| Assessing in vitro and in vivo Trogocytosis By Murine CD4+ T cells
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Author:
Date:
2020-05-05
[Abstract] Recognition of antigens by lymphocytes (B, T, and NK) on the surface of an antigen-presenting cell (APC) leads to lymphocyte activation and the formation of an immunological synapse between the lymphocyte and the APC. At the immunological synapse APC membrane and associated membrane proteins can be transferred to the lymphocyte in a process called trogocytosis. The detection of trogocytosed molecules provides insights to the activation state, antigen specificity, and effector functions and differentiation of the lymphocytes. Here we outline our protocol for identifying trogocytosis-positive CD4+ T cells in vitro and in vivo. In vitro, antigen presenting cells are surface biotinylated and pre-loaded with magnetic polystyrene beads before incubating for ...
[摘要] [摘要 ] 抗原呈递细胞（APC）表面的淋巴细胞（B，T和NK）识别抗原会导致淋巴细胞活化并在淋巴细胞和APC之间形成免疫突触。在免疫突触处APC膜和相关的膜蛋白可以转移调用Trogocytosis到淋巴细胞的过程。检测Trogocytosed分子提供见解的激活状态，抗原特异性和效应器功能和差异的淋巴细胞。这里我们列出了我们的协议，用于识别Trogocytosis CD4阳性Tasu 性T细胞在体外和体内。体外，抗原呈递细胞是表面 生物素化并预装磁性聚苯乙烯珠，然后与体外活化的CD4 + T细胞胚细胞（90分钟）或幼稚T细胞（3-24小时）短时间孵育。阳性（trog + ）细胞可通过链霉亲和素染色来筛选经生物素化的经转钙蛋白的APC膜蛋白，然后立即或在随后的孵育后通过流式细胞仪分析其激活表型，效应子功能和效应子的分化，例如，可以鉴定出嗜光细胞的阳性细胞。以前的研究已经描述了分析T细胞嗜光性的方法，以鉴定抗原特异性细胞或被细胞识别的抗原表位。使用当前方案，嗜光性对单个T细胞的影响或能力trog + T细胞调节其他免疫细胞的激活和功能的能力可在一个扩展范围内进行评估 ed时间段。
[背景 ] Trogocytosis是质膜和膜相关分子的细胞间转移。这种现象已在免疫细胞相互作用的分析中得到了广泛研究，并已观察到包括向CD4 +的转移（Wetzel 等，2005； ...
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| Naïve T Lymphocyte Infection by Murine Stem Cell Virus (MSCV) Protocol
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Author:
Date:
2011-11-20
[Abstract] This assay can be used for studying genes related to lymphocyte proliferation and differentiation in vitro. Stimulating lymphocytes to proliferate is important for the infection efficiency of MSCV. After overnight culture, if naïve cells proliferate extensively, the following spin infection will have high efficiency. After infection, cells are ready for the following assay as they will be undergoing proliferation or differentiation.
[摘要] 该测定法可用于体外研究与淋巴细胞增殖和分化相关的基因。 刺激淋巴细胞增殖对于MSCV的感染效率是重要的。 在过夜培养后，如果初始细胞广泛增殖，以下自旋感染将具有高效率。 感染后，细胞准备用于以下测定，因为它们将经历增殖或分化。
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| In vitro Differentiation of Mouse Th0, Th1 and Th2 from Naïve CD4 T Cells
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Author:
Date:
2011-11-20
[Abstract] In vitro differentiation of helper T cells of various lineages is frequently used in T helper cell study. Naïve CD4 T cells can differentiate into certain lineage of T help cells in vitro in the presence of specific stimulatory cytokines and inhibition of cytokines that are essential for the differentiation of other lineages.
[摘要] 在T辅助细胞研究中经常使用各种谱系的辅助T细胞的体外分化。 初始CD4T细胞可以在特异性刺激性细胞因子的存在下分化成T辅助细胞的某些谱系，并抑制对其他谱系分化必不可少的细胞因子。
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