Journal article
A novel quantum dot-based high throughput fluorescence assay for the quantitation of human T cell leukemia virus type 1 binding to host cells
Journal of neurovirology, Vol.12, pp.37-37
01 May 2006
Abstract
Quantum dots (QD) are fluorescent semiconductor nanocrystals comprised of a cadmium selenide core with a semiconductor shell of zinc sulfide coated with a polymer shell allowing particles to be conjugated to biological molecules while retaining the optical properties of the particle. They have narrow emission spectrums, high quantum yield, and good photostability. We have used these unique properties of QDs to develop a simple high-throughput binding assay to study the attachment of HTLV-1 to host cells. In order to enhance the end point detection limit, we have biotinylated HTLV-1 (biot-HTLV-1) and used streptavidin-coated QDs (strep-QD). Primary standardization was performed on Jurkat CD4+ T cells since this cell type is representative of the major target population infected with HTLV-1 in patients. A dose-dependent increase was observed in HTLV-1 binding to Jurkat cells indicating linearity of the assay at both 4 degree and 37 degree C. The specificity of the assay was confirmed by using increasing amounts of non-biotinylated virus to demonstrate competitive inhibition. We have also tested differential binding of HTLV-1 to various cell types of pathogenic relevance. T cells and dendritic cells (DCs) exhibited maximal affinity as compared to other cell types examined. Finally, we have screened a number of blocking antibodies directed against a putative HTLV-1 receptor on DCs referred to as DC-SIGN (dendritic cell specific ICAM-3 grabbing non-integrin). We were able to map DC-SIGN epitopes important for HTLV-1 binding as compared to those for HIV-1 binding. This novel, easy-to-perform, high throughput assay can be utilized to study the binding of any virus that can be biotinylated, for the small molecule inhibitor screening, epitope mapping, and for receptor studies.
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Details
- Title
- A novel quantum dot-based high throughput fluorescence assay for the quantitation of human T cell leukemia virus type 1 binding to host cells
- Creators
- K KampaniJ AhujaB WigdahlP Jain
- Publication Details
- Journal of neurovirology, Vol.12, pp.37-37
- Resource Type
- Journal article
- Language
- English
- Academic Unit
- Microbiology and Immunology
- Identifiers
- 991019170573804721