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加湿CO2培养箱

Company: Thermo Fisher Scientific
Catalog#: BB15
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Differentiation of Myeloid-derived Suppressor Cells from Murine Bone Marrow and Their Co-culture with Splenic Dendritic Cells
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Date:
2017-09-20
[Abstract]  Myeloid-derived suppressor cells (MDSCs) possess the ability to suppress the immune response, and to amplify the regulatory properties of other immune cells, i.e., dendritic cells. Here we describe a protocol in which MDSCs were differentiated from murine bone marrow cells, and CD11c+ dendritic cells were purified from murine spleens. MDSCs and CD11c dendritic cells can be co-cultured and the immunoregulatory phenotype of the MDSCs-conditioned dendritic cells could be assessed by means of a specific functional in vivo experiment, i.e., a skin test as a measure of the delayed-type hypersensitivity reaction toward a poorly immunogenic antigen. [摘要]  骨髓来源的抑制细胞(MDSCs)具有抑制免疫应答的能力,并扩增其他免疫细胞即树突状细胞的调节特性。 在这里,我们描述了MDSC与鼠骨髓细胞分化的方案,并且从鼠脾中纯化CD11c +树突状细胞。 可以共培养MDSC和CD11c树突状细胞,并且可以通过特定的功能体内实验来评估MDSCs条件树突细胞的免疫调节表型,即皮肤试验作为延迟型超敏反应的量度 抗免疫原性较差的抗原。
【背景】骨髓来源的抑制细胞(MDSCs)是由早期分化阶段的巨噬细胞,粒细胞,树突状细胞和骨髓细胞的前体组成的骨髓细胞组(Youn等人,2008),其在肿瘤的淋巴组织中大量积累感染性小鼠以及感染性疾病,败血症和创伤的小鼠。这些细胞的主要特征是它们以Ag特异性和/或非特异性方式抑制T细胞应答的能力。这些细胞现在被认为是负责肿瘤相关免疫缺陷的主要细胞类型之一;涉及MDSC介导的免疫抑制的主要因素包括Arg1的高表达(Marvel和Gabrilovich,2015)。精氨酸酶1(Arg1)和吲哚胺2,3-双加氧酶1(IDO1)分别是催化L-精氨酸(L-Arg)和L-色氨酸(L-Trp)降解的免疫调节酶,导致局部氨基酸剥夺。此外,与Arg1不同,IDO1在树突细胞(DC)中也具有非酶信号传导活性(Mondanelli等,2017)。除了其固有的免疫抑制活性外,MDSC还可能扩增其他免疫细胞的调节特性,特别是在肿瘤微环境中。虽然建立了MDSC-巨噬细胞相互作用的一些机制(Ugel等,2015),MDSCs和DCs之间的串扰仍然不清楚(Ostrand-Rosenberg等,2012);为弥补这一差距,我们已经制定了该方案,并且我们证明了Arg1 ...

Construction of FWPV Chimaeric MVA
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Date:
2015-01-05
[Abstract]  Construction of chimaeric MVA is a useful tool with which to study gene function of related viruses. The protocol given here describes MVA chimaeras containing genes from Fowlpox virus (FWPV), although this can be applied to DNA derived from other organisms. There are a number of steps required to make the chimaeric MVA: 1) Purification of viral particles; 2) Extraction of DNA from purified viral particles; 3) Assembly of linear recombination templates; 4) Transfection of linear recombination templates; 5) Selection of chimaeric MVA.
Note: This procedure uses live virus, and should be conducted using Good Microbiological Practice, in accordance with international and national biocontainment requirements. This procedure also involves Genetic Modification of microorganisms, and ...
[摘要]  嵌合MVA的构建是研究相关病毒的基因功能的有用工具。 本文给出的方案描述了含有来自鸡痘病毒(FWPV)的基因的MVA嵌合体,尽管这可以应用于来自其他生物体的DNA。 制备嵌合MVA需要许多步骤:1)病毒颗粒的纯化; 2)从纯化的病毒颗粒中提取DNA; 3)装配线性重组模板; 4)线性重组模板的转染; 5)嵌合MVA的选择。
注意:此程序使用活病毒,并应根据国际和国家生物承诺要求使用良好微生物实践进行。 该程序还涉及微生物的遗传修饰,并且在开始前应获得适当的安全批准。

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