Author:
Date:
2017-07-05
[Abstract] Experimental autoimmune encephalomyelitis (EAE) is an animal model of multiple sclerosis (MS), which is a chronic inflammatory disease of the central nervous system (CNS). It is characterized by focal demyelination and inflammatory responses mediated by myelin-specific autoreactive CD4+ T cells. Using a passive transfer model of EAE in mice, we have demonstrated that regional specific neural signals by sensory-sympathetic communications create gateways for immune cells at specific blood vessels of the CNS, a phenomenon known as the gateway reflex (Arima et al., 2012; Tracey, 2012; Arima et al., 2013; Sabharwal et al., 2014; Arima et al., 2015b). Here we describe protocols for passive transfer model of EAE using freshly isolated (MOG)-specific ...
[摘要] 实验性自身免疫性脑脊髓炎(EAE)是多发性硬化症(MS)的动物模型,其是中枢神经系统(CNS)的慢性炎性疾病。其特征在于由髓磷脂特异性自身反应性CD4 + T细胞介导的局灶性脱髓鞘和炎症反应。在小鼠中使用EAE的被动转移模型,我们已经证明,通过感觉交感通信的区域特异性神经信号在CNS的特定血管上产生免疫细胞的网关,这被称为网关反射(Arima等, ,2012; Tracey,2012; Arima等人,2013; Sabharwal等人,2014; Arima等人。 >,2015b)。在这里,我们描述了使用新鲜分离的(MOG)特异性CD4 + T细胞或周期性再刺激的MOG特异性CD4 + T细胞系的EAE的被动转移模型的方案,其是适用于体内追踪致病性CD4 T细胞,特别是CNS(Ogura等人,2008; Arima等人) 。,2012和2015b)。 【背景】广泛接受的是,自身反应性CD4 + T细胞在MS和EAE的发病机理中起重要作用(Reboldi,2009; International Multiple Sclerosis Genetics,et al。,2011; Steinman,2014),它们是CNS的慢性炎性疾病。 ...
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Author:
Date:
2015-10-20
[Abstract] Prominent accumulation of D-leucine, D-allo-isoleucine and D-valine was observed in the culture medium of the heterofermentative bacterial species, Lactobacillus otakiensis (L. otakiensis) JCM 15040. The racemase enzyme that resulted in this accumulation, isoleucine 2-epimerase, was purified from the bacterial cells. This is the first reported observation of such production of D-branched chain amino acids in lactic acid bacteria, and the first example of a racemase with isoleucine 2-epimerase activity in any organisms. In the described protocol, we introduce methods for purification of this protein from L. otakiensis JCM 15040. Because no specific ligand that has high affinity for this enzyme has been identified, the purification was performed using ...
[摘要] 在异质发酵细菌物种,例如otakiensis的乳酸杆菌(Lactobacillus otakiensis)的培养基中观察到D-亮氨酸,D-异亮氨酸 - 异亮氨酸和D-缬氨酸的显着积累。 otakiensis)JCM 15040.从细菌细胞中纯化导致这种积累的消旋酶,异亮氨酸2-差向异构酶。这是首次报道的在乳酸菌中这种D-支链氨基酸的产生的观察结果,以及在任何生物体中具有异亮氨酸2-差向异构酶活性的消旋酶的第一个实例。在所述的方案中,我们介绍从L中纯化该蛋白质的方法。因为没有鉴定到对该酶具有高亲和力的特异性配体,所以使用硫酸铵级分,四种类型的柱层析和制备型Native-PAGE,不使用亲和柱层析进行纯化。我们希望协议将提供有用的信息用于纯化不能容易地使用亲和柱层析纯化的酶。
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