Isolation and Characterization of Salmonid CD4+ T Cells

Kevin Maisey, Ruth Montero, Yolanda Corripio-Miyar, Daniela Toro-Ascuy, Beatriz Valenzuela, Sebastián Reyes-Cerpa, Ana María Sandino, Jun Zou, Tiehui Wang, Christopher J Secombes, Mónica Imarai

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26 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)4150-4163
Number of pages14
JournalThe Journal of Immunology
Volume196
Issue number10
Early online date6 Apr 2016
DOIs
Publication statusPublished - May 2016

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    Maisey, K., Montero, R., Corripio-Miyar, Y., Toro-Ascuy, D., Valenzuela, B., Reyes-Cerpa, S., Sandino, A. M., Zou, J., Wang, T., Secombes, C. J., & Imarai, M. (2016). Isolation and Characterization of Salmonid CD4+ T Cells. The Journal of Immunology, 196(10), 4150-4163. https://doi.org/10.4049/jimmunol.1500439