TY - JOUR
T1 - A quantitative high-throughput chemotaxis assay using bioluminescent reporter cells
AU - Vishwanath, Reena P.
AU - Brown, Christine E.
AU - Wagner, Jamie R.
AU - Meechoovet, Hunsar B.
AU - Naranjo, Araceli
AU - Wright, Christine L.
AU - Olivares, Simon
AU - Qian, Dajun
AU - Cooper, Laurence J.N.
AU - Jensen, Michael C.
N1 - Copyright:
Copyright 2008 Elsevier B.V., All rights reserved.
PY - 2005/7
Y1 - 2005/7
N2 - Here we report on a novel biophotonic assay system for the detection and quantitation of chemotaxis, the directed movement of cells in response to chemokine concentration gradients. Our assay employs a firefly luciferase (ffLuc)-generated biophotonic signal to quantify cellular migration in 96-well microplate chemotaxis instruments. When compared to direct cell enumeration, the biophotonic reporter method is superior in accuracy, reproducibility, and sensitivity. As a proof-of-concept, we demonstrate the utility of this assay for quantifying the chemotactic response of ex vivo expanded ffLuc+ primary human T-cells to recombinant human chemokines MCP-1, RANTES, and IP-10. The 96-well microplate format and in situ biophotonic detection of cells are amenable to high-throughput screening of peptides and small molecule libraries to identify agonists and antagonists of cellular chemotaxis, to analyze biological fluids for chemotactic activity, and to study chemotaxis in a variety of cell types.
AB - Here we report on a novel biophotonic assay system for the detection and quantitation of chemotaxis, the directed movement of cells in response to chemokine concentration gradients. Our assay employs a firefly luciferase (ffLuc)-generated biophotonic signal to quantify cellular migration in 96-well microplate chemotaxis instruments. When compared to direct cell enumeration, the biophotonic reporter method is superior in accuracy, reproducibility, and sensitivity. As a proof-of-concept, we demonstrate the utility of this assay for quantifying the chemotactic response of ex vivo expanded ffLuc+ primary human T-cells to recombinant human chemokines MCP-1, RANTES, and IP-10. The 96-well microplate format and in situ biophotonic detection of cells are amenable to high-throughput screening of peptides and small molecule libraries to identify agonists and antagonists of cellular chemotaxis, to analyze biological fluids for chemotactic activity, and to study chemotaxis in a variety of cell types.
KW - Bioluminescence
KW - Chemotaxis
KW - Luciferase
KW - Lymphocyte
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U2 - 10.1016/j.jim.2005.04.021
DO - 10.1016/j.jim.2005.04.021
M3 - Article
C2 - 15987642
AN - SCOPUS:23444457625
SN - 0022-1759
VL - 302
SP - 78
EP - 89
JO - Journal of Immunological Methods
JF - Journal of Immunological Methods
IS - 1-2
ER -