{{'Search' | translate}}
 

TruStain FcXTM (anti-mouse CD16/32) Antibody

Company: BioLegend
Catalog#: 101319
Bio-protocol()
Company-protocol()
Other protocol()

Evaluation of B Cell Proliferation in vivo by EdU Incorporation Assay
Author:
Date:
2020-05-05
[Abstract]  Generation of antibodies is crucial for establishing enduring protection from invading pathogens, as well as for maintaining homeostasis with commensal bacteria at mucosal surfaces. Chronic exposure to microbiota- and dietary- derived antigens results in continuous production of antibody producing cells within the Peyer’s patch germinal center structures. Recently, we have shown that B cells responding to gut-derived antigens colonize the subepithelial dome (SED) in Peyer’s patches and rapidly proliferate independently of their relative BCR affinity. To evaluate B cell proliferation within different niches in Peyer’s patches, we applied in vivo EdU incorporation assay as described in this protocol.
[摘要]  [摘要] 抗体的产生对于建立持久保护免受入侵的病原体以及在粘膜表面维持共生细菌的稳态至关重要。长期暴露于微生物和饮食来源的抗原可导致Peyer's淋巴瘤中不断产生抗体的细胞最近,我们发现对肠道源性抗原有反应的B细胞在Peyer's斑块中上皮下穹顶(SED)定居并迅速增殖,而与它们的相对BCR亲和力无关。评估Peyer's斑块中不同壁ches中的B细胞增殖。 ,我们按照此方案中所述进行了体内EdU 掺入分析。

[背景] 长寿命抗体生成细胞,也被称为浆或血浆细胞(PC机)主要都来源于生发中心(GCS),显微解剖的网站,表单中的免疫器官感染后或免疫接种。进入GC反应涉及亲和基础的B细胞的竞争,每个细胞都表达对抗原具有特定亲和力和特异性的抗原特异性B细胞受体(BCR),一旦遇到抗原,B细胞就会发生活化,并伴随着通过快速增殖的广泛克隆扩增。斑块(PPs)是位于小肠的淋巴器官,是B细胞类将其免疫球蛋白转换为IgA 的主要部位。上皮下穹S(SED)是PP中的一个小生境,其中免疫细胞(包括B细胞)与在PPs中发生GC前事件期间,带有高亲和力BCR的B细胞在SED的定殖和形成过程中没有表现出优先优势 的PC,表明在该位点没有发生基于亲和力的竞争(Biram et al。,2019),尽管如此,只有高亲和力的克隆进入GC结构并进入生发中心反应。外周淋巴结,脾脏和PP中的GC ...

Characterization of Immunological Niches within Peyer’s Patches by ex vivo Photoactivation and Flow Cytometry Analysis
Author:
Date:
2020-03-20
[Abstract]  T follicular helper (Tfh) cells regulate B cell selection for entry into the germinal center (GC) reaction or for differentiation into antibody forming cells. This process takes place at the border between the T and B zones in lymphoid organs and involves physical contacts between T and B cells. During these interactions, T cells endow the B cells with selection signals that promote GC seeding or plasmablast differentiation based on their B cell receptor affinity. In Peyer’s patches (PPs), T cells promote B cell colonization of the subepithelial dome (SED) without effective affinity-based clonal selection. To specifically characterize the T cell population that resides within the SED niche, we performed ex vivo photoactivation of the SED compartment followed by flow cytometry ... [摘要]  [摘要] T卵泡辅助细胞(Tfh )调节B细胞的选择,使其进入生发中心(GC)反应或分化为抗体形成细胞。此过程发生在淋巴器官的T和B区之间的边界,涉及T和B细胞之间的物理接触。在这些相互作用中,T细胞赋予B细胞选择信号,这些信号根据其B细胞受体亲和力促进GC接种或成浆细胞分化。在Peyer氏斑(PPs)中,T细胞在没有有效亲和力的情况下促进上皮下穹sub (SED)的B细胞定殖。 ed克隆选择。为了具体表征驻留在SED利基空间中的T细胞群体,我们按照本方案中所述对SED隔室进行了离体光激活,然后进行了标记细胞的流式细胞仪分析。该技术将空间和细胞信息整合到了免疫小生境研究中,可以适应各种实验系统。

【背景技术】亲和力成熟是血清抗体对特定抗原的亲和力随时间而增加的过程,这是通过在生发中心(GC)中选择带有高亲和力BCR的B细胞来实现的。增加抗体亲和力通过体细胞超突变和基于亲和力的选择,这由T滤泡辅助(编排的处理的反复循环介导的TFH )细胞奥普雷亚;(开普勒和Perelson,1993 和Perelson,1997; Victora和Nussenzweig, 2012)。GC由两个微观解剖部位组成;暗区(B细胞增殖并获得体细胞超突变)和亮区(B细胞与相关抗原和T细胞相互作用)。尽管活体成像技术能够确定GC中的免疫细胞动力学(Allen 等,2007; ...

Comments