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Prognostic cell-based assay predicts acute cellular rejection in pre- and post-transplant testing of children with liver and intestine transplantation (LTx, ITx).
27, Nov 2013

Prognostic cell-based assay predicts acute cellular rejection in pre- and post-transplant testing...

Background and Purpose: Children with LTx demonstrate lower rejection-risk and immunosuppression requirements compared with ITx recipients. Accurate rejection-risk assessment could enable early minimization and reduced drug toxicity in either recipient type. Allospecific CD154+T-cytotoxic memory cells (CD154+TcM) predict rejection in small cohorts of pediatric LTx and ITx and require validation

Methods: Between 2006-2012, screening-validation testing of CD154+TcM was undertaken at our center under protocol NCT#01163578 in children <21 years who receive LTx or ITx. In this cell-culture-based test, donor-specific immune response is expressed as the ratio of donor- and third-party-induced CD154+TcM. The ratio is termed immunoreactivity index or IR. In general, an IR > 1 implies increased donor-specific immune response and increased risk of rejection.

Results: 218 children provided 283 samples for testing. Donor- or third-party-induced CD154+TcM did not exceed background in 37 samples from 23 children and were discarded (Poisson statistic). A screening cohort of 147 samples from 120 subjects was tested with research-grade reagents and flow cytometers. In this cohort, an IR ≥ 1.1 in 98 post-transplant samples predicted rejection within 60 days after a test with performance (performance metrics are sensitivity, specificity, positive predictive value, and negative predictive values) of 92%, 84%, 65%, and 97%, respectively (AUC 0.878). In 49 pre-transplant samples, an IR ≥1.23 predicted rejection within the first 60 days after LTx or ITx with performance of 80%, 71%, 74%, and 77%, respectively (AUC 0.820). A validation cohort of 99 samples from 71 children was tested with a standardized test format using cGMP-manufactured reagents and FDA-approved instruments. Performance of the abovementioned pre- and post-transplant rejection-risk thresholds for predicting rejection was replicated in the validation cohort. Specifically, test performance in 67 post-transplant samples was 84%, 81%, 64%, and 93%, respectively (AUC 0.792). Test performance in 32 pre-transplant samples was 54%, 89%, 78%, and 74%, respectively (AUC 0.848).

Conclusions: In screening-replication testing of pre-transplant and post-transplant blood samples, allospecific CD154+T-cytotoxic memory cells predict acute cellular rejection after liver or intestine transplantation with high sensitivity and specificity. This performance can permit safe minimization of immunosuppression if confirmed in multicenter testing.


Plexision develops cellular biomarkers for personalized diagnosis and drug development in solid organ transplantation and immunological disorders. We also pioneer in R&D projects centered on integrating biomarker targets in all phases of drug development, from preclinical to post-marketing. Plexision’s technology can be adapted to

  • - Assess disease risk for several immunological disorders.
  • - Predict the success of a drug for a specific patient.
  • - Develop dosing recommendations for new immunological drugs.

Our state-of-the-art laboratory is CLIA-certified and located in Pittsburgh, PA.