| Murine Liver Myeloid Cell Isolation Protocol
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Author:
Date:
2015-05-20
[Abstract] In homeostasis, the liver is critical for the metabolism of nutrients including sugars, lipids, proteins and iron, for the clearance of toxins, and to induce immune tolerance to gut-derived antigens. These functions predispose the liver to infection by blood-borne pathogens, and to a variety of diseases ranging from toxin and medication-induced disorders (CCl4, acetaminophen) to metabolic disorders (steatohepatitis, alcoholic liver disease, biliary obstruction, cholestasis) or autoimmunity. Chronic liver injury often progresses to life threatening fibrosis and can end in liver cirrhosis and hepatocellular carcinoma (Pellicoro et al., 2014).
The liver contains parenchymal cells or hepatocytes that make up the majority of hepatic cells. It also contains ...
[摘要] 在内环境稳定中，肝脏对于营养物质（包括糖，脂质，蛋白质和铁）的代谢是关键的，用于清除毒素，并诱导对肠衍生的抗原的免疫耐受。这些功能使肝脏感染由血源性病原体引起的感染，并且导致各种疾病，从毒素和药物诱导的疾病（CCl 4，对乙酰氨基酚）到代谢性疾病（脂肪性肝炎，酒精性肝病，胆汁阻塞，胆汁淤积）或自身免疫。慢性肝损伤常常进展为威胁生命的纤维化，并且可以在肝硬化和肝细胞癌中结束（Pellicoro等人，2014）。
肝脏含有构成肝细胞大部分的实质细胞或肝细胞。它还含有非实质结构细胞，例如窦状内皮细胞和大量非实质的先天免疫细胞，主要是单核细胞，嗜中性粒细胞，巨噬细胞，DC，NK和NKT细胞，其在感染的情况下可触发适应性免疫应答或其他致病性侮辱（Jenne和Kubes，2013）。如何调节这种免疫应答决定了急性和慢性肝损伤的程度（Stijlemans等人，2014）。在这种情况下，肝巨噬细胞已经被证明在肝损伤中发挥中心但发散（从启动到分解）功能。（Sica等人，2014）。在过去几年中已经变得清楚的是，肝巨噬细胞由两类，组织驻留巨噬细胞，源自卵黄囊/胎儿肝祖细胞的库普弗细胞（KC）和来源于骨髓衍生的Ly6C的组织浸润巨噬细胞Hi 单核细胞（Jinhoux和Jung，2014; ...
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| Isolation of Particles of Recombinant ASC and NLRP3
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Author:
Date:
2015-05-20
[Abstract] NLRP3 inflammasome is a multiprotein complex responsible for the activation of inflammatory caspase-1, resulting in processing and release of pro-inflammatory cytoquines IL-1β and IL-18 (Schroder and Tschopp, 2010). This inflammasome is composed of the sensor protein NLRP3 connected to caspase-1 through the adaptor protein ASC (apoptosis-associated speck-like protein with a caspase-recruitment domain) (Schroder and Tschopp, 2010). We and others have reported that upon inflammasome activation functional oligomeric inflammasome particles of NLRP3 and ASC were released from cells, acting as danger signals to amplify inflammation by promoting the activation of caspase-1 extracellularly (Baroja-Mazo et al., 2014; Franklin et al., 2014).
Studying the extracellular ...
[摘要] NLRP3炎症小体是负责炎症半胱天冬酶-1活化的多蛋白复合物，导致促炎细胞因子IL-1β和IL-18的加工和释放（Schroder和Tschopp，2010）。这种炎症小体由通过衔接蛋白ASC（凋亡相关的斑点样蛋白与半胱氨酸蛋白酶募集结构域）连接到caspase-1的传感蛋白NLRP3组成（Schroder和Tschopp，2010）。我们和其他人已经报道，在炎症小体激活时，NLRP3和ASC的功能性寡聚炎症小体颗粒从细胞中释放，充当危险信号，通过促进细胞外caspase-1的活化来扩增炎症（Baroja-Mazo等，
通过纯化ASC的重组颗粒或组成型激活的NLRP3突变体来研究寡聚ASC和NLRP3炎症小体颗粒的细胞外功能是可能的与相关的cryopyrin相关周期性综合征（CAPS，突变p.D303N），都标记有黄色荧光蛋白（YFP），并在HEK293细胞中表达。通过重组ASC或突变体NLRP3在HEK293细胞中的表达导致它们自发聚集成斑点（Baroja-Mazo等人，2014）的事实促进了纯化过程，并且方案最初从Fernandes-Alnemri和Alnemri（2008年）。
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| Isolation and in vitro Activation of Mouse Peyer’s Patch Cells from Small Intestine Tissue
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Author:
Date:
2014-11-05
[Abstract] The lumen of gastrointestinal tract is exposed to several potentially pathogenic microorganisms, thus it is extremely relevant to understand how immunosurveilance can be established. Peyer’s Patches (PPs) are oval or round lymphoid nodules that protrude from the outer wall of the ileum portion of small intestine. PPs contain a high percentage of B and T lymphocytes, macrophages and dendritic cells. Here we summarize a protocol for isolation and culture of mouse PP cells, which can be used to get a better insight into immunopathologies of microbes and to evaluate immune responses elicited by mucosal vaccines.
[摘要] 胃肠道的腔暴露于几种潜在的致病微生物，因此，了解如何建立免疫耐受是非常重要的。 派尔氏贴剂（PP）是椭圆形或圆形淋巴结，其从小肠的回肠部分的外壁突出。 PP含有高百分比的B和T淋巴细胞，巨噬细胞和树突细胞。 在这里我们总结一个协议的小鼠PP细胞的分离和培养，可用于更好地了解微生物的免疫病理学和评估由粘膜疫苗引发的免疫反应。
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