Intragraft Expression of Transforming Growth Factor-Beta 1 by a Novel Quantitative Reverse Transcription Polymerase Chain Reaction Elisa in Long Lasting Kidney Recipients1

Diller, Ana; Albano, S; Giraudo, C; Alvarellos, Teresita; J, Sena; Massari, P.; G, de Boccardo

doi:10.1097/00007890-200008270-00014

Cited by 14 publications

(10 citation statements)

References 16 publications

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“…A second level of injury occurring after 12 months is associated with arteriolar hyalinosis, ischemic glomerulosclerosis, and secondary fibrosis, which is predominantly due to CNI toxicity (1,2). At the molecular level, there is increasing identification of chemokine, growth factor, and cytokine impact on tubulointerstitial damage (21)(22)(23). Other pathways include cellular senescence, epithelial-to-mesenchymal tubular cell transition, cytokine excess, and structural damage with loss of tubules leading to the formation of atubular glomeruli (24).…”

Section: Discussionmentioning

confidence: 99%

Molecular Pathways Involved in Loss of Kidney Graft Function with Tubular Atrophy and Interstitial Fibrosis

Maluf

Mas

Archer

et al. 2008

Mol Med

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Loss of kidney graft function with tubular atrophy (TA) and interstitial fibrosis (IF) causes most kidney allograft losses. We aimed to identify the molecular pathways involved in IF/TA progression. Kidney biopsies from normal kidneys (n = 24), normal allografts (n = 6), and allografts with IF/TA (n = 17) were analyzed using high-density oligonucleotide microarray. Probe set level tests of hypotheses tests were conducted to identify genes with a significant trend in gene expression across the three groups using Jonckheere-Terpstra test for trend. Interaction networks and functional analysis were used. An unsupervised hierarchical clustering analysis showed that all the IF/TA samples were associated with high correlation. Gene ontology classified the differentially expressed genes as related to immune response, inflammation, and matrix deposition. Chemokines (CX), CX receptor (for example, CCL5 and CXCR4), interleukin, and interleukin receptor (for example, IL-8 and IL10RA) genes were overexpressed in IF/TA samples compared with normal allografts and normal kidneys. Genes involved in apoptosis (for example, CASP4 and CASP5) were importantly overexpressed in IF/TA. Genes related to angiogenesis (for example, ANGPTL3, ANGPT2, and VEGF) were downregulated in IF/TA. Genes related to matrix production-deposition were upregulated in IF/TA. A distinctive gene expression pattern was observed in IF/TA samples compared with normal allografts and normal kidneys. We were able to establish a trend in gene expression for genes involved in different pathways among the studied groups. The top-scored networks were related to immune response, inflammation, and cellto-cell interaction, showing the importance of chronic inflammation in progressive graft deterioration.

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Section: Discussionmentioning

confidence: 99%

Molecular Pathways Involved in Loss of Kidney Graft Function with Tubular Atrophy and Interstitial Fibrosis

Maluf

Mas

Archer

et al. 2008

Mol Med

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“…Blood group genotypes are usually determined by electrophoresis of the amplicons in agarose gels. The main -TGT TCA ACA CCT ACT ATG CT Exon 5 125 Biotin-AAT ATG GAA AGC ACC TCA TG Exon 9 126 Biotin-AAG ACC TGT ACG CCA ACA CA b-Actin C/c 171 Biotin-(T) 12 TTC AGG GTG CCC TTT GTC ACT T Intron 2 (C) 170…”

Section: Discussionmentioning

confidence: 99%

“…Biotin-TCT ACG TGT TCG CAG CCT AT RHD/RHCE exon 4 K/k 180 Biotin-(T) 12 CGC ATC TCT GGT AAA T Exon 6 126…”

Section: Discussionmentioning

confidence: 99%

“…A less advanced, but more readily available, technique is the PCR-ELISA, in which digoxigenin-labeled amplicons are hybridized to a specific biotin-labeled oligonucleotide immobilized onto a microplate well via streptavidin. [12][13][14] The presence of digoxigenin-labeled amplicons can be detected with an anti-digoxigenin-conjugated reagent in a standard ELISA, similar to the ones already in use in blood banks to detect viral antibodies.…”

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confidence: 99%

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Extended blood grouping of blood donors with automatable PCR‐ELISA genotyping

2003

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“…Further, cross sectional studies of renal tissue in patients with established CAN have suggested that the development of glomerulosclerosis and interstitial fibrosis is mediated by the enhanced expression of the profibrotic growth factor, transforming growth factor-β (TGF-β) (13)(14)(15)(16)(17). Primary regulation of TGF-β activity is controlled by cleavage of the latent TGF-β precursor.…”

Section: Introductionmentioning

confidence: 99%

Evaluation of Gene Panel mRNAs in Urine Samples of Kidney Transplant Recipients as a Non-invasive Tool of Graft Function

Mas

Archer

et al. 2007

Mol Med

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Non-invasive monitoring may be useful after kidney transplantation (KT), particularly for predicting acute rejection (AR). It is less clear whether chronic allograft nephropathy (CAN) is also associated with changes in urine cells. To identify non-invasive markers of allograft function in kidney transplant patients (KTP), mRNA levels of AGT, TGF-β1, EGFR, IFN-γ, TSP-1, and IL-10 in urine (Ur) samples were studied using QRT-PCR. Ninety-five KTP and 111 Ur samples were evaluated. Patients (Pts) were divided as, within six months (N = 31), and with more than six months post-KT (N = 64). KTP with more than six months post-KT were classified as KTP with stable kidney function (SKF) (N = 32), KTP with SKF (creatinine < 2 mg/dL) and proteinuria > 500 mg/24 h (N = 18), and KTP with biopsy proven CAN (N = 14). F-test was used to test for equality of variances between groups. IL-10 mRNA was decreased in Ur samples from KTP with less than six months post-KT (P = 0.005). For KTR groups with more than six months post-KT, AGT and EGFR mRNA were statistically different among KTP with SKF, KTP with SKF and proteinuria, and CAN Pts (P = 0.003, and P = 0.01), with KTP with SKF having higher mean expression. TSP-1 mRNA levels also were significantly different among these three groups (P = 0.04), with higher expression observed in CAN Pts. Using the random forest algorithm, AGT, EGFR, and TGF-β1 were identified as predictors of CAN, SKF, SKF with proteinuria. A characteristic pattern of mRNA levels in the different KTP groups was observed indicating that the mRNA levels in Ur cells might reflect allograft function.

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Intragraft Expression of Transforming Growth Factor-Beta 1 by a Novel Quantitative Reverse Transcription Polymerase Chain Reaction Elisa in Long Lasting Kidney Recipients1

Abstract: This study showed that those patients with elevated CAN scores and higher proteinuria levels had higher TGF-beta1 intragraft expression.

Cited by 14 publications

References 16 publications

Molecular Pathways Involved in Loss of Kidney Graft Function with Tubular Atrophy and Interstitial Fibrosis

Molecular Pathways Involved in Loss of Kidney Graft Function with Tubular Atrophy and Interstitial Fibrosis

Extended blood grouping of blood donors with automatable PCR‐ELISA genotyping

Evaluation of Gene Panel mRNAs in Urine Samples of Kidney Transplant Recipients as a Non-invasive Tool of Graft Function

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