| EAE Induction by Passive Transfer of MOG-specific CD4+ T Cells
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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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| Clonal Culture of Mouse Liver Progenitor Cells
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Author:
Date:
2015-10-20
[Abstract] Liver stem/progenitor cells (LPCs) are defined as bipotential cells differentiating into both hepatocytes and cholangiocytes. For analyzing their differentiation potential, clonal culture has been used for LPCs isolated by a cell sorter. In addition, we can use the culture to assess functions of target genes on differentiation potential of LPCs. This protocol describes the process of cell isolation and colony assay to examine proliferative and differentiation potential of LPCs.
[摘要] 肝干/祖细胞（LPC）被定义为分化成肝细胞和胆管细胞的双能细胞。 为了分析其分化潜力，克隆培养已经用于通过细胞分选仪分离的LPC。 此外，我们可以使用文化来评估目标基因的功能分化潜力的LPCs。 该协议描述细胞分离和集落分析的过程，以检查增殖和分化潜力的LPCs。
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| Hepatitis C virus Cell-to-cell Spread Assay
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Author:
Date:
2014-12-20
[Abstract] Hepatitis C virus (HCV) can infect naïve cells via entry of “cell-free” extracellular virus or direct “cell-to-cell” transmission. Here, we describe an assay for detecting HCV cell-to-cell transmission, using a non-growing cell culture system that avoids confounding effects of cell growth. The assay consists of infecting a small number of cells in a confluent monolayer and then blocking subsequent cell-free extracellular virions with a neutralizing antibody such that only cell-to-cell transmission may occur. Under these conditions, incubation at 37 °C results in the formation of infected cell foci. The extent of cell-to-cell spread can then be determined by counting the number of cells in each focus. The assay may be modified to assess the effects of inhibitors and/or specific cellular ...
[摘要] 丙型肝炎病毒（HCV）可通过"无细胞"细胞外病毒的进入或直接的"细胞到细胞"传递而感染初始细胞。 在这里，我们描述检测HCV细胞到细胞传播，使用一个非增长的细胞培养系统，避免细胞生长的混杂效应的测定。 该测定法包括感染汇合单层中的少量细胞，然后用中和抗体阻断随后的无细胞的细胞外病毒粒子，使得仅可发生细胞与细胞的传递。 在这些条件下，在37℃下孵育导致感染的细胞病灶的形成。 然后可以通过计数每个焦点中的细胞数量来确定细胞与细胞扩散的程度。 可以修改测定以评估抑制剂和/或特异性细胞基因对HCV的细胞与细胞扩散的影响。
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