title: Clonal growth of carp (Cyprinus carpio) T cells in vitro: long-term proliferation of Th2-like cells.
abstract: Carp kidney leukocytes co-cultured with a supporting cell layer resulted in proliferation of polyclonal CD4(+) αβT cells as described previously. These bulk-cultured T cells expressed transcripts for both T helper 1 cells (Th1) master regulator (T-bet) and T helper 2 cells (Th2) master regulator (GATA-3). To identify the Th subsets in bulk-cultured T cells, single cells were picked up from the bulk culture, proliferated, and characterized. The majority of the clones displayed characteristics consistent with CD4(+) αβT cell identity. These clones expressed both TCRα and TCRβ, but could not produce a TCRγδ heterodimer since they typically only expressed either TCRγ or TCRδ. These clones also expressed the TCR co-receptor genes CD4-1 or CD4-2, whereas they did not express CD8α or CD8β. In addition, GATA-3 was expressed whereas T-bet was not. Among these clones, one clone (KoThL5) continued to proliferate on the supporting cells and was successively transferred for more than 10 months and 90-100 passages. To characterize the KoThL5 cells by their cytokine production profile, they were stimulated with PHA and investigated by real-time RT-PCR. mRNA expression of Th2-related cytokine (IL-4/13B) was only enhanced in KoThL5 cells whereas both Th1-related cytokine (IFNγ) and Th2-related cytokines (IL-4/13A and IL-4/13B) were significantly enhanced in bulk-cultured T cells. Taken together, KoThL5 cells share some features with mammalian Th2 cells. This is the first study to describe in vitro cultures of teleost cell with Th2-like features. The KoThL5 cell line has considerable potential for addressing questions concerning the properties of teleost Th2 cells.
authors.affiliation: Laboratory of Fish Pathology, Department of Veterinary Medicine, Nihon University, Fujisawa, Kanagawa, Japan.
authors.forename: Takuya
authors.initials: T
authors.lastname: Yamaguchi
authors.forename: Fumihiko
authors.initials: F
authors.lastname: Katakura
authors.forename: Kazue
authors.initials: K
authors.lastname: Someya
authors.forename: Johannes Martinus
authors.initials: JM
authors.lastname: Dijkstra
authors.forename: Tadaaki
authors.initials: T
authors.lastname: Moritomo
authors.forename: Teruyuki
authors.initials: T
authors.lastname: Nakanishi
mesh_terms: D000818:Animals; D002347:Carps / Q000276:immunology*; D049109:Cell Proliferation; D016207:Cytokines / Q000378:metabolism; D050990:GATA3 Transcription Factor / Q000235:genetics / Q000378:metabolism; D007621:Karyotyping / Q000662:veterinary; D007668:Kidney / Q000166:cytology; D060888:Real-Time Polymerase Chain Reaction / Q000662:veterinary; D011948:Receptors, Antigen, T-Cell / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000662:veterinary; D020825:T-Box Domain Proteins / Q000235:genetics / Q000378:metabolism; D018418:Th2 Cells / Q000166:cytology* / Q000378:metabolism / Q000502:physiology; D000098815:T-bet Transcription Factor
chemical_list: D016207:Cytokines; D050990:GATA3 Transcription Factor; D011948:Receptors, Antigen, T-Cell; D020825:T-Box Domain Proteins; D000098815:T-bet Transcription Factor