Mr. Michael R. Warnement Profile

Michael R. Warnement

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Proceedings Article | 27 March 2006 Paper

Interrogating the signaling dynamics of T cell activation with quantum dots

Michael Warnement, Shannon Faley, John Wikswo, Sandra Rosenthal

Proceedings Volume 6096, 60960T (2006) https://doi.org/10.1117/12.663345

KEYWORDS: Receptors, Signal detection, Quantum dots, Luminescence, Confocal microscopy, Pathogens, Multiplexing, Biological detection systems, Calcium, Biological weapons

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We report the use of antibody-conjugated quantum dots (QDs) to monitor the expression dynamics of the membrane bound cytokine receptor interleukin-2 receptor-α (IL-2Rα) throughout the course of Jurkat T cell activation. Maximal receptor expression is observed 32-48 hours after activation, followed by a sharp decrease subsequent to 48 hours consistent with IL-2R internalization. Fluorescence microscopy, ELISA, and FACS analyses were used to verify controlled activation and specificity of QD labeling. Additionally, confocal microscopy demonstrated receptor internalization subsequent to expression and QD labeling. Antibody-conjugated QDs provide a convenient means to rapidly determine cell state and interrogate end products of cell signaling pathways. Interrogation of other signaling pathways can eventually be carried out in a similar manner upon identification of relevant membrane associated receptors. Ultimately, the multiplexing capabilities of QDs will allow the examination of several signaling pathways simultaneously and aid in toxin detection and discrimination.

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