Unit

UNIT 6.28 Measurement of T Cell Activation After 16-hr In Vitro Stimulation with Concanavalin A

  1. Borros M. Arneth

Published Online: 1 JAN 2010

DOI: 10.1002/0471142956.cy0628s51

Current Protocols in Cytometry

Current Protocols in Cytometry

How to Cite

Arneth, B. M. 2010. Measurement of T Cell Activation After 16-hr In Vitro Stimulation with Concanavalin A. Current Protocols in Cytometry. 51:6.28:6.28.1–6.28.10.

Author Information

  1. Institute of Clinical Chemistry and Laboratory Medicine, Johannes Gutenberg University Mainz, Mainz, Germany

Publication History

  1. Published Online: 1 JAN 2010
  2. Published Print: JAN 2010

Abstract

A flow cytometry assay that can be used to directly determine the proportion of activated T lymphocytes in human whole blood samples after stimulation with concanavalin A is presented here. Human whole blood is incubated with fluorescently labeled antibodies (against CD3, CD4, CD8, and CD69), erythrocytes are then lysed, and the samples are analyzed using a flow cytometer. The assay presented is able to differentiate between CD4+ and CD8+ T lymphocytes. Thus, it is possible to quantify both lymphocyte populations in parallel, as well as the respective proportions of activated T lymphocytes, all from one sample. An additional advantage of this assay is that it was developed to assay whole blood, thereby reducing the likelihood of introducing pre-analytic error and facilitating the calculation of quantitative ratios. Thus, the method can be used for both in vitro and ex vivo experiments. Curr. Protoc. Cytom. 51:6.28.1-6.28.10. © 2010 by John Wiley & Sons, Inc.

Keywords:

  • activated T cells;
  • lymphocytes;
  • lymphocyte proliferation assay;
  • activation marker;
  • CD4 T helper cells;
  • CD8 cytotoxic T cells;
  • CD69 activation marker;
  • T lymphocytes;
  • CD69;
  • CD4 T cells