title: Predicting Cellular Rejection With a Cell-Based Assay: Preclinical Evaluation in Children.
abstract: BACKGROUND
  Allospecific CD154+T-cytotoxic memory cells (CD154+TcM) predict acute cellular rejection after liver transplantation (LTx) or intestine transplantation (ITx) in small cohorts of children and can enhance immunosuppression management, but await validation and clinical implementation.
  
  
  METHODS
  To establish safety and probable benefit, CD154+TcM were measured in cryopreserved samples from 214 children younger than 21 years (National Clinical Trial 1163578). Training set samples (n = 158) were tested with research-grade reagents and 122 independent validation set samples were tested with current good manufacturing practices-manufactured reagents after assay standardization and reproducibility testing. Recipient CD154+TcM induced by stimulation with donor cells were expressed as a fraction of those induced by HLA nonidentical cells in parallel cultures. The resulting immunoreactivity index (IR) if greater than 1 implies increased rejection-risk.
  
  
  RESULTS
  Training and validation set subjects were demographically similar. Mean coefficient of test variation was less than 10% under several conditions. Logistic regression incorporating several confounding variables identified separate pretransplant and posttransplant IR thresholds for prediction of rejection in the respective training set samples. An IR of 1.1 or greater in posttransplant training samples and IR of 1.23 or greater in pretransplant training samples predicted LTx or ITx rejection in corresponding validation set samples in the 60-day postsampling period with sensitivity, specificity, positive, and negative predictive values of 84%, 80%, 64%, and 92%, respectively (area under the receiver operator characteristic curve, 0.792), and 57%, 89%, 78%, and 74%, respectively (area under the receiver operator characteristic curve, 0.848). No adverse events were encountered due to phlebotomy.
  
  
  CONCLUSIONS
  Allospecific CD154+T-cytotoxic memory cells predict acute cellular rejection after LTx or ITx in children. Adjunctive use can enhance clinical outcomes.
authors.affiliation: 1 Thomas E. Starzl Transplantation Institute, Hillman Center for Pediatric Transplantation, Department of Transplant Surgery, Children's Hospital of Pittsburgh of University of Pittsburgh Medical Center, Pittsburgh, PA. 2 Plexision Inc, Pittsburgh, PA. 3 Tissue Typing Laboratory, Department of Pathology, Children's Hospital of Pittsburgh of University of Pittsburgh Medical Center, Pittsburgh, PA.
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authors.forename: Brandon W
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authors.lastname: Higgs
authors.forename: Mylarappa
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authors.forename: Amanda
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authors.forename: Jaimie
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authors.lastname: White
authors.forename: Samantha
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authors.lastname: Fazzolare
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authors.lastname: Remaley
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authors.lastname: Dirling
authors.forename: Patricia
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authors.lastname: Harris
authors.forename: Tara
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authors.lastname: Hartle
authors.forename: Pamela
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authors.lastname: Kachmar
authors.forename: Megan
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authors.lastname: Nicely
authors.forename: Lindsay
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authors.lastname: OʼToole
authors.forename: Brittany
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authors.lastname: Boehm
authors.forename: Nicole
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authors.lastname: Jativa
authors.forename: Paula
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authors.lastname: Stanley
authors.forename: Ronald
authors.initials: R
authors.lastname: Jaffe
authors.forename: Sarangarajan
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authors.lastname: Ranganathan
authors.forename: Adriana
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authors.lastname: Zeevi
authors.forename: Rakesh
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authors.lastname: Sindhi
mesh_terms: D000208:Acute Disease; D000293:Adolescent; D019540:Area Under Curve; D015415:Biomarkers / Q000032:analysis; D023201:CD40 Ligand / Q000032:analysis*; D002478:Cells, Cultured; D002648:Child; D002675:Child, Preschool; D015925:Cryopreservation; D005260:Female; D005434:Flow Cytometry; D006084:Graft Rejection / Q000276:immunology* / Q000517:prevention & control; D006801:Humans; D007111:Immunity, Cellular / Q000187:drug effects; D007156:Immunologic Memory; D007159:Immunologic Tests / Q000379:methods*; D007166:Immunosuppressive Agents / Q000627:therapeutic use; D007223:Infant; D007422:Intestines / Q000276:immunology / Q000637:transplantation*; D016031:Liver Transplantation / Q000009:adverse effects*; D016015:Logistic Models; D008297:Male; D015999:Multivariate Analysis; D011237:Predictive Value of Tests; D012372:ROC Curve; D015203:Reproducibility of Results; D018570:Risk Assessment; D012307:Risk Factors; D013602:T-Lymphocytes, Cytotoxic / Q000187:drug effects / Q000276:immunology*; D013997:Time Factors; D016896:Treatment Outcome; D055815:Young Adult
chemical_list: D015415:Biomarkers; D007166:Immunosuppressive Agents; D023201:CD40 Ligand