Influence of Back Table Portal and Arterial Flushing with HTK and Tacrolimus on the Incidence of Liver Graft Dysfunction P-702 A Shcherba 1, A Minou 2, D Efimov 1, S Korotkov 1, O Lebedz 3, A Dzyadzko 2, A Karytka 4, E Santotski 2 and Oleg O Rummo 1 1 Department of Transplantology, 2 Department of Anesthesiology and Intensive Care, 4 Cell Biotechnology Laboratory, RSP Center for Organ and Tissue Transplantation; 3 Department of Histology, Pathology Bureau, Minsk, Belarus OBJECTIVE To evaluate the possibility of back-table liver flushing with high exposure to tacrolimus to attenuate the early allograft dysfunction (EAD). Conflict of interests: none to declare CONCLUSION Preliminary data show that back-table liver flushing with high exposure to tacrolimus may contribute to lower EAD incidence. The possible mechanism of action may involve downregulation of IL-17 and inhibition of liver CD68 expression early post reperfusion. RESULTS No diffference was found between study groups (main vs control) regarding steatosis (16.9 vs 19.1; p=0.7), ballooning (35 vs 42; p=0.4) of liver grafts, donor age (44,5 vs 39,5;p=0.08), warm ischemia time (50 vs 50;p=0.5) and total ischemia time (482 vs 490;p=0.6). The rate of EAD was significantly lower in the main group (6/38 vs 15/34; Fisher test, p=0,01). There were one case of PNF in each group (1/38 vs 1/34; p=1). The median highest level of AST 24 h after reperfusion (1008 vs 1705;p=0,001) and ALT 24 h after reperfusion (452 vs 869;p=0,04) were significantly lower in the intervention group comparing to control group. The median highest levels of AST and ALT 48 h after reperfusion were not different between study groups (467 vs 641, p=0.15; 552 vs 667, p=0,30). There was no difference in hyperfibrinolysis rate (4/38 vs 6/34; p=0,5), the rate of septic complications (9/38 vs 6/34;p=0.5) and need for renal replacement therapy (5/38 vs 3/34;p=0.7) between the study groups. Hospital mortality (1/38 vs 1/34;p=1) was similar in both groups. In order to investigate the reason of difference of EAD incidence between groups we studied the levels of IL-6, 8, 17, 23, TNF-a and MIP-1a in samples 0, 1 and 2 and P-selectin in samples 0 and 1 in 26 patients (13 in each study group) as well as expression of HMGB1 and CD68 in donor and postreperfusion biopsy in 53 patients (Tac 25, control 28). The increment of IL-17 level between 1 and 0 time points was significantly lower in Tacrolimus group. Unlike to IL-17 the levels of IL-6, 8, 23, TNF-a, MIP-1a and P-selectin as well as expression of HMGB1 were comparable between groups. There was a trend toward lower postreperfusion CD68 increase in the Tacrolimus group. BACKGROUND It have been shown that tacrolimus (Tac) posess the activity to suppress inflammation and immune response in the transplanted liver on a genome-wide basis (Kristo I., Transpl Int.2011). Previous protocols used very small sample sizes and implemented either only portal Tac flushing (Kristo) or in vivo arterio- portal flushing (Shawn D. St. Peter., Liver Transpl. 2003). In our study we decided to expand the donor liver exposure to tacrolimus during back-table preparation. METHODS A prospective randomized study was conducted (ClinicalTrials.gov Identifier: NCT ). In the intervention group (38 pts) 1000 ml of cooled to 4C HTK solution containing Tacrolimus (20 ng/ml) was given at back-table through intraportal (gravity pressure 40 cm) and intraarterial infusion (pressure mmHg) followed by intraportal infusion of cooled 200 ml 5% solution of Albumin containing 20 ng/ml Tacrolimus (gravity pressure 40 cm). After infusuion liver was left resting in effluent until transplant. No tacrolimus was added in the control group (34 pts). Exclusion criteria: pediatrics under 18 y.o., split, reduced, ALF and multiorgan failure. Primary Outcome Measures: EAD based on recent criteria (Olthoff KM, Liver Transpl. 2010).Secondary Outcome Measures: postreperfusion hyperfibrinolysis diagnosed by Thromboelastometry at 15 min and 2h after portal reperfusion. PNF was defined as EAD leading to death or retransplantation. Study was restricted to classic technique of LTx with sequential portal-arterial reperfusion. Average data are shown as Median. Complimentary Luminex assay and immunohistochemistry were done to investigate the main cytokines and pathways involved. Blood sampling before donor organ procurement Liver biopsy during donor organ procurement Back-Table v.portae and hepatic artery flushing with tacrolimus or HTK only followed by albumin flushing with tacrolimus or no tacrolimus followed by resting in effluent until transplant Blood samples at liver transplant procedure Liver biopsy 2 hours postrepefusion Blood samples on day 1 and 3 [2,3] v.portae, just before unclamping IVC, just before unclamping Hepatic veins, after v.portae unclamping [0] Hepatic veins 40 min after portal reperfusion [1] IL-2R, IL-6, IL- 8,TLR4, Lps-Bp Not yet completed for TLR4 gene sequencing and staining for CD-68 and HMGB1 Partly completed Main (38)Control (34) Lipopolisaccarid binding protein, IL-6 Not yet completed P-selectin, IL-6,8,17,23, TNF-a, MIP-1a Partly completed staining for CD-68 and HMGB1 Partly completed IL-6,8,17,23 TNF-a, MIP-1a, VEGF, Elastase, Lps- Bp Not yet completed