Gas-generating systems in acute renal allograft rejection in the rat: Co-induction of heme oxygenase and nitric oxide synthase

Gases are now viewed as biologic messengers, and in this regard, carbon monoxide and nitric oxide are incriminated in signaling processes in neural tissue. Carbon monoxide is generated by heme oxygenase (HO), an enzyme inducible by heme, cytokines, and oxidative stress and considered an antioxidant response; nitric oxide is generated by nitric oxide synthase, an enzyme also inducible by cytokines. Since mononuclear cells infiltrate the acutely rejecting kidney, and foster within the kidney oxidative stress and a cytokine-enriched milieu, we examined the expression of these enzymes in acute renal allograft rejection (AR) (Brown Norway kidney to a Lewis rat; n=17) and in control isografts (Lewis kidney to a Lewis rat; n=17). No immunosuppressives were used. We found marked induction of HO mRNA and protein in renal allografts at day 5 after transplantation. Prominent expression of HO protein, as detected by immunofluorescence, was observed in the mononuclear cells infiltrating the renal allograft. More than 80% of these cells were macrophages, as identified by positive staining with ED1 antibody. ED1⁺ cells were rare in isografts and did not stain for HO. We also found co-expression of mRNA and protein for the inducible isoform of nitric oxide synthase (iNOS) in AR at day 5 after transplantation. Induction of HO and iNOS may reflect the cellular effect of diverse cytokines elaborated in the rejecting kidney. HO may enable the macrophage to degrade heme-containing proteins released from erythrocytes and other damaged cells; alternatively, induction of HO may defend the macrophage against oxidant injury. Increased nitric oxide, as a result of iNOS activity, may antagonize the vasoconstrictive effects of a number of mediators (i.e., thromboxane and endothelin) present in acute rejection; conversely, nitric oxide may prove cytotoxic through a number of recognized effects. Our studies provide the first demonstration of the induction of HO in the rejecting renal allograft as well as the first demonstration in vivo for the induction of HO in macrophages at the site of an inflammatory response. Such expression, linked as it is to the expression of iNOS, indicates that the macrophage mimics the behavior of neural cells by generating these gaseous messengers; thus, neural cells are not alone in deploying these mediators. Through a number of effects, these products of HO and iNOS may influence the nature and severity of tissue injury in AR.