Cytokine profile in graft-versus-host disease after small bowel transplantation.

Male ACI (Rna) and Lewis (LEW. Rnl) rats were used as donors and recipients. respectively. Simultaneous orthotopic small bowel transplantation (portocaval drainage) with bone marrow (2.5 X lOS unfractionated cells) infusion was perfonned under immunosuppression of intramuscular FK 506 1.0 mglkgld for 2 weeks (day 0 to 13), followed by a weekly injection of the same dose thereafter. All recipients developed clinical signs of GVHD (skin rash. pile loss. and hyperkeratosis) between 98 and 107 days after transplantation. Recipients were sacrificed after clinical manifestation of GVHD (120 days after transplantation. group 1. n = 3) or prior to any overt sign of GVHD (65 days after transplantation. group 2. n = 3). Controls comprised FK 506 treated Lewis recipients of isografts. sacrificed either 120 days (group 3. n = 3) or 65 days (group 4. n = 3) after transplantation. At the time of sacrifice. the cervical lymph nodes (CLN). tongue (TO). graft (GSB), and recipient small bowel (RSB) were harvested for histopathologic examination and semiquantitative cytokine analysis by reverse transcription polymerase chain reaction (RT-PCR) using 32p terminally labeled primer for interleukin (IL)-2. IL-4. IL-I0. and interferon (IFN)_y.2 f3-actin level in each sample was assessed for internal quality control and results were expressed as a ratio of cytokine/f3-actin mRNA.