Intragraft expression of transforming growth factor-beta 1 by a novel quantitative reverse transcription polymerase chain reaction elisa in long lasting kidney recipients

Valeria Mas, Ana Diller, Susana Albano, Constancio Giraudo, Teresita Alvarellos, Javier Sena, Pablo Massari, Graciela De Boccardo

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

Background. Chronic allograft nephropathy (CAN) remains a major problem in clinical transplantation. It has been associated with increased transforming growth factor (TGF-β1). Our goal was to correlate CAN and levels of TGF-β1 by using a novel competitive quantitative for reverse transcription-polymerase chain reaction-ELISA (RT-PCR-ELISA) assay. Methods. We studied 12 transplantation patients (posttransplant time: 36.5±11.2 months, range (r): 13-52) with stable creatinine and blood pressure and varled proteinuria. A Kidney biopsy was performed in all patients. Six patients with acute tubular necrosis (ATN) immediately after transplantation were used as controls. Histopathological evaluation was based on Banff working classification criteria. We designed an heterologous RNA competitor (IC) for RT-PCR-ELISA, which co-amplified with the same primer as TGF-β1. Products were viewed on 96-well plates labeled with probes for IC at the desired sequence. Results. Results were expressed as the number of TGF-β1 copies/μg of total RNA. Six patients showed more than 1000 mg/24 hr proteinuria (2446±1421 mg/24 hr, r: 1200-5000) higher CAN Banff scores, and the other six presented <1,000 mg/24 hr (348±267 mg/24 hr, r: 114-800). This difference was significant (P=0.01). There were not significant differences in posttransplant time, creatinine, or blood pressure between groups. TGF-β1 levels by RT-PCR-ELISA were statistically significant (6038±5317, r: 1239-12100 versus 177±119.7, r: 51-400, P=0.04). The control group showed levels of 228±111, r: 140-444, P=0.04) with significant difference only for the higher proteinuria group (P=0.03). Conclusions. This study showed that those patients with elevated CAN scores and higher proteinuria levels had higher TGF-β1 intragraft expression.

Original languageEnglish (US)
Pages (from-to)612-616
Number of pages5
JournalTransplantation
Volume70
Issue number4
StatePublished - Aug 27 2000
Externally publishedYes

Fingerprint

Transforming Growth Factor beta
Reverse Transcription
Proteinuria
Allografts
Kidney
Polymerase Chain Reaction
Transplantation
Enzyme-Linked Immunosorbent Assay
Creatinine
RNA
Blood Pressure
Transforming Growth Factors
Necrosis
Biopsy
Control Groups

ASJC Scopus subject areas

  • Transplantation
  • Immunology

Cite this

Intragraft expression of transforming growth factor-beta 1 by a novel quantitative reverse transcription polymerase chain reaction elisa in long lasting kidney recipients. / Mas, Valeria; Diller, Ana; Albano, Susana; Giraudo, Constancio; Alvarellos, Teresita; Sena, Javier; Massari, Pablo; De Boccardo, Graciela.

In: Transplantation, Vol. 70, No. 4, 27.08.2000, p. 612-616.

Research output: Contribution to journalArticle

Mas, V, Diller, A, Albano, S, Giraudo, C, Alvarellos, T, Sena, J, Massari, P & De Boccardo, G 2000, 'Intragraft expression of transforming growth factor-beta 1 by a novel quantitative reverse transcription polymerase chain reaction elisa in long lasting kidney recipients', Transplantation, vol. 70, no. 4, pp. 612-616.
Mas, Valeria ; Diller, Ana ; Albano, Susana ; Giraudo, Constancio ; Alvarellos, Teresita ; Sena, Javier ; Massari, Pablo ; De Boccardo, Graciela. / Intragraft expression of transforming growth factor-beta 1 by a novel quantitative reverse transcription polymerase chain reaction elisa in long lasting kidney recipients. In: Transplantation. 2000 ; Vol. 70, No. 4. pp. 612-616.
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abstract = "Background. Chronic allograft nephropathy (CAN) remains a major problem in clinical transplantation. It has been associated with increased transforming growth factor (TGF-β1). Our goal was to correlate CAN and levels of TGF-β1 by using a novel competitive quantitative for reverse transcription-polymerase chain reaction-ELISA (RT-PCR-ELISA) assay. Methods. We studied 12 transplantation patients (posttransplant time: 36.5±11.2 months, range (r): 13-52) with stable creatinine and blood pressure and varled proteinuria. A Kidney biopsy was performed in all patients. Six patients with acute tubular necrosis (ATN) immediately after transplantation were used as controls. Histopathological evaluation was based on Banff working classification criteria. We designed an heterologous RNA competitor (IC) for RT-PCR-ELISA, which co-amplified with the same primer as TGF-β1. Products were viewed on 96-well plates labeled with probes for IC at the desired sequence. Results. Results were expressed as the number of TGF-β1 copies/μg of total RNA. Six patients showed more than 1000 mg/24 hr proteinuria (2446±1421 mg/24 hr, r: 1200-5000) higher CAN Banff scores, and the other six presented <1,000 mg/24 hr (348±267 mg/24 hr, r: 114-800). This difference was significant (P=0.01). There were not significant differences in posttransplant time, creatinine, or blood pressure between groups. TGF-β1 levels by RT-PCR-ELISA were statistically significant (6038±5317, r: 1239-12100 versus 177±119.7, r: 51-400, P=0.04). The control group showed levels of 228±111, r: 140-444, P=0.04) with significant difference only for the higher proteinuria group (P=0.03). Conclusions. This study showed that those patients with elevated CAN scores and higher proteinuria levels had higher TGF-β1 intragraft expression.",
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T1 - Intragraft expression of transforming growth factor-beta 1 by a novel quantitative reverse transcription polymerase chain reaction elisa in long lasting kidney recipients

AU - Mas, Valeria

AU - Diller, Ana

AU - Albano, Susana

AU - Giraudo, Constancio

AU - Alvarellos, Teresita

AU - Sena, Javier

AU - Massari, Pablo

AU - De Boccardo, Graciela

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N2 - Background. Chronic allograft nephropathy (CAN) remains a major problem in clinical transplantation. It has been associated with increased transforming growth factor (TGF-β1). Our goal was to correlate CAN and levels of TGF-β1 by using a novel competitive quantitative for reverse transcription-polymerase chain reaction-ELISA (RT-PCR-ELISA) assay. Methods. We studied 12 transplantation patients (posttransplant time: 36.5±11.2 months, range (r): 13-52) with stable creatinine and blood pressure and varled proteinuria. A Kidney biopsy was performed in all patients. Six patients with acute tubular necrosis (ATN) immediately after transplantation were used as controls. Histopathological evaluation was based on Banff working classification criteria. We designed an heterologous RNA competitor (IC) for RT-PCR-ELISA, which co-amplified with the same primer as TGF-β1. Products were viewed on 96-well plates labeled with probes for IC at the desired sequence. Results. Results were expressed as the number of TGF-β1 copies/μg of total RNA. Six patients showed more than 1000 mg/24 hr proteinuria (2446±1421 mg/24 hr, r: 1200-5000) higher CAN Banff scores, and the other six presented <1,000 mg/24 hr (348±267 mg/24 hr, r: 114-800). This difference was significant (P=0.01). There were not significant differences in posttransplant time, creatinine, or blood pressure between groups. TGF-β1 levels by RT-PCR-ELISA were statistically significant (6038±5317, r: 1239-12100 versus 177±119.7, r: 51-400, P=0.04). The control group showed levels of 228±111, r: 140-444, P=0.04) with significant difference only for the higher proteinuria group (P=0.03). Conclusions. This study showed that those patients with elevated CAN scores and higher proteinuria levels had higher TGF-β1 intragraft expression.

AB - Background. Chronic allograft nephropathy (CAN) remains a major problem in clinical transplantation. It has been associated with increased transforming growth factor (TGF-β1). Our goal was to correlate CAN and levels of TGF-β1 by using a novel competitive quantitative for reverse transcription-polymerase chain reaction-ELISA (RT-PCR-ELISA) assay. Methods. We studied 12 transplantation patients (posttransplant time: 36.5±11.2 months, range (r): 13-52) with stable creatinine and blood pressure and varled proteinuria. A Kidney biopsy was performed in all patients. Six patients with acute tubular necrosis (ATN) immediately after transplantation were used as controls. Histopathological evaluation was based on Banff working classification criteria. We designed an heterologous RNA competitor (IC) for RT-PCR-ELISA, which co-amplified with the same primer as TGF-β1. Products were viewed on 96-well plates labeled with probes for IC at the desired sequence. Results. Results were expressed as the number of TGF-β1 copies/μg of total RNA. Six patients showed more than 1000 mg/24 hr proteinuria (2446±1421 mg/24 hr, r: 1200-5000) higher CAN Banff scores, and the other six presented <1,000 mg/24 hr (348±267 mg/24 hr, r: 114-800). This difference was significant (P=0.01). There were not significant differences in posttransplant time, creatinine, or blood pressure between groups. TGF-β1 levels by RT-PCR-ELISA were statistically significant (6038±5317, r: 1239-12100 versus 177±119.7, r: 51-400, P=0.04). The control group showed levels of 228±111, r: 140-444, P=0.04) with significant difference only for the higher proteinuria group (P=0.03). Conclusions. This study showed that those patients with elevated CAN scores and higher proteinuria levels had higher TGF-β1 intragraft expression.

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