| Preparation of Purified Gram-positive Bacterial Cell Wall and Detection in Placenta and Fetal Tissues
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Author:
Date:
2016-12-05
[Abstract] Cell wall is a complex biopolymer on the surface of all Gram-positive bacteria. During infection, cell wall is recognized by the innate immune receptor Toll-like receptor 2 causing intense inflammation and tissue damage. In animal models, cell wall traffics from the blood stream to many organs in the body, including brain, heart, placenta and fetus. This protocol describes how to prepare purified cell wall from Streptococcus pneumoniae, detect its distribution in animal tissues, and study the tissue response using the placenta and fetal brain as examples.
[摘要] 细胞壁是所有革兰氏阳性菌表面上的复杂生物聚合物。在感染期间，细胞壁被先天免疫受体Toll样受体2识别，引起强烈的炎症和组织损伤。在动物模型中，从血流到体内许多器官（包括脑，心脏，胎盘和胎儿）的细胞壁运输。该协议描述了如何从肺炎链球菌制备纯化的细胞壁，检测其在动物组织中的分布，并且使用胎盘和胎脑作为实例研究组织反应。
关键词： 细胞壁，肽聚糖，细菌炎症，神经增殖，胎儿神经发生，胎盘运输，Toll样受体2配体，肺炎链球菌
/strong>宿主对感染的反应涉及许多细菌组分的识别，包括细胞壁（CW），一种形成所有革兰氏阳性细菌表面的复合大分子。革兰氏阳性细菌的CW由肽聚糖和磷壁酸的共价网络形成。肺炎链球菌是肺炎，败血症和脑膜炎的主要原因，已经成为研究对包括CW在内的革兰氏阳性细菌感染的先天免疫反应的重要模式生物体。 当肺炎链球菌（肺炎球菌）感染时，CW成分在生长期或抗生素诱导的死亡期间从细菌释放，它在血流中循环并穿过细胞屏障，包括胎盘和血脑屏障。 CW组分具有等于或大于完整细菌的炎性活性（Tuomanen等人，1985a和1985b）。 ...
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| Collagen-induced Arthritis: A Model for Murine Autoimmune Arthritis
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Author:
Date:
2015-10-20
[Abstract] Collagen-induced arthritis (CIA) is a common autoimmune animal model used to study rheumatoid arthritis (RA). The development of CIA involves infiltration of macrophages and neutrophils into the joint, as well as T and B cell responses to type II collagen. In murine CIA, genetically susceptible mice (DBA/1J) are immunized with a type II bovine collagen emulsion in complete Freund’s adjuvant (CFA), and receive a boost of type II bovine collagen in incomplete Freund’s adjuvant (IFA) 21 days after the first injection. These mice typically develop disease 26 to 35 days after the initial injection. C57BL/6J mice are resistant to arthritis induced by type II bovine collagen, but can develop arthritis when immunized with type II chicken collagen in CFA, and receive a boost of type II chicken ...
[摘要] 胶原诱导的关节炎（CIA）是用于研究类风湿性关节炎（RA）的常见的自身免疫动物模型。 CIA的发展涉及巨噬细胞和嗜中性粒细胞渗入关节，以及T和B细胞对II型胶原的反应。在鼠CIA中，用完全弗氏佐剂（CFA）中的II型牛胶原乳剂免疫遗传易感小鼠（DBA/1J），并在第一次给药21天后接受不完全弗氏佐剂（IFA）中II型牛胶原的加强免疫注射。这些小鼠通常在初次注射后26至35天发展疾病。 C57BL/6J小鼠对由II型牛胶原蛋白诱导的关节炎具有抗性，但当在CFA中用II型鸡胶原免疫时可产生关节炎，并且在首次注射后21天在IFA中接受II型鸡胶原蛋白的加强。热死亡的结核分枝杆菌H37RA（MT）在CFA中的浓度对于每个菌株也不同。 DBA/1J小鼠发展具有1mg/ml MT的关节炎，而C57BL/6J小鼠需要和3-4mg/ml MT以形成关节炎。 CIA在C57BL/6J小鼠中缓慢发展，并且与DBA/1J小鼠相比，关节炎病例轻度。该方案描述了用II型牛胶原免疫DBA/1J小鼠和用II型鸡胶原免疫C57BL/6J小鼠。
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| Generation and Maturation of Human Monocyte-derived DCs
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Author:
Date:
2014-08-05
[Abstract] Dendritic cells (DC) are antigen-presenting cells, which play a critical role in the regulation of the adaptive immune response. They act as a bridge between the innate and the adaptive immune systems. An approach to study their function and potentiality is to generate DC-like cells by culturing CD14+ monocyte-enriched peripheral blood mononuclear cells (PBMC). In the presence of GM-CSF and IL-4, these cultures give rise to large numbers of DC-like cells. Generating human-DC from PBMC is a useful tool to study biological functions of human DC.
[摘要] 树突状细胞（DC）是抗原呈递细胞，其在适应性免疫应答的调节中起关键作用。 它们作为先天免疫系统和适应性免疫系统之间的桥梁。 研究它们的功能和潜力的方法是通过培养富集CD14 +单核细胞的外周血单核细胞（PBMC）产生DC样细胞。 在GM-CSF和IL-4存在下，这些培养物产生大量的DC样细胞。 从PBMC产生人类DC是研究人类DC的生物学功能的有用工具。
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