TY - JOUR
T1 - Isolation and Characterization of Salmonid CD4+ T Cells
AU - Maisey, Kevin
AU - Montero, Ruth
AU - Corripio-Miyar, Yolanda
AU - Toro-Ascuy, Daniela
AU - Valenzuela, Beatriz
AU - Reyes-Cerpa, Sebastián
AU - Sandino, Ana María
AU - Zou, Jun
AU - Wang, Tiehui
AU - Secombes, Christopher J
AU - Imarai, Mónica
N1 - This work was supported by Fondo Nacional de Investigacio´n Cientı´fica y Tecnolo´gica Grant 1130882 and by Innova-Chile Grants 07CN13-PBT90 and 09MCSS-6698. K.M. was supported by a Comisio´n Nacional de Investigacio´n Cientı´fica y Tecnolo´gica Fellowship and by Comisio´n Nacional de Investigacio´n Cientı´fica y Tecnolo´gica, Gastos Operacionales Grant 21110476. Y.C.-M. was supported by UK National Centre for the Replacement, Refinement and Reduction of Animals in Research Grant 86682.
PY - 2016/5
Y1 - 2016/5
N2 - 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.
AB - 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.
U2 - 10.4049/jimmunol.1500439
DO - 10.4049/jimmunol.1500439
M3 - Article
C2 - 27053758
SN - 0022-1767
VL - 196
SP - 4150
EP - 4163
JO - The Journal of Immunology
JF - The Journal of Immunology
IS - 10
ER -