Non-invasive diagnosis of acute rejection in kidney transplants with delayed graft function.

Delayed graft function (DGF) often occurs in kidney transplants from deceased donors. We wanted to provide studies giving more accurate non-invasive tests for acute rejection (AR). Using real-time PCR, we examined the expression of cytolytic molecules such as perforin, granzyme B, and fas-ligand along with serpin proteinase inhibitor-9. We also measured the expression of FOXP3, a characteristic gene of T-regulatory cells known to be involved in AR. These studies were conducted on peripheral blood monocytes, urinary cells, and 48 surveillance kidney biopsies taken from a total of 35 patients with DGF. Of these patients, 20 had a histopathological diagnosis of AR, whereas other 28 had characteristics of acute tubular necrosis (ATN). Expression of cytolytic and apoptotic-associated genes in the biopsy tissue, peripheral blood leukocytes, and urinary cells was significantly higher in patients with AR than that in patients with ATN. Diagnostic parameters associated with FOXP3 gene expression were most accurate in peripheral blood leukocytes and urine cells with sensitivity, specificity, positive and negative predictive values, and accuracy between 94 and 100%. Our study shows that quantification of selected genes in peripheral blood leukocytes and urinary cells from renal transplant patients with DGF may provide a useful and accurate non-invasive diagnosis of AR.

Evaluation of apoptosis in peripheral blood lymphocytes of renal transplant patients.

The aim of this study was to evaluate the occurrence of apoptosis and the expression of FasL and IL-2 genes in apoptotic peripheral blood mononuclear cells (PBMC) at different posttransplant periods. Three groups of patients were studied: group 1, kidney transplant recipients at least 1 year posttransplant (n = 17); group 2, kidney transplant recipients at least 5 years posttransplant (n = 15); and a control group composed of 7 healthy subjects. Apoptosis was detected by annexin flow cytometry and gene expression by reverse transcription polymerase chain reactions. The percentage of apoptotic cells was significantly higher in groups 1 (42 +/- 4%) and 2 (37 +/- 3%) than the controls (27 +/- 2%; P < .0001). Apoptotic cells in group 1 was significantly higher than in group 2 (P < .005). A significant difference in FasL expression was observed between groups 1 and 2 (P < .001) and the immunosuppressive regimen. These findings suggest that PBMC of kidney transplant recipients are more susceptible to activation-induced cell death and that the Fas-FasL pathway is involved in this process.