Evidence that increases in lymphocyte tyrosine phosphorylation precede cardiac allograft rejection. Effects of cyclosporine and potential use in clinical management.
Tyrosine phosphorylation is an early, critical event in lymphocyte signal transduction. We measured tyrosine phosphorylation in a porcine experimental transplant model to evaluate its utility in monitoring the allograft immune response. Using flow cytometry, we demonstrate a biphasic increase in phosphotyrosine (ptyr) levels in peripheral blood mononuclear cells (PBMC), and that increases are detectable as early as 1 day posttransplantation in untreated transplanted animals (n = 4). This biphasic response is likely result from the sequestration of ptyr+ cells from the periphery into the graft as graft-infiltrating lymphocytic cells show increased ptyr levels. This suggests possible lymphocyte trafficking between the peripheral compartment and the allograft. A 5-day course of treatment with cyclosporine (CsA) at 20 mg/kg/day (n = 4), but not at 10 mg/kg/day (n = 4), prevents graft rejection in this allograft model. Strikingly, treatment with 20 mg/kg/day CsA, but not with 10 mg/kg/day, suppressed increases in ptyr levels in both PBMC and graft-infiltrating cells. Increases in ptyr levels in PBMC are detectable 2-5 days before histologic and electrocardiographic signs of graft rejection, suggesting a potential diagnostic utility for measuring tyrosine phosphorylation in monitoring and managing transplant rejection.
Tsao
P W
PW
Department of Pharmacology, University of Toronto, Canada.
Mills
G B
GB
Diaz
R J
RJ
Radde
I C
IC
Parkinson
D
D
Waddell
J
J
Wilson
G J
GJ
Coles
J G
JG
D000818:Animals; D015415:Biomarkers; D016572:Cyclosporine / Q000627:therapeutic use*; D005434:Flow Cytometry; D006084:Graft Rejection / Q000175:diagnosis* / Q000517:prevention & control; D016027:Heart Transplantation; D007124:Immunoenzyme Techniques; D016130:Immunophenotyping; D008214:Lymphocytes / Q000378:metabolism*; D009206:Myocardium / Q000378:metabolism; D010766:Phosphorylation; D019000:Phosphotyrosine; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D015398:Signal Transduction; D013552:Swine; D014184:Transplantation, Homologous; D014443:Tyrosine / Q000031:analogs & derivatives / Q000032:analysis / Q000378:metabolism*
D015415:Biomarkers; D019000:Phosphotyrosine; D014443:Tyrosine; D016572:Cyclosporine