Unit

UNIT 7.10 Measurement of Proliferative Responses of Cultured Lymphocytes

  1. Linda Mesler Muul1,
  2. Christopher Silvin1,
  3. Stephen P. James2,
  4. Fabio Candotti1

Published Online: 1 AUG 2008

DOI: 10.1002/0471142735.im0710s82

Current Protocols in Immunology

Current Protocols in Immunology

How to Cite

Muul, L. M., Silvin, C., James, S. P. and Candotti, F. 2008. Measurement of Proliferative Responses of Cultured Lymphocytes. Current Protocols in Immunology. 82:II:7.10:7.10.1–7.10.24.

Author Information

  1. 1

    GMBB, National Human Genome Institute, Bethesda, Maryland

  2. 2

    University of Maryland School of Medicine, Baltimore, Maryland

Publication History

  1. Published Online: 1 AUG 2008
  2. Published Print: AUG 2008

This is not the most recent version of the article. View current version (1 AUG 2011)

Abstract

Measurement of proliferative responses of human lymphocytes is a fundamental technique for the assessment of their biological responses to various stimuli. Most simply, this involves measurement of the number of cells present in a culture before and after the addition of a stimulating agent. This unit contains several different prototype protocols to measure the proliferative response of lymphocytes following exposure to mitogens, antigens, allogeneic or autologous cells, or soluble factors. Each of these protocols can be used in conjunction with an accompanying support protocol which contains methods for pulsing cultures with [3H]thymidine and determining incorporation of [3H]thymidine into DNA or assessing cell proliferation by nonradioactive methods, e.g., reduction of tetrazolium salts (MTT). The protocols described here provide an estimate of DNA synthesis and cell proliferation in an entire cell population, but do not provide information on the proliferation of individual cells. A protocol for CFSE labeling allows specific subpopulations of cells to be separated viably for further analysis. Curr. Protoc. Immunol. 82:7.10.1-7.10.24. © 2008 by John Wiley & Sons, Inc.

Keywords:

  • lymphocytes;
  • proliferation;
  • mitogens/antigens;
  • CFSE;
  • mixed lymphocyte reaction (MLR)