斑点叉尾鮰不同克隆性同种异体抗原依赖性细胞毒性细胞系的建立与分析
Development and analysis of various clonal alloantigen-dependent cytotoxic cell lines from channel catfish.
作者信息
Stuge T B, Wilson M R, Zhou H, Barker K S, Bengtén E, Chinchar G, Miller N W, Clem L W
机构信息
Department of Microbiology, University of Mississippi Medical Center, Jackson, MS 39216, USA.
出版信息
J Immunol. 2000 Mar 15;164(6):2971-7. doi: 10.4049/jimmunol.164.6.2971.
To determine the phenotypes of cytotoxic cells in channel catfish, clonal alloantigen-dependent leukocyte lines were established from mixed leukocyte cultures. Each clone was analyzed for expression of TCR alpha and beta genes by RT-PCR and for target cell specificity by 51Cr-release assay. Based on the above criteria, the following five different cell types were identified among the 19 clones analyzed: 1) TCR alphabeta+ allospecific cytotoxic cells, 2) TCR alphabeta+ nonspecific cytotoxic cells, 3) allospecific TCR alphabeta+ noncytotoxic cells, 4) TCR alphabeta- nonspecific cytotoxic cells, and 5) TCR alphabeta- allospecific cytotoxic cells. The demonstration of cloned, TCR alphabeta+, allospecific cytotoxic effectors provides the strongest evidence to date for the existence of cytotoxic T cells in fish.
为了确定斑点叉尾鮰中细胞毒性细胞的表型，从混合白细胞培养物中建立了克隆的同种异体抗原依赖性白细胞系。通过逆转录聚合酶链反应（RT-PCR）分析每个克隆中TCRα和β基因的表达，并通过51Cr释放试验分析其对靶细胞的特异性。基于上述标准，在分析的19个克隆中鉴定出以下五种不同的细胞类型：1）TCRαβ+同种特异性细胞毒性细胞，2）TCRαβ+非特异性细胞毒性细胞，3）同种特异性TCRαβ+非细胞毒性细胞，4）TCRαβ-非特异性细胞毒性细胞，以及5）TCRαβ-同种特异性细胞毒性细胞。克隆的、TCRαβ+、同种特异性细胞毒性效应细胞的证明为鱼类中细胞毒性T细胞的存在提供了迄今为止最有力的证据。
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