Thymic stromal lymphopoietin limits primary and recall CD8+ T-cell anti-viral responses
- Laboratory of Molecular Immunology, Immunology Center, National Heart, Lung, and Blood Institute National Institutes of Health, United States
- Viral Immunology & Intravital Imaging Section, National Institute of Neurological Disorders and Stroke, National Institutes of Health, United States
Figures
Figure 1 with 2 supplements
TSLP acts directly on CD8+ T cells during primary influenza infection.
(A) TSLPR expression on influenza-specific CD8+ T cells (P14 tg) during primary influenza infection. Top panel, experimental design. Bottom panel, flow cytometric analysis. Naïve cells were gated on …
Figure 1—figure supplement 1
Thy1.1/Thy1.1 versus Thy1.1/Thy1.2 genetic background differences do not explain the different number of WT versus Crlf2-/- P14 T cells after influenza infection.
2.5 × 104 WT and Crlf2-/- P14 T cells were co-transferred into naive WT C57BL/6 mice (Thy1.2+/1.2+), and on the following day the mice were infected intranasally (i.n.) with PR8-33. In (A), the WT …
Figure 1—figure supplement 2
TSLP does not affect the transition to effector cells, cytokine secretion, or granzyme B levels during primary infection.
2.5 × 104 of WT (Thy1.1+/1.1+) and Crlf2-/- (Thy1.1+/1.2+) P14 T cells were co-transferred into naïve WT C57BL/6 mice (Thy1.2+/1.2+) and on the following day the mice were infected intranasally …
Figure 2
TSLP affects homeostasis of naïve but not memory CD8+ T cells.
(A and B) (A) Naïve P14 CD8+ T cells were purified and plated with either media or TSLP with or without plate-bound anti-CD3 and soluble anti-CD28. Cell viability was assessed by gating on CD8+ T …
Figure 3 with 1 supplement
TSLP modulates gene expression on naïve and memory CD8+ T cells in vitro.
RNA-Seq performed on sorted naïve and memory CD8+ T cells after 4 or 24 hr incubation with medium or TSLP with or without TCR stimulation. Shown are the number of differentially expressed genes (FC >…
Figure 3—figure supplement 1
Gating strategy for sorting memory CD8+ T cells for in vitro stimulation.
Shown is forward versus side scatter (FSC versus SSC) to exclude debris (first plot), then FSC-A and FSC-H analysis as well as SSC-H and SCC-W were performed to exclude doublets (second and third …
Figure 4 with 2 supplements
TSLP has a direct inhibitory effect on secondary CD8+ T-cell responses during influenza infection.
(A) Left panel, experimental design for (B–G), where P14 T cells were isolated from P14 WT (Thy1.1+/1.1+) and P14 Crlf2-/- (Thy1.1+/1.2+) mice and then separately injected into naïve WT mice. Each …
Figure 4—figure supplement 1
IFN-γ and TNF-α expression in WT versus Crlf2-/- mice during secondary infection.
Cells were re-stimulated with gp33 peptide in the presence of monensin and brefeldin A for 5 hr, and production of IFN-γ and TNF-α was assessed by flow cytometry. (A–C) Shown are the percentage of …
Figure 4—figure supplement 2
Gating strategy for sorting WT and Crlf2-/-CD8+ T cells for RNA-sequencing.
Shown is forward versus side scatter (FSC versus SSC) to exclude debris (first plot), then FSC-A and FSC-H analysis was performed to exclude doublets (second plot). Then dead cells were eliminated …
Figure 5 with 1 supplement
Direct actions of TSLP on virus-specific CD8+ T cells during primary and recall LCMV infection.
(A) Schematic of LCMV infection experiment. 103 of WT (Thy1.1+/1.1+) and Crlf2-/- (Thy1.1+/1.2+) P14 naïve T cells were co-transferred into naïve WT C57BL/6 mice (Thy1.2+/1.2+) and on the following …
Figure 5—figure supplement 1
TSLP protein expression during acute LCMV infection.
Additional files
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Supplementary file 1
Differentially expressed genes from sorted naïve and memory CD8+ T cells 4 or 24 hr after incubation with medium or TSLP, without TCR stimulation.
Related to Figure 3A.
- https://cdn.elifesciences.org/articles/61912/elife-61912-supp1-v1.xlsx
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Supplementary file 2
Differentially expressed genes from sorted naïve and memory CD8+ T cells 4 or 24 hr after incubation with medium or TSLP, with TCR stimulation.
Related to Figure 3B.
- https://cdn.elifesciences.org/articles/61912/elife-61912-supp2-v1.xlsx
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Supplementary file 3
RNA-Seq comparison between naïve and memory CD8+ T cells 24 hr after incubation with TSLP, without TCR stimulation.
Related to Figure 3C–E.
- https://cdn.elifesciences.org/articles/61912/elife-61912-supp3-v1.xlsx
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Supplementary file 4
Differentially expressed genes of WT versus Crlf2-/- mice, from CD8+ T cell recall responses at day eight after secondary infection.
Related to Figure 4E–G.
- https://cdn.elifesciences.org/articles/61912/elife-61912-supp4-v1.xlsx
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Transparent reporting form
- https://cdn.elifesciences.org/articles/61912/elife-61912-transrepform-v1.pdf
