OR17 Heme oxygenase-1 modulates HLA class I antibody-dependent endothelial cell activation

Abstract

Aim Antibody-mediated rejection (AMR) is a key limiting factor for long-term graft survival in heart and kidney transplantation. Activation of endothelial cells (ECs) via complement-independent effects of human leukocyte antigen class I (HLA I) antibodies (Abs) plays a major role in the pathogenesis of AMR. As the antioxidant enzyme heme oxygenase (HO)-1 is known to have cell type-specific anti-inflammatory effects in the endothelium, we investigated its role on HLA I Ab-dependent activation of human ECs. Methods Regulation of inducible proinflammatory endothelial adhesion molecules and chemokines (VCAM-1, ICAM-1, IL-8 and MCP-1) by monoclonal pan- and allele-specific HLA I Abs was determined in cell cultures of primary human umbilical venous, aortic macrovascular and microvascular ECs. HO-1 was modulated by pharmacological regulators and siRNA-mediated knockdown. Adherence of THP-1 monocytes to ECs was determined by leukocyte adhesion assay. Results Exposure of human macro- and microvascular EC cultures to HLA I Abs caused endothelial activation, as indicated by up-regulation of VCAM-1, ICAM-1, MCP-1 and IL-8. This up-regulation was mediated via the phosphatidylinositol-3 kinase (PI3K)/Akt and NF- κ B pathways. Pharmacological induction of HO-1 with cobalt-protoporphyrin IX reduced, whereas inhibition of HO-1 with either zinc-protoporphyrin IX or siRNA-mediated knockdown increased HLA I Ab-dependent EC activation. Binding of THP-1 monocytes was enhanced in HLA I Ab-stimulated ECs. This effect was counteracted by HO 1 up-regulation. Conclusion HLA I Ab-dependent EC activation is modulated by specific HO-1 up-regulation. Thus, targeted regulation of endothelial HO-1 may be a novel therapeutic approach for the treatment of AMR in kidney and heart transplantation. S. Immenschuh: Grant/Research Support; Company/Organization; Else Kroner-Fresenius Stiftung EKFS 2012_A309.