In vitro generation of viral-antigen dependent cytotoxic T-cells from ginbuna crucian carp, Carassius auratus langsdorfii
In vitro generation of viral-antigen dependent cytotoxic T-cells from ginbuna crucian carp, Carassius auratus langsdorfii
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DOI：
10.1016/j.virol.2009.04.008
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发表时间：
2009-06-01
期刊：
影响因子：
3.7
通讯作者：
Nakao, Miki
中科院分区：
文献类型：
--
作者：
Somamoto, Tomonori;Okamoto, Nobuaki;Nakao, Miki
很少有人知道抗原特异性T细胞反应的病毒在硬骨鱼由于缺乏一个合适的实验系统，使用近交系或克隆动物。在本研究中，我们已经成功地诱导在体外产生的病毒特异性细胞毒性T细胞（CTL）从同基因银鲫。从鲫鱼造血坏死病毒（CHNV）感染的鱼的反应细胞（主要是淋巴细胞）能够增殖后，在体外刺激CHNV感染的同基因刺激细胞（主要是淋巴细胞和巨噬细胞）。在体外刺激后8天和12天收集的效应细胞有效地裂解CHNV感染的同系细胞，但不裂解CHNV感染的同种异体细胞或不同病毒（伊娃）感染的同系细胞。此外，原位杂交分析表明，一些效应细胞结合CHNV感染的目标是TCR β或CD8 α阳性。这些结果提供的证据表明，在体外产生的硬骨鱼效应细胞对应于病毒特异性CTL，它们识别病毒感染的靶细胞在哺乳动物的类似方式。(c)2009 Elsevier Inc. All rights reserved.
Little is known about antigen-specific T-cell responses to viruses in teleosts due to a lack of a suitable experimental system using inbred or clonal animals. In the present study we have successfully induced an in vitro generation of virus-specific cytotoxic T-cells (CTLs) from isogeneic ginbuna crucian carp. Responder cells (primarily lymphocytes) from crucian carp haematopoietic necrosis virus (CHNV)-infected fish were capable of proliferating after stimulation in vitro with CHNV-infected syngeneic stimulator cells (primarily lymphocytes and macrophages). The effector cells collected 8 and 12 days after the in vitro stimulation efficiently lysed CHNV-infected syngeneic cells, but not CHNV-infected allogeneic cells or different virus (EVA)-infected syngeneic cells. Furthermore, in situ hybridization analysis showed that some effector cells binding to a CHNV-infected target were TCR beta or CD8 alpha positive. These results provide evidence that the teleost effector cells generated in vitro correspond to virus-specific CTL and they recognize virus-infected target cells in a similar manner of mammalian counterparts. (c) 2009 Elsevier Inc. All rights reserved.