Noninvasive diagnosis of renal-allograft rejection by measurement of messenger RNA for perforin and granzyme B in urine

Baogui Li, Choli Hartono, Ruchuang Ding, Vijay K. Sharma, Ravi Ramaswamy, Biao Qian, David Serur, Janet Mouradian, Joseph E. Schwartz, Manikkam Suthanthiran

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Abstract

Background: Acute rejection is a serious and frequent complication of renal transplantation, and its diagnosis is contingent on the invasive procedure of allograft biopsy. A noninvasive diagnostic test for rejection could improve the outcome of transplantation. Methods: We obtained 24 urine specimens from 22 renal-allograft recipients with a biopsy-confirmed episode of acute rejection and 127 samples from 63 recipients without evidence of acute rejection. RNA was isolated from the urinary cells. Messenger RNA (mRNA) encoding the cytotoxic proteins perforin and granzyme B and a constitutively expressed cyclophilin B gene were measured with the use of a competitive, quantitative polymerase-chain-reaction assay, and the level of expression was correlated with allograft status. Results: The log-transformed mean (±SE) levels of perforin mRNA and granzyme B mRNA, which encode cytotoxic proteins, but not the levels of constitutively expressed cyclophilin B mRNA, were higher in the urinary cells from the 22 patients with a biopsy-confirmed episode of acute rejection than in the 63 recipients without an episode of acute rejection (perforin, 1.4±0.3 vs. -0.6±0.2 fg per microgram of total RNA; P<0.001; and granzyme B, 1.2±0.3 vs. -0.9±0.2 fg per microgram of total RNA; P<0.001). Analysis involving the receiver-operating-characteristic curve demonstrated that acute rejection can be predicted with a sensitivity of 83 percent and a specificity of 83 percent with the use of a cutoff value of 0.9 fg of perforin mRNA per microgram of total RNA, and with a sensitivity of 79 percent and a specificity of 77 percent with the use of a cutoff value of 0.4 fg of granzyme B mRNA per microgram of total RNA. Sequential urine samples were obtained from 37 patients during the first nine days after transplantation, and measurements of the levels of mRNA that encoded cytotoxic proteins identified those in whom acute rejection developed. Conclusions: Measurement of mRNA encoding cytotoxic proteins in urinary cells offers a noninvasive means of diagnosing acute rejection of renal allografts.

Original languageEnglish
Pages (from-to)947-954
Number of pages8
JournalNew England Journal of Medicine
Volume344
Issue number13
DOIs
Publication statusPublished - 29 Mar 2001
Externally publishedYes

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Allografts
Urine
Kidney
Messenger RNA
Granzymes
Perforin
RNA
Biopsy
Transplantation
Proteins
perforin-granzyme B
Routine Diagnostic Tests
ROC Curve
Kidney Transplantation
Polymerase Chain Reaction
Genes

ASJC Scopus subject areas

  • Medicine(all)

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Noninvasive diagnosis of renal-allograft rejection by measurement of messenger RNA for perforin and granzyme B in urine. / Li, Baogui; Hartono, Choli; Ding, Ruchuang; Sharma, Vijay K.; Ramaswamy, Ravi; Qian, Biao; Serur, David; Mouradian, Janet; Schwartz, Joseph E.; Suthanthiran, Manikkam.

In: New England Journal of Medicine, Vol. 344, No. 13, 29.03.2001, p. 947-954.

Research output: Contribution to journalArticle

Li, B, Hartono, C, Ding, R, Sharma, VK, Ramaswamy, R, Qian, B, Serur, D, Mouradian, J, Schwartz, JE & Suthanthiran, M 2001, 'Noninvasive diagnosis of renal-allograft rejection by measurement of messenger RNA for perforin and granzyme B in urine', New England Journal of Medicine, vol. 344, no. 13, pp. 947-954. https://doi.org/10.1056/NEJM200103293441301
Li, Baogui ; Hartono, Choli ; Ding, Ruchuang ; Sharma, Vijay K. ; Ramaswamy, Ravi ; Qian, Biao ; Serur, David ; Mouradian, Janet ; Schwartz, Joseph E. ; Suthanthiran, Manikkam. / Noninvasive diagnosis of renal-allograft rejection by measurement of messenger RNA for perforin and granzyme B in urine. In: New England Journal of Medicine. 2001 ; Vol. 344, No. 13. pp. 947-954.
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abstract = "Background: Acute rejection is a serious and frequent complication of renal transplantation, and its diagnosis is contingent on the invasive procedure of allograft biopsy. A noninvasive diagnostic test for rejection could improve the outcome of transplantation. Methods: We obtained 24 urine specimens from 22 renal-allograft recipients with a biopsy-confirmed episode of acute rejection and 127 samples from 63 recipients without evidence of acute rejection. RNA was isolated from the urinary cells. Messenger RNA (mRNA) encoding the cytotoxic proteins perforin and granzyme B and a constitutively expressed cyclophilin B gene were measured with the use of a competitive, quantitative polymerase-chain-reaction assay, and the level of expression was correlated with allograft status. Results: The log-transformed mean (±SE) levels of perforin mRNA and granzyme B mRNA, which encode cytotoxic proteins, but not the levels of constitutively expressed cyclophilin B mRNA, were higher in the urinary cells from the 22 patients with a biopsy-confirmed episode of acute rejection than in the 63 recipients without an episode of acute rejection (perforin, 1.4±0.3 vs. -0.6±0.2 fg per microgram of total RNA; P<0.001; and granzyme B, 1.2±0.3 vs. -0.9±0.2 fg per microgram of total RNA; P<0.001). Analysis involving the receiver-operating-characteristic curve demonstrated that acute rejection can be predicted with a sensitivity of 83 percent and a specificity of 83 percent with the use of a cutoff value of 0.9 fg of perforin mRNA per microgram of total RNA, and with a sensitivity of 79 percent and a specificity of 77 percent with the use of a cutoff value of 0.4 fg of granzyme B mRNA per microgram of total RNA. Sequential urine samples were obtained from 37 patients during the first nine days after transplantation, and measurements of the levels of mRNA that encoded cytotoxic proteins identified those in whom acute rejection developed. Conclusions: Measurement of mRNA encoding cytotoxic proteins in urinary cells offers a noninvasive means of diagnosing acute rejection of renal allografts.",
author = "Baogui Li and Choli Hartono and Ruchuang Ding and Sharma, {Vijay K.} and Ravi Ramaswamy and Biao Qian and David Serur and Janet Mouradian and Schwartz, {Joseph E.} and Manikkam Suthanthiran",
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T1 - Noninvasive diagnosis of renal-allograft rejection by measurement of messenger RNA for perforin and granzyme B in urine

AU - Li, Baogui

AU - Hartono, Choli

AU - Ding, Ruchuang

AU - Sharma, Vijay K.

AU - Ramaswamy, Ravi

AU - Qian, Biao

AU - Serur, David

AU - Mouradian, Janet

AU - Schwartz, Joseph E.

AU - Suthanthiran, Manikkam

PY - 2001/3/29

Y1 - 2001/3/29

N2 - Background: Acute rejection is a serious and frequent complication of renal transplantation, and its diagnosis is contingent on the invasive procedure of allograft biopsy. A noninvasive diagnostic test for rejection could improve the outcome of transplantation. Methods: We obtained 24 urine specimens from 22 renal-allograft recipients with a biopsy-confirmed episode of acute rejection and 127 samples from 63 recipients without evidence of acute rejection. RNA was isolated from the urinary cells. Messenger RNA (mRNA) encoding the cytotoxic proteins perforin and granzyme B and a constitutively expressed cyclophilin B gene were measured with the use of a competitive, quantitative polymerase-chain-reaction assay, and the level of expression was correlated with allograft status. Results: The log-transformed mean (±SE) levels of perforin mRNA and granzyme B mRNA, which encode cytotoxic proteins, but not the levels of constitutively expressed cyclophilin B mRNA, were higher in the urinary cells from the 22 patients with a biopsy-confirmed episode of acute rejection than in the 63 recipients without an episode of acute rejection (perforin, 1.4±0.3 vs. -0.6±0.2 fg per microgram of total RNA; P<0.001; and granzyme B, 1.2±0.3 vs. -0.9±0.2 fg per microgram of total RNA; P<0.001). Analysis involving the receiver-operating-characteristic curve demonstrated that acute rejection can be predicted with a sensitivity of 83 percent and a specificity of 83 percent with the use of a cutoff value of 0.9 fg of perforin mRNA per microgram of total RNA, and with a sensitivity of 79 percent and a specificity of 77 percent with the use of a cutoff value of 0.4 fg of granzyme B mRNA per microgram of total RNA. Sequential urine samples were obtained from 37 patients during the first nine days after transplantation, and measurements of the levels of mRNA that encoded cytotoxic proteins identified those in whom acute rejection developed. Conclusions: Measurement of mRNA encoding cytotoxic proteins in urinary cells offers a noninvasive means of diagnosing acute rejection of renal allografts.

AB - Background: Acute rejection is a serious and frequent complication of renal transplantation, and its diagnosis is contingent on the invasive procedure of allograft biopsy. A noninvasive diagnostic test for rejection could improve the outcome of transplantation. Methods: We obtained 24 urine specimens from 22 renal-allograft recipients with a biopsy-confirmed episode of acute rejection and 127 samples from 63 recipients without evidence of acute rejection. RNA was isolated from the urinary cells. Messenger RNA (mRNA) encoding the cytotoxic proteins perforin and granzyme B and a constitutively expressed cyclophilin B gene were measured with the use of a competitive, quantitative polymerase-chain-reaction assay, and the level of expression was correlated with allograft status. Results: The log-transformed mean (±SE) levels of perforin mRNA and granzyme B mRNA, which encode cytotoxic proteins, but not the levels of constitutively expressed cyclophilin B mRNA, were higher in the urinary cells from the 22 patients with a biopsy-confirmed episode of acute rejection than in the 63 recipients without an episode of acute rejection (perforin, 1.4±0.3 vs. -0.6±0.2 fg per microgram of total RNA; P<0.001; and granzyme B, 1.2±0.3 vs. -0.9±0.2 fg per microgram of total RNA; P<0.001). Analysis involving the receiver-operating-characteristic curve demonstrated that acute rejection can be predicted with a sensitivity of 83 percent and a specificity of 83 percent with the use of a cutoff value of 0.9 fg of perforin mRNA per microgram of total RNA, and with a sensitivity of 79 percent and a specificity of 77 percent with the use of a cutoff value of 0.4 fg of granzyme B mRNA per microgram of total RNA. Sequential urine samples were obtained from 37 patients during the first nine days after transplantation, and measurements of the levels of mRNA that encoded cytotoxic proteins identified those in whom acute rejection developed. Conclusions: Measurement of mRNA encoding cytotoxic proteins in urinary cells offers a noninvasive means of diagnosing acute rejection of renal allografts.

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