIV-1 DNA constructs carrying green fluorescent protein (GFP) gene and exchangeable env of subtype E. The recombinant constructs were used to produce infectious reporter viruses, which induced infected cells to emit green fluorescent light and rendered them easily detectable at single cell level. Because the env in this construct can be easily exchanged, viruses with different antigenic epitopes can be made. We used these reporter viruses to set up a neutralizing antibody assay based on fluorescence reduction by flow cytometric measurement. The result of this new assay correlated with the standard infectivity reduction assay using primary isolates. Because this new assay is faster and much less costly than the standard assay using a p24 endpoint and can be performed in peripheral blood mononuclear cells (PBMC), it provides a useful tool for analysis of HIV-1 immune responses. |
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Mahidol University |
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Mahidol University P. Auewarakul V. Paungcharoen S. Louisirirotchanakul C. Wasi |
format |
Article |
author |
P. Auewarakul V. Paungcharoen S. Louisirirotchanakul C. Wasi |
author_sort |
P. Auewarakul |
title |
Application of HIV-1-green fluorescent protein (GFP) reporter viruses in neutralizing antibody assays |
title_short |
Application of HIV-1-green fluorescent protein (GFP) reporter viruses in neutralizing antibody assays |
title_full |
Application of HIV-1-green fluorescent protein (GFP) reporter viruses in neutralizing antibody assays |
title_fullStr |
Application of HIV-1-green fluorescent protein (GFP) reporter viruses in neutralizing antibody assays |
title_full_unstemmed |
Application of HIV-1-green fluorescent protein (GFP) reporter viruses in neutralizing antibody assays |
title_sort |
application of hiv-1-green fluorescent protein (gfp) reporter viruses in neutralizing antibody assays |
publishDate |
2018 |
url |
https://repository.li.mahidol.ac.th/handle/123456789/26553 |
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1763494638850670592 |