NK cell-based approach for screening novel functional immune genes.
NK cell-based approach for screening novel functional immune genes.
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基于 NK 细胞的方法用于筛选新型功能性免疫基因。
DOI：
10.1016/j.intimp.2010.12.003
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发表时间：
2011-02
期刊：
影响因子：
--
通讯作者：
Zhang Jian
中科院分区：
文献类型：
--
作者：
Wu Longyan;Zhang Cai;Tian Zhigang;Zhang Jian
人类基因组计划为发现新型功能性免疫基因提供了广泛的机会。为了从cDNA文库中找到可能调节NK细胞功能的先天免疫基因，我们使用NK细胞系NK-92作为筛选候选基因的平台。与其他基因转移方法进行比较后，选择电穿孔作为将含有候选基因的cDNA表达质粒导入NK-92细胞的最佳基因转染方法。当转移的基因在NK-92细胞中稳定表达时，检查NK-92细胞的功能变化，包括细胞毒性、溶细胞分子、细胞因子产生和增殖。从100多个候选基因中筛选出2个新基因作为调节NK细胞功能的功能基因，并以NK-92细胞的增殖和细胞毒性为主要指标。随后进行广泛的流式细胞术分析和 RT-PCR。初步数据表明，这两个新基因通过抑制多种激活受体和免疫功能基因的表达，对 NK-92 细胞的细胞毒性产生负面影响。因此，我们描述了一种通过使用 NK 细胞系作为筛选平台来发现 NK 细胞中新功能基因的有效方法。
The human genome project provides extensive opportunities for the discovery of novel functional immune genes. In order to find innate immune genes that might regulate the function of NK cells from a cDNA library, we used an NK cell line, NK-92, as a platform to screen candidate genes. After comparing with other gene transfer methods, electroporation was selected as the best gene transfection approach to deliver cDNA expression plasmids containing candidate genes into the NK-92 cells. When the transferred gene was stably expressed in NK-92 cells, the functional changes in the NK-92 cells were examined, including cytotoxicity, cytolytic molecules, cytokine production, and proliferation. Two novel genes were selected as functional genes that regulate NK cell function from among more than 100 candidate genes, for which the proliferation and cytotoxicity of NK-92 cells were examined as primary indicators. This was followed by extensive flow cytometry analysis and RT-PCR. The primary data indicated that the two novel genes negatively influenced the cytotoxicity of NK-92 cells by inhibiting the expression of several activating receptors and immune functional genes. Therefore, we describe an efficient method for the discovery of novel functional genes in NK cells by using an NK cell line as a screening platform.
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影响因子：
4.2
作者：
Jian Zhang;R. Sun;Haiming Wei;Jianhua Zhang;Z. Tian
通讯作者：
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影响因子：
11.4
作者：
J. Gong;G. Maki;H. Klingemann
通讯作者：
J. Gong;G. Maki;H. Klingemann
影响因子：
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作者：
Linjie Tian;Pingzhang Wang;Jinhai Guo;Xinyu Wang;Weiwei Deng;Chenying Zhang;Dongxu Fu;Xia Gao;T. Shi;D. Ma
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Linjie Tian;Pingzhang Wang;Jinhai Guo;Xinyu Wang;Weiwei Deng;Chenying Zhang;Dongxu Fu;Xia Gao;T. Shi;D. Ma
影响因子：
56.9
作者：
Wu, J;Song, YL;Phillips, JH
通讯作者：
Phillips, JH
影响因子：
5
作者：
Awasthi, A.;Samarakoon, A.;Malarkannan, S.
通讯作者：
Malarkannan, S.