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A quantitative high-throughput chemotaxis assay using bioluminescent reporter cells.
Vishwanath, Reena P; Brown, Christine E; Wagner, Jamie R; Meechoovet, Hunsar B; Naranjo, Araceli; Wright, Christine L; Olivares, Simon; Qian, Dajun; Cooper, Laurence J N; Jensen, Michael C.
Afiliação
  • Vishwanath RP; Division of Cancer Immunotherapeutics and Tumor Immunology, Beckman Research Institute, City of Hope National Medical Center, Duarte, CA 91010, USA.
J Immunol Methods ; 302(1-2): 78-89, 2005 Jul.
Article em En | MEDLINE | ID: mdl-15987642
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.
Assuntos
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Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Linfócitos T / Quimiotaxia de Leucócito / Genes Reporter / Luciferases Limite: Animals / Humans Idioma: En Revista: J Immunol Methods Ano de publicação: 2005 Tipo de documento: Article País de afiliação: Estados Unidos
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Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Linfócitos T / Quimiotaxia de Leucócito / Genes Reporter / Luciferases Limite: Animals / Humans Idioma: En Revista: J Immunol Methods Ano de publicação: 2005 Tipo de documento: Article País de afiliação: Estados Unidos