%0 Journal Article
%T Δ42PD1稳定表达细胞系的建立及其功能的初步研究<br>Generation of Δ42PD1-Expressing Stable Cell line and Preliminary Research on the Function Thereof
%A 程林
%A 唐娴
%A 曹廷智
%A 徐六妹
%A 彭巧丽
%A 何云
%A 王辉
%J Immunology Studies
%P 1-7
%@ 2330-1678
%D 2016
%I Hans Publishing
%R 10.12677/IS.2016.41001
%X <div style=\"text-align:justify;\">
	<span style=\"line-height:1.5;\">目的：建立稳定表达Δ42PD1的293T细胞系，并探索其对AKT，NF-κB和Erk1/2磷酸化的影响。方法：分别将含有人PD1和Δ42PD1基因的真核表达质粒稳定转染293T细胞，用流式细胞术筛选稳定细胞系，用RT-PCR和Western blot进一步鉴定目基因的表达。将稳定细胞系分别于PBMCs进行孵育，以激活Δ42PD1和PD1，用流式细胞术检测细胞内AKT，NF-κB和Erk1/2的磷酸化水平。结果：通过流式细胞术筛选出稳定表达PD1和Δ42PD1的293T细胞系，RT-PCR和Western blot确认了目的基因的表达。与PBMC混合孵育后，293T细胞内AKT的磷酸化水平被Δ42PD1或PD1显著抑制，而NF-κB和Erk1/2的磷酸化水平没有显著变化。结论：本研究发现Δ42PD1与PD1的功能类似，可能是一种重要的抑制性免疫调节受体。 <br />
Objective: To establish Δ42PD1-expressing stable 293T cell line and explore the effect of Δ42PD1 on phosphorylation of AKT, NF-κB and Erk1/2. Methods: The human PD1- and Δ42PD1-carrying eukaryotic expression plasmids were stably transfected into 293T cells respectively. Stable cell lines were screened using flow cytometry, and confirmed by RT-PCR and Western blot. The cell lines were co-cultured with PBMC to activate PD1 and Δ42PD1, and then analyzed the phosphorylation of AKT, NF-κB and Erk1/2 via flow cytometry. Results: We obtained PD1- and Δ42PD1-stably expressing 293T cell lines by flow cytometry. The expression of genes of interest was confirmed by RT-PCR and Western blot. After co-culture with PBMC, phosphorylation of AKT but not NF-κB or Erk1/2 in 293T cells was significantly inhibited by Δ42PD1 or PD1. Conclusion: The present study found that functionally similar to PD1, Δ42PD1 could be an important inhibitory immune-regula- tory receptor.</span><span style=\"line-height:1.5;\"></span> 
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%K 程序性细胞死亡受体1，Delta-42程序性细胞死亡受体1，流式细胞术，蛋白激酶B <br>PD1
%K Δ42PD1
%K Flow Cytometry
%K AKT
%U http://www.hanspub.org/journal/PaperInformation.aspx?PaperID=18390