Jonsson C A, Carlsten H
Department of Rheumatology, Göteborg University, Sweden.
Clin Exp Immunol. 2001 Jun;124(3):486-91. doi: 10.1046/j.1365-2249.2001.01545.x.
The aim of this study was to evaluate the effects of the immunomodulating drug mycophenolic acid (MPA) on splenocytes in an animal model of systemic lupus erythematosus (SLE), using MRLlpr/lpr mice. MPA reversibly inhibits inosine 5'-monophosphate dehydrogenase, an enzyme involved in the de novo guanosine synthesis. Splenocytes were treated with MPA (at 1 or 10 microM), and stimulated with either lipopolysaccharide (LPS; 10 microg/ml) or concanavalin A (ConA; 1.25 microg/ml). In blocking experiments, guanosine (100 microM) was added to the cultures to inhibit the effects of MPA. Lymphocyte proliferation, enumeration of immunoglobulin producing cells (using ELISPOT) and quantification of anti-double-stranded (ds) DNA antibodies, IFN-gamma and IL-10 (by ELISA) in supernatants were performed. In addition, cell viability was evaluated using propidium iodide and flow cytometry. We found that MPA-treated splenocytes had dramatically decreased mitogen-induced proliferation and number of immunoglobulin producing cells, down-regulated production of IFN-gamma, IL-10 and IgM anti-dsDNA antibodies. The viability of MPA-treated cells was also decreased. All of the effect modulated by MPA could be neutralized by the addition of guanosine. We conclude that MPA has potent immunomodulating effects on both B and T lymphocytes, modulating not only proliferation, but also the production of cytokines, immunoglobulins and autoantibodies.
本研究旨在使用MRLlpr/lpr小鼠,评估免疫调节药物霉酚酸(MPA)对系统性红斑狼疮(SLE)动物模型脾细胞的影响。MPA可逆性抑制肌苷5'-单磷酸脱氢酶,该酶参与鸟苷的从头合成。脾细胞用MPA(1或10 microM)处理,并用脂多糖(LPS;10 microg/ml)或伴刀豆球蛋白A(ConA;1.25 microg/ml)刺激。在阻断实验中,向培养物中加入鸟苷(100 microM)以抑制MPA的作用。进行淋巴细胞增殖、免疫球蛋白产生细胞计数(使用ELISPOT)以及上清液中抗双链(ds)DNA抗体、IFN-γ和IL-10的定量(通过ELISA)。此外,使用碘化丙啶和流式细胞术评估细胞活力。我们发现,MPA处理的脾细胞有丝分裂原诱导的增殖和免疫球蛋白产生细胞数量显著减少,IFN-γ、IL-10和IgM抗dsDNA抗体的产生下调。MPA处理细胞的活力也降低。MPA调节的所有效应均可通过添加鸟苷来中和。我们得出结论,MPA对B淋巴细胞和T淋巴细胞均有强大的免疫调节作用,不仅调节增殖,还调节细胞因子、免疫球蛋白和自身抗体的产生。