The effect of allo-injury in an in vitro model of allograft microvasculature

Endothelial cells are critical to the integrity of allograft vasculature and can be damaged by alloreactive T cells or soluble mediators of alloreactivity. The biochemical effects of T cell-mediated damage to the endothelial cells have been studied, but not the structural and morphological effects of alloinjury on endothelial cells in the allograft. We utilized an assay that reproduces microvasculature in vitro to study the effect of alloreactivity on endothelial cells. In this assay, endothelial cells are induced into capillary-like networks that simulate microvascular capillaries. We studied the effect of allogeneic T cells and of soluble mediators from both mixed lymphocyte cultures (MLCs) and rejected heart allograft tissue on the in vitro capillaries. We found that both allogeneic T cells and soluble mediators inhibit the formation of the in vitro endothelial capillaries, suggesting that they cause a mild-to-moderate dysfunction of the endothelial cells. The inhibitory effect of the soluble mediators seems to be mediated, at least partly, by IFN-γ, since this effect was prevented by antibody to IFN-γ. Furthermore, pre-incubation of the in vitro capillaries with IFN-γ appeared to magnify the effect of allogeneic T cells, as shown by a complete breakdown of well-formed in vitro capillary networks. Our experiments suggest that the in vitro capillary-tube model reflects structural injury to allograft vasculature by alloreactive T cells and their soluble mediators.