基因治疗人类多发性硬化症的可能性研究:腺病毒转染CTLA-4Ig对动物实验性自身免疫性脑脊髓炎病理过程的抑制或阻断作用(英文)

Possibility of gene therapy in the treatment of human multiple sclerosis:the inhibition or blocking effects of adenovirus-transfected CTLA-4Ig on the pathologic process of experimental autoimmune encephalomyelitis animals

背景:实验性自身免疫性脑脊髓炎(experimentalautoimmuneen-cephalomyelitis,EAE)同人类的多发性硬化症(multiplesclerosis,MS)的病理机制相类似,因此认识和阻断EAE的病理过程是攻克MS的关键。目的:寻求一种可能抑制或阻断EAE病理过程,加速神经功能恢复进程的方法。设计:随机对照的实验研究。单位:解放军第三军医大学附属西南医院神经内科和烧伤研究所。材料:实验在解放军第三军医大学动物实验室完成。实验材料:豚鼠4只(雌雄不限,体质量300～400g),Wistar大鼠40只(雌雄不限,体质量180～200g)。方法:用所构建的CTLA-4Ig腺病毒通过侧脑室注射到Wistar大鼠EAE模型体内观察其疗效,用冷冻切片观察CTLA-4Ig在脑组织中的表达,观察发病情况、病理改变。主要观察指标:①两组大鼠致敏后的发病天数、发病只数及潜伏期及神经功能评分情况。②光镜下观察两组大鼠病理学改变情况。结果:发病情况:生理盐水对照组大鼠于致敏后第10天开始发病。于第16天全部发病,平均发病天数为12.2d,临床评分平均为3.1;腺病毒介导的CTLA-4Ig组于致敏后第14天开始发病,第18天共发病6只,平均发病天数为14.1d,临床评分平均为2.4。病理结果:侧脑室注射AdCTLA-4Ig后24,48,72h取脑组织做冷冻切片在荧光显微镜下观察侧脑室周围和脊髓有荧光。在?

BACKGROUND:The pathogenesis of experimental autoimmune encephalomyelitis(EAE)is similar to human multiple sclerosis(MS),hence, the comprehension and interdiction of EAE pathologic process is the key to capture MS. OBJECTIVE:To find a possible method to inhibit or interdict EAE pathologic process to speed up the restorative course for neural function. DESIGN:A randomized controlled trial SETTING:Department of Neurology and Institute for Burn of Southwest Hospital Affiliated of Third Military Medical University of Chinese PLA MATERIALS:The study was performed in the laboratory for experimental animal of the Third Military Medical University of Chinese PLA.Four guinea-pigs in either gender with a mass from 300 g to 400 g and 40 Wistar rats in either gender with a mass from 180 g to 200 g were included in the study. INTERVENTIONS:The constructed CTLA-4Ig adenovirus were injected into the lateral cerebral ventricle of EAE model Wistar rats to observe its therapeutic effectiveness.The expression of CLTA-4Ig in the brain tissue was observed in frozen sections.The morbidity of the disease and the pathologic changes were observed as well. MAIN OUTCOME MEASURES:①attack days,numbers and latency in rats of two groups after sensitization, and the evaluation of neural function;②pathologic changes in rats of two groups under optical microscope RESULTS:Morbidity:Rats of saline group started to be attacked on the 10th day after sensitization and all of them were suffered on the 16th day,with an average latency of 12.2 days and mean clinical evaluation score of 3.1 point.Rats of adenovirus-mediated CTLA-4Ig group began to be attacked on the 14th day after sensitisation and a total of 6 rats were attacked on the 18th day with an average latency of 14.1 days and mean clinical evaluation score of 2.4 point.Pathologic results:The brain tissues were harvested on the 24 hours,48 hours and 72 hours after adenovirus-mediated CTLA-4Ig injection into the lateral cerebral ventricle for frozen sectioning,which were observed under fluorescence microscope.Fluorescence could be seen at entourage of the lateral cerebral ventricle and spine.Under optical microscope in control group,lots of infiltrations of inflammatory cells in the brain and spine could be seen,of which inflammatory cells aggregated around vessels with a sleevelet change.Myelin staining(Loyez staining)showed demyelination. In adenovirus-mediated CTLA-4Ig group,the infiltration of inflammatory cells in brain and spine significantly reduced,and the formation of vascular sleevelet change and demyelination reduced as well. CONCLUSION:CTLA-4Ig can induce the deactivation of T cell,which can effectively inhibit cellular and humoral immunity.It can interdict the pathologic process of EAE,an autoimmune disease mainly due to the excessive activation of T cells to enhance the neural function score of the rats and effectively improve the neural functional symptoms.