Expression profiles of TCRβ and CD8α mRNA correlate with virus-specific cell-mediated cytotoxic activity in ginbuna crucian carp
Expression profiles of TCRβ and CD8α mRNA correlate with virus-specific cell-mediated cytotoxic activity in ginbuna crucian carp
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DOI：
10.1016/j.virol.2006.01.019
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发表时间：
2006-05
期刊：
影响因子：
3.7
通讯作者：
T. Somamoto;Y. Yoshiura;Atsushi Sato;M. Nakao;T. Nakanishi;N. Okamoto;M. Ototake
中科院分区：
文献类型：
--
作者：
T. Somamoto;Y. Yoshiura;Atsushi Sato;M. Nakao;T. Nakanishi;N. Okamoto;M. Ototake
我们前期的研究表明，银鲫克隆及其同源细胞系可诱导致敏白细胞产生病毒特异性细胞介导的细胞毒作用。在本研究中，我们试图确定鱼的病毒特异性细胞毒性细胞群是否表达CD 8 α和TCRβ基因。银鲫白细胞经免疫磁性分离和密度梯度离心分离后，分为四个组分：组分A，密度为1.08g/ml（主要是淋巴细胞）;组分B，密度为1.08 μ g/ml的sIg阴性白细胞;组分C，sIg阳性细胞级分D，密度为1.08-1.09 μ g/ml的白细胞（主要是嗜中性粒细胞）。来自未感染鱼的所有组分中的白细胞对病毒感染的同系细胞不表现出细胞毒性活性，并且弱表达CD 8 α和TCRβ mRNA。相反，来自病毒感染鱼的组分A和B中的白细胞表现出高水平的细胞毒性活性，并强烈表达CD 8 α和TCRβ mRNA。此外，与病毒感染的靶细胞共培养可上调效应细胞中CD 8 α和TCRβ的mRNA表达，而与未感染的靶细胞共培养则无此作用。目前的研究表明，鱼类具有病毒特异性细胞毒性细胞的表型和基因表达模式相似的哺乳动物中的CTL。
Our previous studies have demonstrated that virus-specific cell-mediated cytotoxicity of sensitized leukocytes can be induced using clonal ginbuna crucian carp and their syngeneic cell lines. In the present study, we attempt to determine if virus-specific cytotoxic cell populations of fish express CD8α and TCRβ genes. Leukocytes from ginbuna crucian carp were separated into four fractions by immunomagnetic separation and density gradient centrifugation: Fraction A, leukocytes with a density of 1.08 g/ml (primarily lymphocytes); Fraction B, sIg-negative leukocytes with density of 1.08 g/ml; Fraction C, sIg-positive cells (primarily B-lymphocytes); Fraction D, leukocytes with a density of 1.08–1.09 g/ml (primarily neutrophils). Leukocytes in all fractions from uninfected fish do not exhibit cytotoxic activity against virus-infected syngeneic cells and weakly express CD8α and TCRβ mRNAs. In contrast, leukocytes in fractions A and B from virus-infected fish exhibit a high level of cytotoxic activity and strongly express CD8α and TCRβ mRNAs. In addition, mRNA expressions of CD8α and TCRβ in effector cells are upregulated by cocultivation with virus-infected target cells but not uninfected ones. The present study suggests that fish possess virus-specific cytotoxic cells with phenotype and gene expression pattern similar to those of CTLs in mammals.