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High Throughput Method to Quantify Anterior-Posterior Polarity of T-Cells and Epithelial Cells
Department of Molecular Virology and Microbiology, Baylor College of Medicine, One Baylor Plaza, Houston, TX 77030, USA
Department of Molecular and Cellular Biology, Baylor College of Medicine, One Baylor Plaza, Houston, TX 77030, USA
* Author to whom correspondence should be addressed.
Received: 23 September 2011; in revised form: 16 November 2011 / Accepted: 23 November 2011 / Published: 28 November 2011
Abstract: The virologic synapse (VS), which is formed between a virus-infected and uninfected cell, plays a central role in the transmission of certain viruses, such as HIV and HTLV-1. During VS formation, HTLV-1-infected T-cells polarize cellular and viral proteins toward the uninfected T-cell. This polarization resembles anterior-posterior cell polarity induced by immunological synapse (IS) formation, which is more extensively characterized than VS formation and occurs when a T-cell interacts with an antigen-presenting cell. One measure of cell polarity induced by both IS or VS formation is the repositioning of the microtubule organizing center (MTOC) relative to the contact point with the interacting cell. Here we describe an automated, high throughput system to score repositioning of the MTOC and thereby cell polarity establishment. The method rapidly and accurately calculates the angle between the MTOC and the IS for thousands of cells. We also show that the system can be adapted to score anterior-posterior polarity establishment of epithelial cells. This general approach represents a significant advancement over manual cell polarity scoring, which is subject to experimenter bias and requires more time and effort to evaluate large numbers of cells.
Keywords: polarity; high throughput imaging; immunological synapse; virological synapse; T-cell; epithelial cell; pipeline pilot
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Irvin-Wilson, C.V.; Newberg, J.Y.; Kong, K.; Javier, R.T.; Marriott, S.J. High Throughput Method to Quantify Anterior-Posterior Polarity of T-Cells and Epithelial Cells. Viruses 2011, 3, 2396-2411.
Irvin-Wilson CV, Newberg JY, Kong K, Javier RT, Marriott SJ. High Throughput Method to Quantify Anterior-Posterior Polarity of T-Cells and Epithelial Cells. Viruses. 2011; 3(12):2396-2411.
Irvin-Wilson, Charletha V.; Newberg, Justin Y.; Kong, Kathleen; Javier, Ronald T.; Marriott, Susan J. 2011. "High Throughput Method to Quantify Anterior-Posterior Polarity of T-Cells and Epithelial Cells." Viruses 3, no. 12: 2396-2411.