THE RISK OF EARLY LIVER ALLOGRAFT DYSFUNCTION IS ASSOCIATED WITH THE TLR-4 GENE GENOTYPE IN THE RS913930 SEQUENCE AND IS IMPLEMENTED VIA HMGB1 NUCLEAR PROTEIN, KUPFFER CELLS AND IL-23 ACTIVATION

Aim. To evaluate the associations of genotypes of clinically relevant nucleotides rs11536865, rs913930 and rs5030717 of the TLR-4 gene with the risk of development and severity of early allograft dysfunction after liver transplantation. Materials and methods. A case-control study enrolling 71 patients was organized. Inclusion criteria: DBD liver transplantation. Exclusion criteria: living related liver transplantation, reduced graft transplantation, recipient’s age fewer than 18. Results. Within rs5030717 there were identifi ed three genotypes: AA (81.6%) and two genotypes with the minor G-allele: AG (12.6%) and GG (5.6%). Within rs913930 there identi- fi ed three genotypes: TT (59.1%) and two genotypes with the minor C-allele: C/T (29.5%) and CC (11.2%). The rs11536865 studying revealed no polymorphism (GG genotype). The early allograft liver dysfunction (EAD) developed in 19.7% of patients, the severe EAD in 11.2% of patients, septic complications in 14%, acute cellular rejection in 23.9% of cases. The C/T genotype of the TLR-4 gene in the SNP rs913930 sequence was closely associated with the EAD development (OR 4.8 to 1; p = 0.047; 95% CI 1–23.4). Рatients with the donor’s liver C/T genotype had a reliably higher proportion (%) of the HMGB1 positive hepatocytes in the donor’s bioptate, 21 (17–29%) vs the СС+TT genotypes, 16 (10–19%) (Mann–Whitney test, p = 0.01). The CD68 expression in the liver bioptate at the donor’s stage was reliably higher in the carriers of heterozygotes in the SNP rs913930 (C/T genotype) and in the SNP rs5030717 (AG genotype), (Mann–Whitney test, p = 0.03). Signifi cant positive correlation between the CD68 expression in the donor’s liver bioptates and the IL-23 level in the hepatic vein has been determined in an hour after the portal reperfusion (ρ = 0.62; p = 0.04) as well as between the HMGB1 expression in the donor’s liver bioptates and the АSТ level in 24 hours after the reperfusion (r = 0.4; p = 0.02). The HMGB1 staining in the donor’s liver bioptates was higher in the EAD patients, 21 (20; 29) cells/mm2 in comparison with the patients without EAD, 16 (12; 18) (Mann–Whitney test, p = 0.0036). Conclusion. The early allograft liver dysfunction is associated with the genetic predisposition caused by the TLR-4 gene polymorphism and is implemented via the HMGB1, Kupffer cells and IL-23 activation. 

1. Deschenes M. Early allograft dysfunction: Causes, recognition, and management. Special Issue: Liver Transplan ta tion. 2013 Nov; 19 (S2): S6–S8.

2. Bezinover D, Kadry Z, McCullough P, McQuillan PM, Uemura T, Welker K et al. Release of cytokines and hemodynamic instability during the reperfusion of a liver graft. Liver Transplantation. 2011 Mar; 17 (3): 324–330.

3. Conti A, Scala S, D’Agostino P, Alimenti E, Morelli D, Andria B et al. Wide Gene Expression Profi ling of Ischemia-Reperfusion Injury in Human Liver Transplantation. Liver Transplantation. 2007; 13: 99–113.

4. Wang X, Sun R, Wei H, Tian Z. High-Mobility Group Box 1 (HMGB1)-Toll-Like Receptor (TLR)4-Interleukin (IL)-23-IL-17A Axis in Drug-Induced Damage-Associated Lethal Hepatitis: Interaction of γδ-T Cells with Macrophages. Hepatology. 2013; 57: 373–384.

5. Howell J, Gow P, Angus P, Visvanathan K. Role of toll-like receptors in liver transplantation. Liver Transplantation. 2014 Mar; 20 (3): 270–280.

6. Shen XD, Ke B, Zhai Y, Gao F, Tsuchihashi S, Lassman CR et al. Absence of toll-like receptor 4 (TLR4) signaling in the donor organ reduces ischemia and reperfusion injury in a murine liver transplantation model. Liver Transplantation. 2007 Oct; 13 (10): 1435–4314.

7. Ellett JD, Atkinson C, Evans ZP, Amani Z, Balish E, Schmidt MG et al. Toll-like receptor 4 knockout mice are protected from endothelial overactivation in the absence of Kupffer cells after total hepatic ischemia/reperfusion. Liver Transplantation. 2011; 17 (9): 1089–1098.

8. Ellett JD, Evans ZP, Atkinson C, Schmidt MG, Schnellmann RG, Chavin KD. Toll-Like Receptor 4 is a Key Mediator of Murine Steatotic Liver Warm Ischemia/ Reperfusion Injury. Liver Transplantation. 2009; 15 (9): 1101–1109.

9. Oetting WS, Guan W, Schladt DP, Leduc RE, Jacobson PA, Matas AJ et al. Donor Polymorphisms of Toll-Like Receptor 4 Associated With Graft Failure in Liver Transplant Recipients. Liver Transplantation. 2012; 18: 1399–1405.

10. http://www.ncbi.nlm.nih.gov/snp

11. Olthoff KM, Kulik L, Samstein B, Kaminski M, Abecassis M, Emond J et al. Validation of a current defi nition of early allograft dysfunction in liver transplant recipients and analysis of risk factors. Liver Transplantation. 2010; 16: 943–949.

12. Salvalaggio PR, Felga GE, Afonso RC, Ferraz-Neto BH. Early Allograft Dysfunction and Liver Transplant Outcomes: A Single Center Retrospective Study. Transplantation Proceedings. 2012; 44: 2449–2451.