In vitro generation of viral-antigen dependent cytotoxic T-cells from ginbuna crucian carp, Carassius auratus langsdorfii.
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 TCRbeta or CD8alpha 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.
Laboratory of Marine Biochemistry, Department of Bioscience and Biotechnology, Graduate School of Bioresource and Bioenvironmental Sciences, Kyushu University, Hakozaki, Fukuoka 812-8581, Japan. somamoto@agr.kyushu-u.ac.jp
Tomonori
T
Somamoto
Nobuaki
N
Okamoto
Teruyuki
T
Nakanishi
Mitsuru
M
Ototake
Miki
M
Nakao
D000818:Animals; D000938:Antigen-Presenting Cells / Q000276:immunology / Q000821:virology; D000956:Antigens, Viral / Q000276:immunology*; D016827:CD8 Antigens / Q000378:metabolism; D002347:Carps / Q000276:immunology* / Q000821:virology; D002460:Cell Line; D049109:Cell Proliferation; D018920:Coculture Techniques; D012333:RNA, Messenger / Q000378:metabolism; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000378:metabolism; D012209:Rhabdoviridae / Q000276:immunology*; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology* / Q000821:virology
D000956:Antigens, Viral; D016827:CD8 Antigens; D012333:RNA, Messenger; D016693:Receptors, Antigen, T-Cell, alpha-beta