干扰素-β治疗实验性自身免疫性脑脊髓炎可导致T细胞浸润迅速非凋亡性终止。

Interferon-beta treatment of experimental autoimmune encephalomyelitis leads to rapid nonapoptotic termination of T cell infiltration.

作者信息

Schmidt J, Stürzebecher S, Toyka K V, Gold R

机构信息

Department of Neurology, Clinical Research Group for Multiple Sclerosis and Neuroimmunology, University of Würzburg, Würzburg, Germany.

出版信息

J Neurosci Res. 2001 Jul 1;65(1):59-67. doi: 10.1002/jnr.1128.

DOI:10.1002/jnr.1128

PMID:11433430

Abstract

We investigated the possible mechanisms how interferon (IFN)-beta may control T cell infiltration in the CNS in experimental autoimmune encephalomyelitis (EAE). Adoptive transfer (AT) EAE was induced in groups of six female Lewis rats. Animals were treated with 3 x 10(5) units of recombinant rat IFN-beta s.c. once at 18 hr, or with 10 mg/kg methylprednisolone (MP) i.v. twice at 18 and 6 hr prior to dissection, or with a combination of both. T cell apoptosis was detected by immunohistochemistry on paraffin sections of spinal cord, using morphological criteria and TUNEL staining. Double labeling of immune cells was done for tumor necrosis factor (TNF)-alpha and metalloproteinase (MMP) 2. Disruption of the blood-brain barrier (BBB) was visualized by staining for albumin. In severe EAE, an increase of T cell apoptosis was seen after IFN-beta alone (all data presented as mean +/- SD: 24.5% +/- 2.2%, P < 0.05, vs. 19.4% +/- 3.1% in controls), and in combination with MP (29.4% +/- 7.3%, P < 0.05 vs. controls). Only the combination therapy decreased T cell infiltration (53.9 +/- 17.7 cells/mm(2), P < 0.05, vs. 99.5 +/- 35.2 cells/mm2 in controls). In moderate EAE, the rate of T cell apoptosis was slightly increased after IFN-beta (21.2% +/- 5.2% vs. 17.4% +/- 5.0% in controls), whereas MP alone (25.5% +/- 3.5%, P < 0.01 vs. controls) and the combination therapy (22.4% +/- 4.8%, P < 0.05 vs. controls) had a clear augmenting effect. IFN-beta tended to decrease T cell infiltration (46.1 +/- 12.7 cells/mm2) compared to controls (59.2 +/- 18.5 cells/mm2). The rate of TNF-alpha-expressing T cells was significantly decreased by IFN-beta and in combination with MP. Also, TNF-alpha expression in macrophages was significantly reduced by IFN-beta and by the combination therapy. The rate of MMP2-expressing macrophages was lower after IFN-beta but clearly decreased only in combination with MP. BBB disruption was ameliorated after IFN-beta but significantly only in combination with MP. Our study indicates that IFN-beta affects the immunopathological process in EAE in several ways, but apoptosis appears as a minor component. In view of treatment of MS relapses, the synergistic effects in this study corroborate the use of a combination therapy with high-dose MP and IFN-beta.

摘要

我们研究了干扰素（IFN）-β在实验性自身免疫性脑脊髓炎（EAE）中控制中枢神经系统（CNS）T细胞浸润的可能机制。将六只雌性Lewis大鼠分为几组，通过过继转移（AT）诱导EAE。在解剖前18小时，给动物皮下注射3×10⁵单位重组大鼠IFN-β一次，或静脉注射10mg/kg甲泼尼龙（MP）两次，分别在解剖前18小时和6小时，或两者联合使用。通过对脊髓石蜡切片进行免疫组织化学，利用形态学标准和TUNEL染色检测T细胞凋亡。对免疫细胞进行肿瘤坏死因子（TNF）-α和金属蛋白酶（MMP）2的双重标记。通过白蛋白染色观察血脑屏障（BBB）的破坏情况。在严重EAE中，单独使用IFN-β后可见T细胞凋亡增加（所有数据均以平均值±标准差表示：24.5%±2.2%，P<0.05，对照组为19.4%±3.1%），联合MP后（29.4%±7.3%，P<0.05 vs.对照组）也是如此。只有联合治疗降低了T细胞浸润（53.9±17.7个细胞/mm²，P<0.05，对照组为99.5±35.2个细胞/mm²）。在中度EAE中，IFN-β后T细胞凋亡率略有增加（21.2%±5.2%，对照组为17.4%±5.0%），而单独使用MP（25.5%±3.5%，P<0.01 vs.对照组）和联合治疗（22.4%±4.8%，P<0.05 vs.对照组）有明显的增强作用。与对照组相比，IFN-β倾向于降低T细胞浸润（46.1±12.7个细胞/mm²）（对照组为59.2±18.5个细胞/mm²）。IFN-β单独使用以及与MP联合使用均显著降低了表达TNF-α的T细胞比例。此外，IFN-β和联合治疗均显著降低了巨噬细胞中TNF-α的表达。IFN-β后表达MMP2的巨噬细胞比例较低，但仅在与MP联合使用时明显降低。IFN-β后BBB破坏有所改善，但仅在与MP联合使用时显著改善。我们的研究表明，IFN-β以多种方式影响EAE的免疫病理过程，但凋亡似乎是次要成分。鉴于MS复发的治疗，本研究中的协同效应证实了高剂量MP和IFN-β联合治疗的应用。