Noninvasive immune monitoring assessed by flow cytometry and real time RT-PCR in urine of renal transplantation recipients
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2006-08-01
Autores
Galante, Nelson Zocoler
Saraiva Camara, Niels Olsen
Kallas, Esper Georges
Salomão, Reinaldo [UNIFESP]
Pacheco-Silva, Alvaro
Medina-Pestana, Jose Osmar
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Background: Monitoring recipient's alloreactivity has shown to be critical for limiting overimmunosuppression besides allowing preemptive treatment of acute rejection (AR).Methods: Flow cytometry and real time RT-PCR were performed in urine of kidney transplant recipients with AR (n=13) and compared with pyelonephritis(n = 10), chronic allograft nephropathy (n=13), acute tubular necrosis (n= 13) and stable graft function (n= 11). Expression of CD3, CD4, CD8, HLA-DR, Fas-L, ICAM-1 and CD25 were assessed using flow cytometry. mRNA of perform, granzyme B and Fas-L were quantified by real time RT-PCR.Results: Frequencies of CD3(+), HLA-DR+, Fas-L+, ICAM-1(+) and CD25(+) cells were significantly higher in AR group (P<0.05). ROC curves showed sensitivity from 70% to 91% and specificity from 30% to 100%, whereas the highest sensitivity and specificity was 91% and 100% respectively, for Fas-L+ cells. Levels of mRNA of perforin, granzyme B and Fas-L were significantly augmented in AR, while the sensitivity and specificity ranged from 85% to 88% and from 55% to 100%, respectively.Conclusions: Analyses of immune activation markers by flow cytometry and real time RT-PCR are equally useful for noninvasive monitoring kidney allografts. (C) 2006 Elsevier B.V. All rights reserved.
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Transplant Immunology. Amsterdam: Elsevier B.V., v. 16, n. 2, p. 73-80, 2006.