Stimulation of murine T cell subsets with anti-CD3 antibody. Age-related defects in the expression of early activation molecules

J Immunol. 1989 Mar 1;142(5):1413-21.


Splenocytes from young (3 to 4 mo) and aged (24 to 26 mo) C57BL/6 mice were stimulated with anti-CD3 epsilon mAb in vitro. At the time of peak DNA synthesis (day 2), cells from aged mice incorporated congruent to 60% less [3H]TdR than cells from young mice. This age-related defect was not attributable to gross differences in anti-CD3 does optima, response kinetics, accessory cell function, numbers of T cells cultured, CD4+:CD8+ cell ratios or surface levels of CD3 epsilon molecules. In an attempt to analyze pre-S phase events in these responses, we monitored CD4+ and CD8+ cells in splenocyte cultures for the time-dependent expression of three T cell activation markers: RL388 Ag and IL-2R and transferrin R. Parallel analyses of mean T cell size and cell cycle phase distributions were performed. Non-activated T cells from both age groups similarly expressed moderate levels of RL388 Ag, low levels of transferrin R, and undetectable levels of IL-2R. Analysis of stimulated T cells revealed, in both age groups: 1) detectable increases in expression of all three markers by 6 h of culture, and continued increases associated with blastogenesis and G1 phase transit and 2) a preferential stimulation of the CD8+ subset to a state of high level marker expression. Age group comparisons of activation marker expression over time suggested that the age-related defect reflects proportionally smaller fractions of CD4+ and CD8+ cells that respond normally, rather than a general defect in all T cells or a subset-specific defect. Finally, we found that supernatants from aged donor cell cultures stimulated with anti-CD3 contained less Il-2 than those of young controls. Addition of an IL-2 containing supernatant to aged donor cell cultures increased, but did not restore, the S phase response on day 2; however, the response on day 3 was comparable to the peak (day 2) response of young controls. These data suggest that exogenous IL-2 can improve the aged response, perhaps by expanding the fraction of normally reactive T cells.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Aging*
  • Animals
  • Antibodies, Monoclonal / physiology*
  • Antigens, CD*
  • Antigens, Differentiation, T-Lymphocyte / analysis*
  • Antigens, Differentiation, T-Lymphocyte / immunology*
  • CD3 Complex
  • DNA / biosynthesis
  • Female
  • Interleukin-2 / biosynthesis
  • Lectins, C-Type
  • Lymphocyte Activation*
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Phenotype
  • Rats
  • Rats, Inbred Lew
  • Receptors, Antigen, T-Cell / immunology*
  • Spleen
  • T-Lymphocytes / classification
  • T-Lymphocytes / immunology*
  • T-Lymphocytes / metabolism


  • Antibodies, Monoclonal
  • Antigens, CD
  • Antigens, Differentiation, T-Lymphocyte
  • CD3 Complex
  • CD69 antigen
  • Interleukin-2
  • Lectins, C-Type
  • Receptors, Antigen, T-Cell
  • DNA