UMMS Affiliation

Department of Pathology; Graduate School of Biomedical Sciences

Publication Date

2018-08-01

Document Type

Article

Disciplines

Amino Acids, Peptides, and Proteins | Cell Biology | Cells | Hemic and Immune Systems | Immunopathology

Abstract

Circulating naive T cells exist in a quiescent state. After TCR contact with the cognate peptide presented by APCs in secondary lymphoid structures, T cells undergo a period of rapid transcriptional changes that set the stage for fate-determining effector or memory programming. We describe a novel method to analyze TCR signaling pathway activation in nuclei isolated from primary mouse naive T cells after stimulation with natural peptide Ags. We prelabeled cells with cell tracking dye to easily distinguish CD8(+) T cell nuclei from APC nuclei by conventional flow cytometry. Using this approach, we observed clear digital activation of NFAT1 transcription factor in OT-I T cells stimulated with OVA peptide presented by bulk splenocytes. OVA concentration had discrete control over the fraction of the cells that translocated NFAT1, indicating that a distinct threshold amount of TCR signaling is required to switch on NFAT1 in naive T cells. This behavior was cell contact dependent and qualitatively more exact than the NFAT1 response in ionomycin-stimulated naive T cells. These data contribute to our understanding of the digital behavior of TCR signaling components documented in other studies and indicate how T cells might discriminate log-fold changes in Ag availability during an actual infection. Overall, these results highlight the potential of this coculture nuclei isolation protocol to address stimulation-dependent translocation of proteins in primary lymphocytes.

Rights and Permissions

Copyright © 2018 The Authors. This article is distributed under the terms of the CC BY-NC-ND 4.0 Unported license.

DOI of Published Version

10.4049/immunohorizons.1800032

Source

Immunohorizons. 2018 Aug;2(7):208-215. doi: 10.4049/immunohorizons.1800032. Link to article on publisher's site

Journal/Book/Conference Title

ImmunoHorizons

Related Resources

Link to Article in PubMed

PubMed ID

30221251

Creative Commons License

Creative Commons Attribution-Noncommercial-No Derivative Works 4.0 License
This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 4.0 License.

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