Novel apoptosis assay to analyze immunosuppression

Cytometry A. 2006 Mar;69(3):158-60. doi: 10.1002/cyto.a.20218.

Abstract

Background: We developed a novel whole blood assay to assess the effects of immunosuppressants on lymphocyte apoptosis.

Methods: Peripheral blood for six experiments for each drug was treated either with cyclosporin A (CsA), mycophenolate acid (MPA), tacrolimus (TRL) or rapamycin (RAPA). Whole blood was stimulated with different concentrations of staurosporine (0-5 microM) for 24 h. Using FACS, apoptosis were measured by Annexin V expression (%cells +/- SEM).

Results: MPA, RAPA, TRL, but not CsA, increased (P < 0.05) apoptotic cells (MPA:20.9 +/- 3.7; RAPA:17.5 +/- 3.3; TRL:16.4 +/- 2.9 vs. control:15.2 +/- 2.8).

Conclusion: This new whole blood assay exhibited that MPA has a stronger effect on apoptosis than RAPA and TRL, whereas CsA had no effect.

MeSH terms

  • Annexin A5 / analysis
  • Apoptosis / drug effects*
  • Cyclosporine / pharmacology
  • Flow Cytometry
  • Humans
  • Immunosuppression Therapy*
  • Immunosuppressive Agents / pharmacology*
  • In Situ Nick-End Labeling
  • Lymphocytes / drug effects*
  • Mycophenolic Acid / pharmacology
  • Sirolimus / pharmacology
  • Staurosporine / pharmacology
  • Tacrolimus / pharmacology

Substances

  • Annexin A5
  • Immunosuppressive Agents
  • Cyclosporine
  • Staurosporine
  • Mycophenolic Acid
  • Sirolimus
  • Tacrolimus