Detection of cardiac allograft rejection by real-time PCR analysis of circulating mononuclear cells.
BACKGROUND
Detection of cardiac allograft rejection is based on the histological examination of endomyocardial biopsies (EMB). We have explored the possibility of whether graft rejection could be detected by characteristic gene expression patterns in peripheral blood mononuclear cells (PBMC) of heart-transplant recipients.


METHODS
The study included 58 blood samples of 44 patients. On the day of EMB, mononuclear cells were isolated from peripheral blood, and gene expression was measured by quantitative real-time PCR. Thirty-nine parameters, including cytokine and chemokine genes were analyzed. Gene expression results were correlated with histological assessment of concomitant evaluated EMB according to International Society for Heart and Lung Transplantation (ISHLT) nomenclature.


RESULTS
Gene expression of perforin, CD95 ligand, granzyme B, RANTES, CXCR3, COX2, ENA 78 and TGF-beta1 was significantly different in PBMC of patients with mild to moderate degrees of allograft rejection (> or =grade 2) compared with patients exhibiting no or minor forms of rejection (<grade 2). Using discriminance analysis, five parameters were found that allow discrimination of rejection > or =grade 2 vs. <grade 2 with a sensitivity of 84% and a specificity of 82% as assessed by receiver operating characteristic analysis.


CONCLUSIONS
Quantitative analysis of gene expression in PBMC may be a valuable tool for non-invasive diagnosis of allograft rejection and may allow further insight in the biological process of graft rejection.
Schoels
M
M
Institute of Immunology, University of Heidelberg, Heildelberg, Germany.
Dengler
T J
TJ
Richter
R
R
Meuer
S C
SC
Giese
T
T
D000954:Antigens, Surface / Q000235:genetics; D017209:Apoptosis / Q000235:genetics; D018946:Chemokine CCL5 / Q000235:genetics; D054365:Chemokine CXCL5; D019743:Chemokines, CXC / Q000235:genetics; D002634:Chemotaxis, Leukocyte / Q000235:genetics; D051546:Cyclooxygenase 2; D053222:Fas Ligand Protein; D005260:Female; D005786:Gene Expression Regulation / Q000235:genetics; D006084:Graft Rejection / Q000175:diagnosis* / Q000473:pathology; D053804:Granzymes; D016027:Heart Transplantation; D006801:Humans; D007527:Isoenzymes / Q000235:genetics; D007963:Leukocytes, Mononuclear / Q000378:metabolism*; D008024:Ligands; D008297:Male; D008562:Membrane Glycoproteins / Q000235:genetics; D008565:Membrane Proteins; D008875:Middle Aged; D054353:Perforin; D016133:Polymerase Chain Reaction; D052899:Pore Forming Cytotoxic Proteins; D011451:Prostaglandin-Endoperoxide Synthases / Q000235:genetics; D054367:Receptors, CXCR3; D019707:Receptors, Chemokine / Q000235:genetics; D012680:Sensitivity and Specificity; D012697:Serine Endopeptidases / Q000235:genetics; D016212:Transforming Growth Factor beta / Q000235:genetics; D053773:Transforming Growth Factor beta1; D019014:fas Receptor / Q000235:genetics
D000954:Antigens, Surface; C486263:CXCL5 protein, human; C515187:CXCR3 protein, human; D018946:Chemokine CCL5; D054365:Chemokine CXCL5; D019743:Chemokines, CXC; C505661:FASLG protein, human; D053222:Fas Ligand Protein; D007527:Isoenzymes; D008024:Ligands; D008562:Membrane Glycoproteins; D008565:Membrane Proteins; D052899:Pore Forming Cytotoxic Proteins; D054367:Receptors, CXCR3; D019707:Receptors, Chemokine; C508960:TGFB1 protein, human; D016212:Transforming Growth Factor beta; D053773:Transforming Growth Factor beta1; D019014:fas Receptor; D054353:Perforin; D051546:Cyclooxygenase 2; C496540:PTGS2 protein, human; D011451:Prostaglandin-Endoperoxide Synthases; C509213:GZMB protein, human; D053804:Granzymes; D012697:Serine Endopeptidases