title: Noninvasive immune monitoring assessed by flow cytometry and real time RT-PCR in urine of renal transplantation recipients.
abstract: 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 perforin, 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.
authors.affiliation: Laboratório de Imunologia Clínica e Experimental. Departamento de Medicina, Disciplina de Nefrologia, Universidade Federal de São Paulo-UNIFESP, São Paulo, Brazil.
authors.forename: Nelson Zocoler
authors.initials: NZ
authors.lastname: Galante
authors.forename: Niels Olsen Saraiva
authors.initials: NO
authors.lastname: Câmara
authors.forename: Esper Georges
authors.initials: EG
authors.lastname: Kallas
authors.forename: Reinaldo
authors.initials: R
authors.lastname: Salomão
authors.forename: Alvaro
authors.initials: A
authors.lastname: Pacheco-Silva
authors.forename: José Osmar
authors.initials: JO
authors.lastname: Medina-Pestana
mesh_terms: D005260:Female; D005434:Flow Cytometry; D006084:Graft Rejection / Q000276:immunology / Q000652:urine*; D053804:Granzymes; D006801:Humans; D007674:Kidney Diseases / Q000276:immunology / Q000652:urine; D016030:Kidney Transplantation / Q000009:adverse effects / Q000276:immunology; D008297:Male; D008562:Membrane Glycoproteins / Q000276:immunology / Q000652:urine; D008991:Monitoring, Physiologic; D054353:Perforin; D052899:Pore Forming Cytotoxic Proteins; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012697:Serine Endopeptidases / Q000276:immunology / Q000652:urine
chemical_list: D008562:Membrane Glycoproteins; D052899:Pore Forming Cytotoxic Proteins; D054353:Perforin; C509213:GZMB protein, human; D053804:Granzymes; D012697:Serine Endopeptidases