title: Isolation and Characterization of Salmonid CD4+ T Cells.
abstract: This study reports the isolation and functional characterization of rainbow trout (Oncorhynchus mykiss) CD4-1(+) T cells and the establishment of an IL-15-dependent CD4-1(+) T cell line. By using Abs specific for CD4-1 and CD3ε it was possible to isolate the double-positive T cells in spleen and head kidney. The morphology and the presence of transcripts for T cell markers in the sorted CD4-1(+)CD3ε(+) cells were studied next. Cells were found to express TCRα, TCRβ, CD152 (CTLA-4), CD154 (CD40L), T-bet, GATA-3, and STAT-1. The sorted CD4-1(+) T cells also had a distinctive functional attribute of mammalian T lymphocytes, namely they could undergo Ag-specific proliferation, using OVA as a model Ag. The OVA-stimulated cells showed increased expression of several cytokines, including IFN-γ1, IL-4/13A, IL-15, IL-17D, IL-10, and TGF-β1, perhaps indicating that T cell proliferation led to differentiation into distinct effector phenotypes. Using IL-15 as a growth factor, we have selected a lymphoid cell line derived from rainbow trout head kidney cells. The morphology, cell surface expression of CD4-1, and the presence of transcripts of T cell cytokines and transcription factors indicated that this is a CD4-1(+) T cell line. To our knowledge, this is the first demonstration of the presence of CD4-1(+)CD3ε(+) T cells in salmonids. As in mammals, CD4-1(+) T cells may be the master regulators of immune responses in fish, and therefore these findings and the new model T cell line developed will contribute to a greater understanding of T cell function and immune responses in teleost fish.
authors.affiliation: Laboratorio de Inmunología, Centro de Biotecnología Acuícola, Departamento de Biología, Facultad de Química y Biología, Universidad de Santiago de Chile, Alameda 3363, Santiago, Chile;
authors.forename: Kevin
authors.initials: K
authors.lastname: Maisey
authors.affiliation: Laboratorio de Inmunología, Centro de Biotecnología Acuícola, Departamento de Biología, Facultad de Química y Biología, Universidad de Santiago de Chile, Alameda 3363, Santiago, Chile;
authors.forename: Ruth
authors.initials: R
authors.lastname: Montero
authors.affiliation: Scottish Fish Immunology Research Centre, School of Biological Sciences, University of Aberdeen, Aberdeen AB24 2TZ, United Kingdom; and.
authors.forename: Yolanda
authors.initials: Y
authors.lastname: Corripio-Miyar
authors.affiliation: Laboratorio de Inmunología, Centro de Biotecnología Acuícola, Departamento de Biología, Facultad de Química y Biología, Universidad de Santiago de Chile, Alameda 3363, Santiago, Chile;
authors.forename: Daniela
authors.identifier: 0000-0002-2563-9711
authors.initials: D
authors.lastname: Toro-Ascuy
authors.affiliation: Laboratorio de Inmunología, Centro de Biotecnología Acuícola, Departamento de Biología, Facultad de Química y Biología, Universidad de Santiago de Chile, Alameda 3363, Santiago, Chile;
authors.forename: Beatriz
authors.initials: B
authors.lastname: Valenzuela
authors.affiliation: Laboratorio de Inmunología, Centro de Biotecnología Acuícola, Departamento de Biología, Facultad de Química y Biología, Universidad de Santiago de Chile, Alameda 3363, Santiago, Chile;
authors.forename: Sebastián
authors.initials: S
authors.lastname: Reyes-Cerpa
authors.affiliation: Laboratorio de Virología, Centro de Biotecnología Acuícola, Departamento de Biología, Facultad de Química y Biología, Universidad de Santiago de Chile, Alameda 3363, Santiago, Chile.
authors.forename: Ana María
authors.initials: AM
authors.lastname: Sandino
authors.affiliation: Scottish Fish Immunology Research Centre, School of Biological Sciences, University of Aberdeen, Aberdeen AB24 2TZ, United Kingdom; and.
authors.forename: Jun
authors.initials: J
authors.lastname: Zou
authors.affiliation: Scottish Fish Immunology Research Centre, School of Biological Sciences, University of Aberdeen, Aberdeen AB24 2TZ, United Kingdom; and.
authors.forename: Tiehui
authors.identifier: 0000-0002-1148-8959
authors.initials: T
authors.lastname: Wang
authors.affiliation: Scottish Fish Immunology Research Centre, School of Biological Sciences, University of Aberdeen, Aberdeen AB24 2TZ, United Kingdom; and.
authors.forename: Christopher J
authors.identifier: 0000-0001-8781-176X
authors.initials: CJ
authors.lastname: Secombes
authors.affiliation: Laboratorio de Inmunología, Centro de Biotecnología Acuícola, Departamento de Biología, Facultad de Química y Biología, Universidad de Santiago de Chile, Alameda 3363, Santiago, Chile; monica.imarai@usach.cl.
authors.forename: Mónica
authors.identifier: 0000-0001-8767-9180
authors.initials: M
authors.lastname: Imarai
mesh_terms: D000818:Animals; D017252:CD3 Complex / Q000276:immunology / Q000378:metabolism; D015496:CD4-Positive T-Lymphocytes / Q000502:physiology*; D018929:Cell Culture Techniques; D002460:Cell Line; D049109:Cell Proliferation; D029941:Fish Proteins / Q000235:genetics / Q000378:metabolism; D050990:GATA3 Transcription Factor / Q000235:genetics / Q000378:metabolism; D060226:Head Kidney / Q000276:immunology*; D019409:Interleukin-15 / Q000378:metabolism; D008213:Lymphocyte Activation; D008322:Mammals / Q000276:immunology; D017686:Oncorhynchus mykiss / Q000276:immunology*; D010802:Phylogeny; D011948:Receptors, Antigen, T-Cell / Q000378:metabolism; D050794:STAT1 Transcription Factor / Q000235:genetics / Q000378:metabolism; D013154:Spleen / Q000276:immunology*; D020825:T-Box Domain Proteins / Q000235:genetics / Q000378:metabolism; D000098815:T-bet Transcription Factor
chemical_list: D017252:CD3 Complex; D029941:Fish Proteins; D050990:GATA3 Transcription Factor; D019409:Interleukin-15; D011948:Receptors, Antigen, T-Cell; D050794:STAT1 Transcription Factor; D020825:T-Box Domain Proteins; D000098815:T-bet Transcription Factor