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Development of analytical tools for immune system of Common Marmoset

机译：普通Mar猴免疫系统分析工具的开发

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摘要

Common Marmoset (Callithrix jacchus) is expected to have the immune system similar to human's, but specific tools such as monoclonal antibodies (mAb) have not prepared extensively. In this study, we performed cDNA cloning and sequencing of surface molecules and cytokines expressed mainly on marmoset T cells to prepare transfectants and mAbs, and analyzed cytokine production induced by Th cell stimulation. (1) Marmoset total RNA was extracted from spleen cells stimulated with ConA or TSST-1. The cDNA was amplified using primers with human sequences and used for sequence analysis. (2) Marmoset cDNA in pCX-GFP was transfected to CHO cells and used for mAb preparation. (3) CD4 T cells purified by MACS system from marmoset spleen stimulated via TCR for 24 hrs were submitted for the cytokine analysis in the mRNA level using primers prepared according to the cDNA sequences. As results, 34 cDNA sequences, 6 transfectants and mAbs were obtained. Marmoset spleen T cells enhanced 1L-4, IL-5, TGF- β, TL-10, IFN- γ, TNF- α, IL-2, IL-17A and IL-17F mRNA expression after TCR stimulation, suggesting the similarity of marmoset T cell response to that of humans.
机译：普通Mar猴（Callithrix jacchus）的免疫系统有望与人类相似，但诸如单克隆抗体（mAb）之类的特定工具尚未广泛制备。在这项研究中，我们对主要在mar猴T细胞上表达的表面分子和细胞因子进行了cDNA克隆和测序，以制备转染子和mAb，并分析了Th细胞刺激诱导的细胞因子产生。 （1）从受ConA或TSST-1刺激的脾细胞中提取Mar猴总RNA。使用具有人类序列的引物扩增cDNA，并用于序列分析。 （2）将pCX-GFP中的Mar猴cDNA转染到CHO细胞中，用于制备mAb。 （3）使用MACS系统从通过TCR刺激24小时的24猴脾中纯化的CD4T细胞被提交用于使用根据cDNA序列制备的引物以mRNA水平进行细胞因子分析。结果，获得了34个cDNA序列，6个转染子和mAb。 T猴脾脏T细胞在TCR刺激后增强了1L-4，IL-5，TGF-β，TL-10，IFN-γ，TNF-α，IL-2，IL-17A和IL-17F mRNA的表达，提示它们的相似性。 mo对人的T细胞反应。

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