Flow cytometric analysis of activation markers on stimulated T cells and their correlation with cell proliferation

Cytometry. 1997 Jan 1;27(1):71-6. doi: 10.1002/(sici)1097-0320(19970101)27:1<71::aid-cyto9>3.0.co;2-o.


The expression of activation antigens, namely CD25, CD69, CD71, and HLA-DR on T cells from 15 healthy individuals stimulated with different mitogens and specific antigens was evaluated by immunofluorescence assay and flow cytometric analysis and compared with cell proliferation as a function of [3H]thymidine incorporation. CD69 was the earliest expressed antigen on stimulated cells, while HLA-DR was the latest. Regardless of the stimulus used, lymphocytes expressing CD25 and CD71 were always more numerous than cells expressing CD69 and HLA-DR. Variations in the proportion of CD4+ and CD8+ T cells expressing each activation marker were observed with different antigenic stimuli. The expression of each activation marker showed overall agreement with the [3H]thymidine incorporation assay in discriminating between positive and negative immune response. However, no correlation was observed between the percentage of CD25-, CD69-, CD71-, and HLA-DR-positive T cells and the amount of [3H]thymidine incorporation. Moreover, low doses of mitogens and antigens as well as short time of stimulation were sufficient to induce T cells to express activation antigens but not to proliferate. Our data show that results obtained by flow cytometry and [3H]thymidine incorporation may differ qualitatively, at least under certain conditions; this suggests that the 2 assays are complementary, and when combined, may gives a clearer understanding of events leading to efficient cell-mediated immune response.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Antigens, CD / biosynthesis*
  • Cell Division
  • Cells, Cultured
  • DNA / biosynthesis
  • Flow Cytometry / methods*
  • HLA-DR Antigens / biosynthesis*
  • Humans
  • Leukocytes, Mononuclear / immunology
  • Lymphocyte Activation*
  • Lymphocyte Subsets
  • Mitogens / pharmacology
  • Molecular Sequence Data
  • T-Lymphocytes / immunology*
  • Thymidine / metabolism


  • Antigens, CD
  • HLA-DR Antigens
  • Mitogens
  • DNA
  • Thymidine