Messenger RNA for FOXP3 in the urine of renal-allograft recipients.
BACKGROUND
The outcome of renal transplantation after an episode of acute rejection is difficult to predict, even with an allograft biopsy.


METHODS
We studied urine specimens from 36 subjects with acute rejection, 18 subjects with chronic allograft nephropathy, and 29 subjects with normal biopsy results. Levels of messenger RNA (mRNA) for FOXP3, a specification and functional factor for regulatory T lymphocytes, and mRNA for CD25, CD3epsilon, perforin, and 18S ribosomal RNA (rRNA) were measured with a kinetic, quantitative polymerase-chain-reaction assay. We examined associations of mRNA levels with acute rejection, rejection reversal, and graft failure.


RESULTS
The log-transformed mean (+/-SE) ratio of FOXP3 mRNA copies to 18S ribosomal RNA copies was higher in urine from the group with acute rejection (3.8+/-0.5) than in the group with chronic allograft nephropathy (1.3+/-0.7) or the group with normal biopsy results (1.6+/-0.4) (P<0.001 by the Kruskal-Wallis test). FOXP3 mRNA levels were inversely correlated with serum creatinine levels measured at the time of biopsy in the acute-rejection group (Spearman's correlation coefficient = -0.38, P=0.02) but not in the group with chronic allograft nephropathy or the group with normal biopsy results. Analyses of receiver-operating-characteristic curves demonstrated that reversal of acute rejection can be predicted with 90 percent sensitivity and 73 percent specificity with use of the optimal identified cutoff for FOXP3 mRNA of 3.46 (P=0.001). FOXP3 mRNA levels identified subjects at risk for graft failure within six months after the incident episode of acute rejection (relative risk for the lowest third of FOXP3 mRNA levels, 6; P=0.02). None of the other mRNA levels were predictive of reversal of acute rejection or graft failure.


CONCLUSIONS
Measurement of FOXP3 mRNA in urine may offer a noninvasive means of improving the prediction of outcome of acute rejection of renal transplants.
Muthukumar
Thangamani
T
Division of Nephrology, Department of Medicine, Weill Medical College of Cornell University, New York, USA.
Dadhania
Darshana
D
Ding
Ruchuang
R
Snopkowski
Catherine
C
Naqvi
Rubina
R
Lee
Jun B
JB
Hartono
Choli
C
Li
Baogui
B
Sharma
Vijay K
VK
Seshan
Surya V
SV
Kapur
Sandip
S
Hancock
Wayne W
WW
Schwartz
Joseph E
JE
Suthanthiran
Manikkam
M
D000208:Acute Disease; D015415:Biomarkers / Q000097:blood / Q000652:urine; D017252:CD3 Complex / Q000652:urine; D003404:Creatinine / Q000097:blood; D051858:Forkhead Transcription Factors / Q000235:genetics* / Q000652:urine; D015870:Gene Expression; D006084:Graft Rejection / Q000097:blood / Q000175:diagnosis / Q000652:urine*; D006085:Graft Survival / Q000502:physiology; D006801:Humans; D016030:Kidney Transplantation; D008562:Membrane Glycoproteins / Q000652:urine; D054353:Perforin; D052899:Pore Forming Cytotoxic Proteins; D012333:RNA, Messenger / Q000652:urine*; D012337:RNA, Ribosomal, 18S / Q000652:urine; D012372:ROC Curve; D015375:Receptors, Interleukin-2 / Q000032:analysis; D012306:Risk; D013601:T-Lymphocytes / Q000276:immunology; D014184:Transplantation, Homologous
D015415:Biomarkers; D017252:CD3 Complex; C418974:FOXP3 protein, human; D051858:Forkhead Transcription Factors; D008562:Membrane Glycoproteins; D052899:Pore Forming Cytotoxic Proteins; D012333:RNA, Messenger; D012337:RNA, Ribosomal, 18S; D015375:Receptors, Interleukin-2; D054353:Perforin; D003404:Creatinine