| In vivo DCs Depletion with Diphtheria Toxin and MARCO+/MOMA1+ Cells Depletion with Clodronate Liposomes in B6.CD11c-DTR Mice
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Author:
Date:
2016-08-05
[Abstract] To evaluate precisely the relative roles of different splenic phagocytic cells during an immune response, efficient methods for the depletion of specific populations are needed. Here, we describe the protocols for the depletion of splenic dendritic cells (DCs) by human diphtheria toxin (DTx) treatment in target mice (which express the human DTx receptor in all CD11c+ DCs) and for the specific depletion of MARCO+/MOMA-1+ marginal zone macrophages (MZMΦs) with clodronate liposomes (ClLip) treatment (when a small dose of ClLip is ministered, MZMΦs preferentially uptake ClLip, and clodronate is released inside those cells causing apoptosis-mediated cell death). These protocols are adaptations from previous works (Jung et al., 2002; McGaha et al ...
[摘要] 为了准确地评估不同的脾吞噬细胞在免疫应答期间的相对作用，需要用于消耗特定群体的有效方法。 在这里，我们描述了通过人类白喉毒素（DTx）治疗在目标小鼠（其在所有CD11c + DC中表达人类DTx受体）中脾脏树突状细胞（DCs） 用氯膦酸脂质体（ClLip）处理（当小剂量的ClLip被分配时，MZMΦs优先摄取）的MARCO + /MOMA-1 + 边缘区巨噬细胞（MZMΦ） ClLip和氯膦酸盐在那些细胞内释放，引起凋亡介导的细胞死亡）。 这些方案是来自先前作品的改编（Jung等人，2002; McGaha等人，2011），并且用于评价DC和MZMΦ的各自的作用 在实验性血液阶段疟疾感染的急性期期间（Borges da Silva等人，2015）。
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| Isolation of Splenic Dendritic Cells Using Fluorescence-activated Cell Sorting
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Author:
Date:
2015-03-05
[Abstract] The spleen is a vastly vasculated organ and consists of a complex organized network of innate and adaptive immune cells. This permits the specialized functions of the spleen such as antibacterial and antifungal immunity and iron metabolism among others (Mebius and Kraal, 2005). Different dendritic cell (DC) subsets reside in the spleen and can be defined by the expression of unique surface markers. These DC subsets are recognized to perform non-redundant functions in the immune system (Merad et al., 2013). In our recent study, we found that Inositol Requiring Enzyme (IRE)-1 is specifically activated in splenic CD8a+ DCs. Furthermore, loss of X-box binding protein (XBP)-1 – the transcription factor regulated by IRE-1 – resulted in defective cross-presentation of dead ...
[摘要] 脾脏是一个巨大的血管的器官，由一个复杂的有组织的网络的先天和适应性免疫细胞组成。 这允许脾的特殊功能，例如抗菌和抗真菌免疫和铁代谢等（Mebius和Kraal，2005）。 不同的树突状细胞（DC）子集驻留在脾脏中，并且可以通过表达独特的表面标志物来定义。 认识到这些DC亚类在免疫系统中执行非冗余功能（Merad等人，2013）。 在我们最近的研究中，我们发现肌醇需求酶（IRE）-1在脾CD8a + DC中特异性激活。 此外，X-box结合蛋白（XBP）-1（由IRE-1调节的转录因子）的缺失导致脾CD8a + DC（Osorio em> et al。，2014）。 该协议允许分离用于实验使用的特异性DC亚群ex-vivo 。
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