APPLICATION OF IN-SITU HYBRIDIZATION TECHNIQUE FOR QUANTITATIVE ASSESSMENT OF ONGOING SYMPTOMATIC EPSTEIN-BARR-VIRUS INFECTION AFTER LIVING-RELATED LIVER-TRANSPLANTATION

For quantitative assessment of ongoing symptomatic Epstein-Barr virus (EBV) infection in pediatric recipients of liver transplantation, we d etermined the number of peripheral blood mononuclear cells (PBMC) infe cted by EBV by in situ hybridization (ISH) and related the results wit h clinical courses of those patients. Twenty-four patients had symptom atic EBV infection between February 1995 and March 1996. Blood samples were obtained from these 24 patients at the time of acute phase, from 13 of them during convalescence, and 37 pediatric patients before tra nsplantation. ISH was performed on the PBMC and polymerase chain react ion (PCR) on DNA from whole blood. Oligonucleotide probes for ISH were chosen from coding sequences of EBV-encoded small nuclear RNA 1 (EBER 1). Results of ISH were reported in a number of cells expressing EBER1 /5 x 104 PBMC (# EBER1). Fever, diarrhea, upper respiratory symptoms, pleural effusion, ascites, lymphadenopathy, and lymphoproliferative di sease (LPD) accompanied with EBV infection proven by serology, viral-s pecific stain or PCR were regarded as EBV related diseases (EBVD), All samples with positive # EBER1 were accompanied by positive EBV PCR. # EBER1 was 68.2 +/- 144.9 (mean +/- SD) ranging from 0 to 621 in the a cute phase, 0.20 +/- 0.41 ranging from 0 to 2 in the convalescence pha se, 0.27 +/- 0.77 in 23 preoperative patients with positive serology, and 0 in all 14 preoperative patients with negative serology, The # EB ER1 in ongoing EBVD was significantly greater than that of patients in convalescence or before transplantation. Patients with # EBER1 greate r than 10 had a significantly lower chance of convalescence and a high er mortality than patients with # EBER1 less than 10. We conclude that # EBER1 could be a specific and quantitative marker of EBVD and might predict progression to LPD.