Unit

UNIT 7.10 Measurement of Proliferative Responses of Cultured Lymphocytes

  1. Linda Mesler Muul1,
  2. Guido Heine2,3,
  3. Christopher Silvin1,
  4. Stephen P. James4,
  5. Fabio Candotti1,
  6. Andreas Radbruch3,
  7. Margitta Worm3

Published Online: 1 AUG 2011

DOI: 10.1002/0471142735.im0710s94

Current Protocols in Immunology

Current Protocols in Immunology

How to Cite

Muul, L. M., Heine, G., Silvin, C., James, S. P., Candotti, F., Radbruch, A. and Worm, M. 2011. Measurement of Proliferative Responses of Cultured Lymphocytes. Current Protocols in Immunology. 94:II:7.10:7.10.1–7.10.26.

Author Information

  1. 1

    GMBB, National Human Genome Institute, Bethesda, Maryland

  2. 2

    Klinik für Dermatologie, Venerologie und Allergologie, Charité - Universitätsmedizin Berlin, Berlin, Germany

  3. 3

    Deutsches Rheuma-Forschungszentrum Berlin, Berlin, Germany

  4. 4

    University of Maryland School of Medicine, Baltimore, Maryland

Publication History

  1. Published Online: 1 AUG 2011
  2. Published Print: 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 induce proliferation in 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 protocol, which contains methods to determine cell proliferation by incorporation of [3H]thymidine into DNA by nonradioactive methods, e.g., reduction of tetrazolium salts (MTT or WST-1). These protocols provide an estimate of cell proliferation indirectly by measuring DNA synthesis, and cell metabolic activity in an entire cell population, but no data on individual cells is obtained. A protocol for CFSE labeling allows direct detection of single proliferating cells and facilitates the quantification of cell divisions by flow cytometry according to the respective CFSE-dilution, and following costaining with fluorescent labeled antibodies, the characterization of subpopulations in the cell culture. Curr. Protoc. Immunol. 94:7.10.1-7.10.26. © 2011 by John Wiley & Sons, Inc.

Keywords:

  • lymphocytes;
  • proliferation;
  • mitogens/antigens;
  • CFSE;
  • mixed lymphocyte reaction (MLR);
  • WST-1