A major manifestation of chronic allograft failure (CAF) is the accelerated onset of atherosclerotic lesions within the graft. Polymorphisms in the endothelial nitric oxide synthase (eNOS) gene have been implicated in the pathogenesis of native atherosclerosis. This study tested the hypothesis that polymorphisms in eNOS are associated with susceptibility to CAF after cadaveric renal transplantation. The patient cohort comprised 140 renal transplant recipients who had received their transplants between 1985 and 1997 at the Oxford Transplant Centre and included 61 patients with biopsy-proven CAF and 79 with stable graft function for at least 10 years (long-term survivors, LTS). Genotyping for one polymorphism in the promoter region and two polymorphisms in the coding regions of the eNOS gene was performed by polymerase chain reaction with sequence-specific primers (PCR-SSP). No association was found between any genetic variant and the development of CAF, even after stratification for other known risk factors. Statistical analysis revealed that all three polymorphisms were closely linked. We conclude that recipient eNOS gene polymorphisms do not alter the risk of CAF after renal transplantation.