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| Status |
Public on May 01, 2021 |
| Title |
Single-Cell RNA Sequencing of Thymic KN6 γδ T-Cells Modulated by TCR Signal Strength and Notch Signaling |
| Organism |
Mus musculus |
| Experiment type |
Expression profiling by high throughput sequencing
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| Summary |
γδ T-cells form an integral arm of the immune system through their rapid and potent effector functions, and are critical players during both protective and destructive immunity. However, the factors that dictate γδ T-cell functional programming in vivo remain to be fully elucidated. Here, we employed RBPJ-inducible and KN6-transgenic mice to assess the roles of ontogenic timing, T-cell receptor (TCR) signal strength, and Notch signaling in the generation of γδ T-cell functional subsets in vivo. We found skewed generation of Vγ1+ cells toward the PLZF+ γδ T-cell lineage at the fetal stage. Similarly, generation of interleukin (IL)-17 producing γδ T-cells was favored during, although not exclusive to, the fetal stage. Strong TCR signals, in conjunction with Notch, were necessary for the generation of IL-4 producing γδ T-cells. Conversely, weak TCR signals were amenable for the generation of IL-17 producing γδ T-cells, which was Notch-independent. Additionally and surprisingly, Notch signaling was also dispensable for peripheral γδ T-cell IL-17 production. Thus, our results precisely defined the roles of ontogenic timing, TCR signal strength, and Notch signaling in γδ T-cell functional programming in vivo.
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| Overall design |
CD3+Vγ4+ KN6 cells were sorted from thymi using BD FACSAria Fusion. Library construction was done using 10x Genomics Chromium Controller v3. Single-cell RNA sequencing was performed using Illumina NovaSeq 6000. Raw data were aligned to GRCm38, and raw read counts were obtained using Cell Ranger version 5.0.
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| Contributor(s) |
Chen EL, Lee CR, Thompson PK, Wiest DL, Anderson MK, Zúñiga-Pflücker JC |
| Citation(s) |
34107257 |
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| Submission date |
Feb 01, 2021 |
| Last update date |
Jun 17, 2021 |
| Contact name |
Juan Carlos Zuniga-Pflucker |
| E-mail(s) |
[email protected]
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| Organization name |
University of Toronto
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| Department |
Immunology
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| Street address |
2075 Bayview Avenue
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| City |
Toronto |
| State/province |
Ontario |
| ZIP/Postal code |
M4N 3M5 |
| Country |
Canada |
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| Platforms (1) |
| GPL24247 |
Illumina NovaSeq 6000 (Mus musculus) |
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| Samples (6)
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| Relations |
| BioProject |
PRJNA698570 |
| SRA |
SRP304123 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE165908_RAW.tar |
193.7 Mb |
(http)(custom) |
TAR (of MTX, TSV) |
SRA Run Selector |
| Raw data are available in SRA |
| Processed data provided as supplementary file |
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