Unit

UNIT 4.7 Measurement of T and B Cell Turnover with Bromodeoxyuridine

  1. David F. Tough,
  2. Jonathan Sprent

Published Online: 1 MAY 2001

DOI: 10.1002/0471142735.im0407s18

Current Protocols in Immunology

Current Protocols in Immunology

How to Cite

Tough, D. F. and Sprent, J. 2001. Measurement of T and B Cell Turnover with Bromodeoxyuridine. Current Protocols in Immunology. 4:4.7.

Author Information

  1. The Scripps Research Institute, La Jolla, California

Publication History

  1. Published Online: 1 MAY 2001
  2. Published Print: JUN 1996

This is not the most recent version of the article. View current version (1 MAY 2007)

Abstract

The rate of turnover of T and B cells can be measured in terms of the time between two mitoses or between mitosis and death. In vivo, this is accomplished by administering DNA precursors and measuring the rate at which cells become labeled with these precursors. 5-bromo-2'-deoxyuridine (BrdU) is a thymidine analog that is incorporated into the DNA of cells during the S phase of the cell cycle and can be subsequently detected with a specific monoclonal antibody. In this method, BrdU is given to experimental animals either continuously or as a pulse, depending on the goal of the experiment. Subsequently, cells are stained in vitro with an anti-BrdU antibody and labeled cells detected by flow cytometry. By combining cell surface staining for multiple lymphocyte markers with anti-BrdU staining, it is possible to determine the lifespan of phenotypically defined subsets.