Steinman’s work was supported by grants from the NIH (R01-AI-59709) and the National Multiple Sclerosis Society. cytokine levels in vivo, were measured by ELISA. T cell proliferation was assessed by thymidine incorporation assay. Serum cholesterol levels were determined using a standard fluorometric assay. Kidney tissue was harvested and evaluated for pathologic changes. Results In NZB/NZW mice, oral atorvastatin had significant effects on T cell proliferation and cytokine production in vitro. Atorvastatin also induced significant increases in serum levels of interleukin-4. However, atorvastatin treatment in NZB/NZW mice had no significant impact on proteinuria, survival, serum anti-dsDNA antibody and cholesterol levels, or extent of renal disease. Conclusion Monotherapy with oral atorvastatin has no protective effects in a murine model of spontaneous SLE. The efficacy of atorvastatin in human SLE remains SSTR5 antagonist 2 TFA to be decided. Systemic lupus erythematosus (SLE) is usually a chronic autoimmune disease that affects more than 1 million people in the US. The clinically heterogeneous nature of SLE renders therapeutic intervention particularly challenging. Conventional approaches include the use of corticosteroids, cytotoxic brokers such as cyclophosphamide and azathioprine, and, more recently, investigational brokers such as mycophenolate mofetil and rituximab. These agents have been used with varying success and are associated with a considerable number of side effects (1C3). In addition to the myriad of clinical and laboratory manifestations that comprise the diagnostic criteria for SLE and related connective tissue diseases, it has become increasingly clear that patients with SLE have increased morbidity and mortality resulting from accelerated atherosclerosis (4). This realization has led to efforts by clinicians to reduce corticosteroid use, as well as to recommend patient lifestyle modifications. Statins, or 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase inhibitors, are cholesterol-lowering drugs that have displayed immunomodulatory effects in numerous models of inflammatory and autoimmune disease, including multiple sclerosis (MS) and rheumatoid arthritis. Although side effects have been reported, statins are generally well tolerated in SSTR5 antagonist 2 TFA humans, making these brokers ideal for potential use as therapeutic brokers in immune-mediated diseases. In this study, our goal was to determine whether monotherapy with atorvastatin could suppress clinical disease in the (NZB NZW)F1 (NZB/NZW) mouse model of spontaneous SLE. Materials and Methods Mice Female NZB/NZW mice were purchased from The Jackson Laboratory (Bar Harbor, ME) and maintained under standard conditions in the Research Animal Facility at SSTR5 antagonist 2 TFA Stanford University. Experiments were conducted in accordance with approved Institutional Animal Care and Use Committee and National Institutes of Health (NIH) guidelines. Reagents Atorvastatin (prescription formulation; Pfizer, New York, NY) was brought into suspension in phosphate buffered saline (PBS) at 0.4 mg/ml, and a 0.5-ml volume (equivalent to 10 mg/kg) was administered via oral gavage, once daily, using 20-mm feeding needles (Popper & Sons, New Hyde Park, NY). PBS was administered as a vehicle control. Treatment of the mice (n = 15) began at 20 weeks of age, and the surviving mice were killed at 40 weeks of age. Renal histopathologic analysis Proteinuria was assessed once weekly using Albustix reagent strips for urinalysis (Bayer, Elkhart, IN). At the end of the experiment, kidneys were harvested from surviving mice and fixed in 10% buffered formalin. Periodic acid-Schiff (PAS) staining was performed on paraffin-embedded sections, and kidney damage was scored according to standard NIH activity and chronicity indices (5) in a blinded manner by one of the authors (JPH). T cell proliferation assays Splenocytes were negatively selected for T cells via an enrichment column (R&D Systems, Minneapolis, MN). T cells ( 95% purity, as determined by flow cytometric analysis [data not Pten shown]) were cultured for 48 hours at 105 cells per well in 96-well Nunc plates (Nalgene, Rochester, NY) coated with anti-CD3 monoclonal antibody (2C11; BD PharMingen, San Diego, CA) and anti-CD28 monoclonal antibody (37.51; BD PharMingen), each at a concentration of 5 g/ml. Culture medium consisted of RPMI 1640 (Gibco, Carlsbad, CA) supplemented with L-glutamine (2 m 0.01; ***.