title: Use of Quantitative Real Time Polymerase Chain Reaction to Assess Gene Transcripts Associated With Antibody-Mediated Rejection of Kidney Transplants.
abstract: BACKGROUND
  Microarray studies have shown elevated transcript levels of endothelial and natural killer (NK) cell-associated genes during antibody-mediated rejection (AMR) of the renal allograft. This study aimed to assess the use of quantitative real-time polymerase chain reaction as an alternative to microarray analysis on a subset of these elevated genes.
  
  
  METHODS
  Thirty-nine renal transplant biopsies from patients with de novo donor-specific antibodies and eighteen 1-year surveillance biopsies with no histological evidence of rejection were analyzed for expression of 11 genes previously identified as elevated in AMR.
  
  
  RESULTS
  Expression levels of natural killer markers were correlated to microcirculation inflammation and graft outcomes to a greater extent than endothelial markers. Creating a predictive model reduced the number of gene transcripts to be assessed to 2, SH2D1b and MYBL1, resulting in 66.7% sensitivity and 89.7% specificity for graft loss.
  
  
  CONCLUSIONS
  This work demonstrates that elevated gene expression levels, proposed to be associated with AMR, can be detected by established quantitative real-time polymerase chain reaction technology, making transition to the clinical setting feasible. Transcript analysis provides additional diagnostic information to the classification schema for AMR diagnosis but it remains to be determined whether significant numbers of centres will validate transcript analysis in their laboratories and put such analysis into clinical use.
authors: 1 Centre for Complement and Inflammation Research, Division of Immunology and Inflammation, Department of Medicine, Imperial College, London, United Kingdom. 2 Department of Cellular Pathology, Hammersmith Hospital, London, United Kingdom. 3 Imperial College Kidney and Transplant Institute, Hammersmith Hospital, London, United Kingdom. 4 Histocompatibility and Immunogenetics Laboratory, Imperial College Healthcare NHS Trust, London, United Kingdom. 5 Medical Research Council Clinical Sciences Centre, Faculty of Medicine, Imperial College, London, United Kingdom.
authors: Katherine M
authors: KM
authors: Dominy
authors: Candice
authors: C
authors: Roufosse
authors: Hanneke
authors: H
authors: de Kort
authors: Michelle
authors: M
authors: Willicombe
authors: Paul
authors: P
authors: Brookes
authors: Jacques V
authors: JV
authors: Behmoaras
authors: Enrico G
authors: EG
authors: Petretto
authors: Jack
authors: J
authors: Galliford
authors: Peter
authors: P
authors: Choi
authors: David
authors: D
authors: Taube
authors: H Terence
authors: HT
authors: Cook
authors: Adam G
authors: AG
authors: Mclean
mesh_terms: D001706:Biopsy; D016022:Case-Control Studies; D020869:Gene Expression Profiling / Q000379:methods*; D005819:Genetic Markers; D006084:Graft Rejection / Q000235:genetics* / Q000276:immunology / Q000401:mortality / Q000473:pathology; D006085:Graft Survival; D006801:Humans; D056724:Immunity, Humoral / Q000187:drug effects*; D053208:Kaplan-Meier Estimate; D016030:Kidney Transplantation / Q000009:adverse effects* / Q000401:mortality; D011237:Predictive Value of Tests; D011518:Proto-Oncogene Proteins / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction; D012307:Risk Factors; D015534:Trans-Activators / Q000235:genetics; D014157:Transcription Factors / Q000235:genetics; D014158:Transcription, Genetic; D016896:Treatment Outcome
chemical_list: D005819:Genetic Markers; C088831:MYBL1 protein, human; D011518:Proto-Oncogene Proteins; D012333:RNA, Messenger; C103572:SH2D1B protein, human; D015534:Trans-Activators; D014157:Transcription Factors