| In vivo Analysis of Neutrophil Infiltration during LPS-induced Peritonitis
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Author:
Date:
2016-10-05
[Abstract] Bacterial lipopolysaccharide (LPS) is present in the outer membrane of Gram-negative bacteria and functions as pathogen-associated molecular pattern (PAMP) (Whitfield and Trent, 2014). LPS therefore is a potent activator of inflammatory responses leading to cytokine release and neutrophils recruitment. The lipid A moiety of LPS activates the complex consisting of the LPS binding protein (LBP), CD14, MD-2 and Toll-like receptor 4 (TLR4) and the non-canonical inflammasome-linked caspases-4, 5 and 11, which in turn activate the canonical NLRP3 inflammasome (Shi et al., 2014; Hagar et al., 2013; Kayagaki et al., 2013; Hoshino et al., 1999; Poltorak, 1998; Nagai et al., 2002; Park et al., 2009; Ratsimandresy et al., 2013). In ...
[摘要] 细菌脂多糖（LPS）存在于革兰氏阴性细菌的外膜中，并且作为病原体相关分子模式（PAMP）（Whitfield和Trent，2014）。因此LPS是炎症反应的有效活化剂，导致细胞因子释放和嗜中性粒细胞募集。 LPS的脂质A部分激活由LPS结合蛋白（LBP），CD14，MD-2和Toll样受体4（TLR4）组成的复合物和非规范的炎症小体连接的半胱天冬酶-4,5和11反过来激活典型的NLRP3炎症小体（Shi等人，2014; Hagar等人，2013; Kayagaki等人，2013; Hoshino等人，1999; Poltorak，1998; Nagai等人，2002; Park等人，2009; Ratsimandresy等人， et al 。，2013）。特别地，响应于炎症小体激活产生的细胞因子白细胞介素（IL）-1β在通过促进嗜中性粒细胞粘附和迁移的中性粒细胞募集中具有关键作用（McDonald等人，2010）。该方案允许研究的由LPS诱导的炎症反应，其通过跟踪体内髓过氧化物酶（MPO）活性来影响嗜中性粒细胞浸润（de Almeida等人，2015）。
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| Differentiation of THP1 Cells into Macrophages for Transwell Co-culture Assay with Melanoma Cells
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Author:
Date:
2015-11-05
[Abstract] Understanding how immune cells such as macrophages interact with cancer cells is of increasing interest, as cancer treatments move towards combing both targeted- and immuno-therapies in new treatment regimes. This protocol is using THP-1 cells, a human leukemia monocytic cell line that can be differentiated into macrophages. This allows studying the effects of the macrophage secretome on cancer cells (on e.g., growth, drug response or gene expression) in co-cultures without direct cell contact interactions. This is an important aspect as it removes the presence of any phagocytic aspect to changes in the cancer cell number and behaviour. The in vitro THP-1 monocyte differentiation into polarized macrophages was used to study the effects of both M1 and M2 type populations ...
[摘要] 了解免疫细胞如巨噬细胞如何与癌细胞相互作用越来越感兴趣，因为癌症治疗趋向于在新的治疗方案中结合靶向和免疫治疗。该协议使用THP-1细胞，人类白血病单核细胞系，可以分化成巨噬细胞。这允许在没有直接细胞接触相互作用的共培养物中研究巨噬细胞分泌蛋白组织对癌细胞的作用（例如生长，药物反应或基因表达）。这是一个重要的方面，因为它消除了任何吞噬方面的癌细胞数量和行为的变化的存在。将体外THP-1单核细胞分化为极化巨噬细胞用于研究巨噬细胞的M1和M2型群体对黑素瘤细胞的作用（Smith等人，2014年， ; Tsuchiya等人，1980）。 M1型巨噬细胞通常被认为是与M2型巨噬细胞相反的肿瘤抑制，M2型巨噬细胞被认为具有组织修复和促进肿瘤生长的活性。
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| Macrophage Infection by Dimorphic Fungi
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Author:
Date:
2012-07-20
[Abstract] Mammalian infection by dimorphic fungi occurs through the inhalation of asexual spores (conidia), which are phagocytosed by host pulmonary alveolar macrophages of the innate immune system. Once phagocytosed, fungal conidia germinate into the pathogenic cell type; unicellular yeast cells which divide by fission (Vanittanakom et al., 2006; Boyce et al., 2011). To investigate if mutation of a particular fungal gene affects macrophage phagocytosis or the production of yeast cells, a murine macrophage cell culture assay can be utilized. This protocol was developed for Penicillium marneffei but is applicable to most dimorphic fungi.
[摘要] 两形态真菌的哺乳动物感染通过吸入无性孢子（分生孢子）发生，所述无性孢子被先天免疫系统的宿主肺泡巨噬细胞吞噬。 一旦被吞噬，真菌分生孢子萌发成致病细胞类型; 通过裂变分裂的单细胞酵母细胞（Vanittanakom et al。，2006; Boyce et al。，2011）。 为了研究特定真菌基因的突变是否影响巨噬细胞吞噬作用或酵母细胞的产生，可以利用鼠巨噬细胞培养测定法。 该方案是为马尼埃青霉开发的，但适用于大多数二形真菌。
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