Objective. Inhibition of T cell DNA methylation causes autoreactivity in vitro and a lupus-like disease in vivo, suggesting that T cell DNA hypomethylation may contribute to autoimmunity. The hypomethylation effects are due, in part, to overexpression of lymphocyte function-associated antigen 1 (LFA-1) (CD11a/CD18). Importantly, T cells from patients with active lupus have hypomethylated DNA and overexpress LFA-1 on an autoreactive subset, suggesting that the same mechanism could contribute to human lupus. The present study investigated the nature of the methylation change that affects LFA-1 expression in vitro and in human lupus.Methods. Bisulfite sequencing was used to determine the methylation status of the ITGAL promoter and flanking regions in T cells from lupus patients and healthy subjects, and in T cells treated with DNA methylation inhibitors. "Patch" methylation of promoter sequences in reporter constructs was used to determine the functional significance of the methylation changes.Results. Hypomethylation of specific sequences flanking the ITGAL promoter was seen in T cells from patients with active lupus and in T cells treated with 5-azacytidine and procainamide. Patch methylation of this region suppressed ITGAL promoter function.Conclusion. DNA methylation changes occur in specific sequences that regulate LFA-1 expression in lupus T cells and in the hypomethylation model, indicating that altered methylation of specific genes may play a role in the pathogenesis of lupus.The mechanisms initiating human systemic lupus erythematosus (SLE) remain unknown. The finding that exposure to certain drugs can induce a lupus-like disease has provided leads into the nature of biochemical alterations associated with lupus. The 2 drugs most frequently associated with lupus, procainamide and hydralazine, can cause a lupus-like disease through effects on T cell DNA methylation. Treating T cells with procainamide, hydralazine, or 5-azacytidine (5-azaC; the prototypic DNA methylation inhibitor) demethylates DNA, alters gene expression, and induces major histocompatibility complex-specific T cell autoreactivity (1-5). Adoptive transfer of the autoreactive cells causes a lupus-like disease in animal models (5-7). The autoimmune effects of the methylation inhibitors are due, in part, to overexpression of lymphocyte functionassociated antigen 1 (LFA-1) (CD11a/CD18), because increasing T cell LFA-1 by transfection causes an identical autoreactivity in vitro, and a similar autoimmune disease in vivo (4,8).Altered DNA methylation has also been implicated in human lupus. T cells from patients with active lupus have an ϳ17% decrease in genomic deoxymethylcytosine content and overexpress LFA-1 on an autoreactive subset (9-11). However, whether the DNA hypomethylation occurring in lupus affects transcriptionally relevant regions is not known. It is similarly unknown if the DNA hypomethylation induced by 5-azaC or procainamide affects the same sequences as those affected in SLE.DNA methylation inhibitors increase LFA-1 through thei...

Background: Kawasaki disease produces a vasculitis which may injure the coronary arteries and predispose these patients to early atherosclerosis. Previously, case reports have documented that Kawasaki patients may develop coronary calcifications and may die of sudden death many years after their acute illness. Ultrafast computer tomography (UCT) has been successful at detecting coronary calcifications in adults with atherosclerosis. The goal of this study is to show that UCT can be utilized as a non-invasive technique to identify Kawasaki patients at high risk for coronary disease and who would require long-term surveillance through adulthood.Methods and Results: Eighteen patients ages five to twenty-one years old with a history of Kawasaki disease were enrolled. Each patient had an UCT of the heart. There were four patients with calcifications noted. All of the four patients had: a previous history of an aneurysm, were at least five years from their acute illness, and had the calcification correlate to the previous site of aneursym formation. A calcium score was determined by the Agaston method for each calcification. The Mayo clinic guidelines for adult atherosclerosis were then used to correlate these scores with the risk of future coronary artery disease. All four patients were at moderate or greater risk of future coronary disease.Conclusion: This study indicates that UCT can be used as an effective non-invasive methodology in Kawasaki patients to successfully identify the high-risk patients predisposed to developing potential early atherosclerotic coronary artery disease.