Novel apoptosis assay to analyze immunosuppression.
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).


CONCLUSIONS
This new whole blood assay exhibited that MPA has a stronger effect on apoptosis than RAPA and TRL, whereas CsA had no effect.
Department of Cardiac Surgery, University of Leipzig-Heart Center, Leipzig, Germany. andreasboldt@web.de
Andreas
A
Boldt
Markus Johannes
MJ
Barten
Claudia
C
Weiss
Anja
A
Sagner
Friedrich Wilhelm
FW
Mohr
Jan Fritz
JF
Gummert
D017304:Annexin A5 / Q000032:analysis; D017209:Apoptosis / Q000187:drug effects*; D016572:Cyclosporine / Q000494:pharmacology; D005434:Flow Cytometry; D006801:Humans; D007165:Immunosuppression Therapy; D007166:Immunosuppressive Agents / Q000494:pharmacology*; D020287:In Situ Nick-End Labeling; D008214:Lymphocytes / Q000187:drug effects*; D009173:Mycophenolic Acid / Q000494:pharmacology; D020123:Sirolimus / Q000494:pharmacology; D019311:Staurosporine / Q000494:pharmacology; D016559:Tacrolimus / Q000494:pharmacology
D017304:Annexin A5; D007166:Immunosuppressive Agents; D016572:Cyclosporine; D019311:Staurosporine; D009173:Mycophenolic Acid; D020123:Sirolimus; D016559:Tacrolimus