N-Octanoyl-Dopamine inhibits cytokine production in activated T-cells and diminishes MHC-class-II expression as well as adhesion molecules in IFN\u03b3-stimulated endothelial cells

Björn B Hofmann,Matthias Kolibabka,Marloes Sol,Jana D Braun,Anna-Isabelle Kälsch,Bernhard K Krämer,Prama Pallavi,Yingchun Li,Carolina De La Torre,Nicolas Krapp,Benito A Yard

doi:10.1038/s41598-019-55983-1

Abstract

IFNγ enhances allograft immunogenicity and facilitates T-cell mediated rejection. This may cause interstitial fibrosis and tubular atrophy (IFTA), contributing to chronic allograft loss. We assessed if inhibition of T-cell activation by N-octanoyl dopamine (NOD) impairs adherence of activated T-cells to endothelial cells and the ability of activated T-cells to produce IFNγ. We also assessed if NOD affects IFNγ mediated gene expression in endothelial cells. The presence of NOD during T-cell activation significantly blunted their adhesion to unstimulated and cytokine stimulated HUVEC. Supernatants of these T-cells displayed significantly lower concentrations of TNFα and IFNγ and were less capable to facilitate T-cell adhesion. In the presence of NOD VLA-4 (CD49d/CD29) and LFA-1 (CD11a/CD18) expression on T-cells was reduced. NOD treatment of IFNγ stimulated HUVEC reduced the expression of MHC class II transactivator (CIITA), of MHC class II and its associated invariant chain CD74. Since IFTA is associated with T-cell mediated rejection and IFNγ to a large extent regulates immunogenicity of allografts, our current data suggest a potential clinical use of NOD in the treatment of transplant recipients. Further in vivo studies are warranted to confirm these in vitro findings and to assess the benefit of NOD on IFTA in clinically relevant models.

Highlights

Improvements in donor management[1,2], the continuous progress in immunosuppressive and supportive therapy[3] as well as refinements in organ preservation[4,5] have significantly contributed to better transplantation outcomes in the last decades
While no differences in T-cell adhesion were observed between both types of supernatants in a 1 to 2 dilution, T-cell adhesion decreased to the level of the control culture medium when higher dilutions of supernatants from T-cells that were activated in the presence of N-octanoyl dopamine (NOD) were used (Dilution 1/64 mean relative fluorescence units (RFU) ± SD: 40,180 ± 2,031 vs. 28,149 ± 1,462 vs. 26,812 ± 3,009; −NOD vs. +NOD vs. medium control p < 0.01) (Fig. 1B)
In keeping with the observation that T-cell adhesion to endothelial cells is diminished when T-cells are activated in the presence of NOD, we addressed if this was reflected by a reduced expression of T-cell ligands for adhesion molecules

Summary

Introduction

Improvements in donor management[1,2], the continuous progress in immunosuppressive and supportive therapy[3] as well as refinements in organ preservation[4,5] have significantly contributed to better transplantation outcomes in the last decades. Amongst the factors that influence allograft immunogenicity, interferon gamma (IFNγ) plays a prominent role It up-regulates the expression of MHC class I on hematopoetic and non hematopoetic cells and influences antigen presentation dendritic cells by formation of the so-called immune-proteasome (IP)[19,20]. In keeping with the pivotal role of IFNγ in regulating graft immunogenicity, the present study was conducted to assess if addition of NOD during T-cell activation impairs T-cell adhesion to unstimulated and IFNγ - or TNFα stimulated endothelial cells. We sought to address if supernatants of such activated T-cells are able to induce adhesion molecules on endothelial cells and if this is paralleled by the presence of IFNγ and TNFα in the supernatants. We addressed to what extent NOD influences IFNγ mediated gene expression in endothelial cells

Methods

Results

Conclusion