Lin En-Ju D, Symes C Wymond, Townsend-Nicholson Andrea, Klugmann Matthias, Klugmann Claudia B, Lehnert Klaus, Fong Dahna, Young Deborah, During Matthew J
Functional Genomics and Translational Neuroscience Lab, Department of Molecular Medicine and Pathology, University of Auckland 85 Park Road, Grafton, Auckland 1142, New Zealand ; Department of Molecular Virology, Immunology and Medical Genetics, The Ohio State University, BRT 912, 460 W 12th Avenue, Columbus, OH 43210, USA.
Functional Genomics and Translational Neuroscience Lab, Department of Molecular Medicine and Pathology, University of Auckland 85 Park Road, Grafton, Auckland 1142, New Zealand.
ISRN Neurosci. 2014 Apr 24;2014:103213. doi: 10.1155/2014/103213. eCollection 2014.
We have previously demonstrated the therapeutic potential of inducing a humoral response with autoantibodies to the N-methyl D-aspartate (NMDA) receptor using a genetic approach. In this study, we generated three recombinant proteins to different functional domains of the NMDA receptor, which is implicated in mediating brain tolerance, specifically NR1[21-375], NR1[313-619], NR1[654-800], and an intracellular scaffolding protein, Homer1a, with a similar anatomical expression pattern. All peptides showed similar antigenicity and antibody titers following systemic vaccination, and all animals thrived. Two months following vaccination, rats were administered the potent neurotoxin, kainic acid. NR1[21-375] animals showed an antiepileptic phenotype but no neuroprotection. Remarkably, despite ineffective antiepileptic activity, 6 of 7 seizing NR1[654-800] rats showed absolutely no injury with only minimal changes in the remaining animal, whereas the majority of persistently seizing rats in the other groups showed moderate to severe hippocampal injury. CREB, BDNF, and HSP70, proteins associated with preconditioning, were selectively upregulated in the hippocampus of NR1[654-800] animals, consistent with the observed neuroprotective phenotype. These results identify NR1 epitopes important in conferring anticonvulsive and neuroprotective effects and support the concept of an immunological strategy to induce a chronic state of tolerance in the brain.
我们之前已经证明了使用基因方法诱导针对N-甲基-D-天冬氨酸（NMDA）受体的自身抗体产生体液反应的治疗潜力。在本研究中，我们针对NMDA受体的不同功能域生成了三种重组蛋白，该受体与介导脑耐受性有关，具体为NR1[21-375]、NR1[313-619]、NR1[654-800]，以及一种具有相似解剖学表达模式的细胞内支架蛋白Homer1a。全身接种疫苗后，所有肽均表现出相似的抗原性和抗体滴度，且所有动物均茁壮成长。接种疫苗两个月后，给大鼠施用强效神经毒素海人酸。NR1[21-375]组动物表现出抗癫痫表型，但无神经保护作用。值得注意的是，尽管抗癫痫活性无效，但7只发作的NR1[654-800]大鼠中有6只完全没有损伤，其余动物仅有轻微变化，而其他组中大多数持续发作的大鼠表现出中度至重度海马损伤。与预处理相关的蛋白CREB、BDNF和HSP70在NR1[654-800]组动物的海马中选择性上调，这与观察到的神经保护表型一致。这些结果确定了在赋予抗惊厥和神经保护作用中重要的NR1表位，并支持了诱导大脑慢性耐受状态的免疫策略概念。
