Proteomic Screening Applied To Early The Diagnosis Of Chronic Graft-Versus-Host-Disease

Chronic graft versus host disease (GvHD), or late acute GvHD play a major role in morbidity, but also mortality of patients post allogeneic HSCT. Diagnosis chronic GvHD is based on clinical features and biopsies, a non invasive, unbiased laboratory test does not exist. We established a proteomic screening test for cGvHD by using capillary electrophoresis (CE) coupled on line to an ESI-Tof (electrospray-ionization time of flight) mass spectrometer (MS). Screening urine collected from 20 patients (10 with limited cGvHD, 10 with extensive cGvHD) we established a proteomic pattern allowing the diagnosis of cGvHD development. The resulting set of polypeptide markers (27 differentially excreted peptides) was tested on more than 227 patients prospectively and blinded for the correct classification of cGvHD samples. The majority of the patients included were transplanted for hematological malignancies (n=209), 6 for hematopoietic failure syndromes. The cGvHD proteomic pattern of 27 peptides was established by using 20 cases and 20 controls, consisting of patients at least 100 days post-HSCT, with no GvHD in the history, no infections and without relapse at the time of sampling. Aproteomic pattern differentiating chronic from acute GvHD was developed, since the initial pattern did cross react with aGvHD patients.Prospective and blinded evaluation of the patients revealed the correct classification of patients developing cGvHD with a sensitivity of 85% and specificity of 95%. Interestingly, urine from patients developing GvHD post DLI did react with the cGvHD-pattern and is predictive for GvHD developing post-DLI (about 2 weeks prior to DLI-GvHD development When the aGvHD-specific proteomic pattern was applied to the screening of patients with DLI in order to predict GvHD, only GvHD of the intestine reacted significantly with the aGvHD pattern. Further evaluations of the cGvHD patterns specific for particular organ manifestations are currently ongoing.