水生生物学报2018，Vol.42Issue(3)：480-487,8.DOI:10.7541/2018.060
半滑舌鳎外周血T淋巴细胞的分离、鉴定及TCRβ基因免疫应答分析
ISOLATION AND IDENTIFICATION OF T-LYMPHOCYTES FROM PERIPHERAL BLOOD AND IMMUNE RESPONSE ANALYSIS OF TCRβ CHAIN GENE IN CYNOGLOSSUS SEMILAEVIS
摘要
Abstract
The density gradient method was used to isolate lymphocytes from peripheral blood of Cynoglossus semi-laevis,and the isolated lymphocytes were cultured in complete DMEM medium(with 0.3 μg/mL PHA)for 3—4 days. Peripheral blood lymphocytes were separated to non-adherent lymphocytes and adherent lymphocytes using self-made nylon hair column. Non-adherent lymphocytes specifically bind with mouse anti-human FTIC-CD3 monoclonal anti-body,while adherent lymphocytes specifically bind to anti-human FTIC-CD19 monoclonal antibody.TCRβ specifi-cally expressed on non-adherent lymphocytes. Taken together, non-adherent lymphocytes were confirmed as T lympho-cytes.The results of qRT-PCR showed that TCRβ was expressed in all detected 13 tissues with high expression level in the intestine,stomach,spleen,head kidney and the lowest level in muscle.The expression of TCRβ gene was signifi-cantly induced in the liver, spleen and gill with peak level at 72—96h after V. Anguillarum infection. These re-sults indicate that TCRβ gene plays an important role in adaptive cellular immune response.关键词
半滑舌鳎/T淋巴细胞/B淋巴细胞/外周血/TCRβ基因/免疫应答Key words
Cynoglossus semilaevis/T Lymphocytes/B Lymphocytes/Peripheral blood/TCRβ gene/Immune response分类
农业科技引用本文复制引用
汪林庆,王航,陈亚东,杨光,白莉,孙璐明,沙珍霞..半滑舌鳎外周血T淋巴细胞的分离、鉴定及TCRβ基因免疫应答分析[J].水生生物学报,2018,42(3):480-487,8.基金项目
国家自然科学基金(31572644) （31572644）
青岛海洋科学与技术国家实验室海洋渔业科学与食物产出过程功能实验室开放基金(2016LMFS-A25)资助[Supported by the National Natural Science Foundation of China(31572644) （2016LMFS-A25）
Laboratory for Marine Fisheries Science and Food Production Processes,Qingdao National Laboratory for Marine Science and Technology(2016LMFS-A25)] （2016LMFS-A25）
