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Author:
Date:
2016-02-05
[Abstract] In response to exposure to antigen, T cells whose T cell receptor (TCR) are capable of recognizing the self MHC-antigen derived peptide complex, respond to the antigen and differentiate into one of several subsets, namely TH1, TH2, TH17, Treg, etc. characterized by the signature cytokine they secrete, namely IFN-γ, IL-4, IL-17 or IL-10, respectively, referred to as syngeneic polarization as the MHC presenting the foreign antigen/epitope is self-derived.
T cell responses following incubation for defined periods, usually 3 days for mouse splenocytes, are routinely measured by assessing the antigen-stimulated proliferation of T cells by measuring the radiolabeled precursor thymidine incorporated into the genomic DNA of the dividing T cell; the direction of polarization ...
[摘要] 响应于暴露于抗原，其T细胞受体（TCR）能够识别自身MHC抗原衍生的肽复合物的T细胞应答抗原，并分化为几个亚类之一，即TH1，TH2，TH17，Treg， 等。其特征在于它们分泌的特征性细胞因子，即IFN-γ，IL-4，IL-17或IL-10，分别称为同基因极化，因为呈递外源抗原/表位的MHC是自身衍生的。通常通过测量掺入分裂T细胞的基因组DNA中的放射性标记的前体胸腺嘧啶，通过评估抗原刺激的T细胞增殖来常规测量对于确定的时间段（通常为小鼠脾细胞3天）孵育后的T细胞应答;通过使用培养上清液的ELISA或通过细胞内细胞因子染色随后通过流式细胞术测量由增殖或非增殖应答性T细胞产生的细胞因子来评估极化方向。在下面详述的方案中，我们描述使用的同源小鼠骨髓源性树突状细胞（BMDC）作为APC刺激脾源性T细胞。通过将放射性标记的前体胸苷掺入基因组DNA中来测量T细胞的增殖反应，并通过在72小时期间测量它们分泌的细胞因子即IFN-γ，IL-4和IL-17来评估其极化方向使用ELISA。此外，我们在细胞内细胞因子染色后使用流式细胞术检测CD3 + /CD4 + /CD25低人群中的IL-17阳性T细胞。使用牛分枝杆菌（Mycobacterium bovis） - Bacille Calmette ...
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Author:
Date:
2014-11-20
[Abstract] Tumors are heterogeneous microenvironments where complex interactions take place between neoplastic cells and infiltrating inflammatory cells, such as tumor-associated macrophages (TAM) and tumor-associated dendritic cells (TADC). The relevance of tumor-infiltrating mononuclear myeloid cells is underscored by clinical studies showing a correlation between their abundance and poor prognosis (Laoui et al., 2011). These cells are able to promote tumor progression via several mechanisms, including induction of angiogenesis, remodeling of the extracellular matrix, stimulation of cancer cell proliferation and metastasis and the inhibition of adaptive immunity (Laoui et al., 2011). Moreover, mononuclear myeloid cells are characterized by plasticity and versatility in response ...
[摘要] 肿瘤是异质微环境，其中在肿瘤细胞和浸润性炎症细胞例如肿瘤相关巨噬细胞（TAM）和肿瘤相关树突细胞（TADC）之间发生复杂的相互作用。临床研究强调了肿瘤浸润性单核骨髓细胞的相关性，显示其丰度与不良预后之间的相关性（Laou等人，2011）。这些细胞能够通过几种机制促进肿瘤进展，包括诱导血管生成，重塑细胞外基质，刺激癌细胞增殖和转移以及抑制适应性免疫（Laoui等人，2011 ）。此外，单核骨髓细胞的特征在于响应于微环境信号的可塑性和通用性，导致不同的活化状态，如通过在肿瘤中存在不同的功能性TAM亚型所说明的（Movahedi等人，2010; Laoui et al。，2014）。在这里，我们描述了允许其分子和功能表征的TAM和TADC的有价值的分离技术。
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