Kidney and kidney-pancreas transplantationAn In Vitro System for the Determination of Individualized Immunosuppression
Section snippets
Patients and Methods
As a surrogate for the transplanted organ, donor lymphocytes were transformed by Epstein-Barr virus (EBV) subsequent to isolation. Recipient lymphocytes were isolated from a single blood specimen (10 to 20 mL) obtained after written informed consent was given by the patient. Blood collection was performed after preoperative immunosuppression was given (including 250 mg corticosteroids, 2 g mycophenolate mofetil, 5 mg/kg tacrolimus, and/or 0.2 mL/kg antithymocyte globuline). The study protocol
Results
Blood samples were obtained from 12 patients subsequent to organ transplantation (nine men, three women), mean age 44.3 years (range 27 to 72), having undergone transplantations of a kidney (n = 8), combined pancreas/kidney (n = 2), or a liver (n = 2). Lymphocyte isolation was successfully performed for all 12 blood samples. Seven EBV-transformed B cell lines (B-LCL) were established out of 11 lymphocyte preparations derived from organ donors (58.3%).
Using these cells, eight individual MLC
Discussion
As B cells of donor origin express both class I and class II HLA molecules and therefore resemble transplanted cells, they represent an appropriate target cell for this model of specific allogeneic stimulation. However, the analysis was hampered by the relatively poor yield of EBV transformation that must be improved for future analysis.
Specific alloreactivity could be demonstrated in all MLC pairs analyzed by ELISpot analysis. This method is well suited to measure alloreactivity as has been
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This work was supported by a FORUN grant of the Medical Faculty of the University of Rostock.
M.L. and C.R. contributed equally to this work.