High-dose methylprednisolone therapy in multiple sclerosis induces apoptosis in peripheral blood leukocytes
High-dose methylprednisolone therapy in multiple sclerosis induces apoptosis in peripheral blood leukocytes
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DOI：
10.1001/archneur.58.1.91
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发表时间：
2001-01-01
影响因子：
--
通讯作者：
Gold, R
中科院分区：
文献类型：
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作者：
Leussink, VI;Jung, S;Gold, R
背景：在多发性硬化症（MS）的自然病程中，细胞凋亡被认为有助于消除炎症中枢神经系统中的T细胞。在实验性自身免疫性脑脊髓炎动物模型中，高剂量糖皮质激素（GC）可诱导t细胞凋亡。目的：探讨静脉注射大剂量GC对MS患者体外t细胞凋亡的影响；66名多发性硬化症患者（28名复发缓解型多发性硬化症患者，22名继发性慢性进展性多发性硬化症患者，16名原发性慢性进展性多发性硬化症患者）和16名接受皮质类固醇治疗其他疾病的对照患者纳入研究。方法：于清晨2小时内首次给药500 ~ 1000 mg甲基强的松龙前后立即采血。梯度分离的外周血白细胞（PBLs）用皮质类固醇（阳性对照）、有丝分裂原植物血凝素或抗t细胞受体单克隆抗体培养，未经刺激。为了研究细胞凋亡，将pbl培养过夜，并通过TUNEL（终端转移酶介导的dUTP生物素nick末端标记）或膜联蛋白标记联合CD4、CD8、CD22、CD56或bcl-2染色，通过免疫流式细胞术进行分析。用h -3-胸腺嘧啶掺入法测定细胞增殖。培养48小时后收集上清液进行细胞因子测定。结果：经体内皮质类固醇治疗后，3个MS亚组中未受刺激的pbl细胞凋亡均明显增加。荧光活化细胞分选分析显示凋亡主要影响CD4 T细胞。GC治疗后自然杀伤细胞相对增加，但凋亡细胞率未见变化。高剂量GC治疗对t细胞亚群中bcl-2的表达无显著影响。与脉冲治疗前相比，GC治疗后t细胞受体刺激的pbl培养上清含有较低浓度的白细胞介素2、干扰素γ和肿瘤坏死因子α。在对照组中，细胞凋亡率和细胞因子的产生也发生了类似的变化。结论：皮质类固醇脉冲治疗是白细胞凋亡的强诱导剂。细胞凋亡的诱导可能有助于t细胞活性的下调，从而终止中枢神经系统的炎症。
Background: Apoptosis is supposed to contribute to the elimination of T cells fi om the inflamed central nervous system in the natural disease course of multiple sclerosis (MS). In the animal model experimental autoimmune encephalomyelitis, T-cell apoptosis can be induced by high-dose glucocorticoid (GC) administration.Objective: To study the effects of intravenous high-dose GC therapy in MS on T-cell apoptosis ex vivo.Patients; Sixty-six patients with MS (28 with relapsing-remitting MS, 22 with secondary chronic progressive MS, and 16 with primary chronic progressive MS) and 16 control patients receiving corticosteroids for other disorders were included in the study.Methods: Blood samples were collected before and immediately after the first infusion of 500 to 1000 mg of methylprednisolone given during 2 hours in the early morning. Gradient-isolated peripheral blood leukocytcs (PBLs) were cultured, unstimulated, with cortiro-steroids (positive control), the mitogen phytohemagglutinin, or anti-T-ccll receptor monoclonal antibody. For investigation of apoptosis, PBLs were cultured overnight and analyzed by immunoflow cytometry using TUNEL (terminal transferase-mediated dUTP biotin nick end labeling) or annexin labeling in combination with CD4, CD8, CD22, CD56, or bcl-2 staining. Proliferation was measured by H-3-thymidine incorporation. For cytokine determination, supernatants were collected after 48 hours of culture.Results: After in vivo corticosteroid treatment, apoptosis of unstimulated PBLs was markedly and significantly augmented in all 3 MS subgroups. Fluorescence-activated cell sorter analysis showed that apoptosis affected predominantly CD4 T cells. Natural killer cells showed a relative increase after GC therapy without a change in the rate of apoptotic cells. Expression of bcl-2 in T-cell subpopulations was not significantly modified by highdose GC therapy. Culture supernatants of T-cell receptor-stimulated PBLs after GC therapy contained lower concentrations of interleukin 2, interferon gamma, and tumor necrosis factor alpha than those from PBLs taken before pulse therapy. Similar changes in the rate of apoptosis and cytokine production were seen in controls.Conclusions: Corticosteroid pulse therapy is a strong inducer of leukocyte apoptosis. Induction of apoptosis might contribute to the down-regulation of T-cell activity and thereby terminate inflammation in the central nervous system.