Telomerase RNA component knockout exacerbates Staphylococcus aureus pneumonia by extensive inflammation and dysfunction of T cells
- Institute of Medical Microbiology, Jena University Hospital, Germany
- Else Kröner Graduate School for Medical Students 'JSAM', Jena University Hospital, Germany
- Center for Electron Microscopy, Jena University Hospital, Germany
Figures
Figure 1 with 1 supplement
Tercko/ko mice exhibit more severe pneumonia with increased mortality and a higher bacterial load.
(A) Clinical score of non-infected and infected young wild-type (WT) (n=10, 5 non-infected and 5 infected), old WT (n=10, 5 non-infected and 5 infected), and Tercko/ko (n=8, 3 non-infected and 5 …
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Figure 1—source data 1
Numeric mice data for Figure 1A–D and Figure 1—figure supplement 1A–E.
- https://cdn.elifesciences.org/articles/100433/elife-100433-fig1-data1-v1.xlsx
Figure 1—figure supplement 1
Tercko/ko mice with systemic infection display more severe pneumonia with bacteremia and increased mortality.
(A) Clinical score of infected Tercko/ko mice at the time of infection (0 dpi) and 24 hr post infection (hpi) (1 dpi). The mice were divided into three groups according to their clinical score …
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Figure 1—figure supplement 1—source data 1
Numeric data for Figure 1—figure supplement 1F.
- https://cdn.elifesciences.org/articles/100433/elife-100433-fig1-figsupp1-data1-v1.xlsx
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Figure 1—figure supplement 1—source data 2
Numeric data for Figure 1—figure supplement 1H.
- https://cdn.elifesciences.org/articles/100433/elife-100433-fig1-figsupp1-data2-v1.xlsx
Figure 2 with 1 supplement
Lungs of Tercko/ko mice display excessive inflammation and NLRP3 inflammasome activation.
(A) Pro-inflammatory cytokines measured in lung homogenates of non-infected and infected young wild-type (WT) (each n=3), old WT (each n=5), and Tercko/ko mice (non-infected: n=3, with systemic …
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Figure 2—source code 1
R code for heatmap construction.
Source Code 1 was used for the construction of all following heatmaps. Labels, title, Source Data file, and number of columns in Source Data file were adjusted for each heatmap.
- https://cdn.elifesciences.org/articles/100433/elife-100433-fig2-code1-v1.zip
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Figure 2—source code 2
R code for KEGG enrichment analysis dot plot construction.
Source Code 2 was used for all following KEGG enrichment analysis dot plot construction. Title and Source Data file were adjusted for each KEGG dot plot.
- https://cdn.elifesciences.org/articles/100433/elife-100433-fig2-code2-v1.zip
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Figure 2—source data 1
Numeric data for Figure 2A.
Table of cytokine concentrations measured.
- https://cdn.elifesciences.org/articles/100433/elife-100433-fig2-data1-v1.xlsx
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Figure 2—source data 2
Numeric data for Figure 2B.
Table of fragments per kilobase of transcript per million fragments mapped (fpkms) for all genes measured across the different mice cohorts.
- https://cdn.elifesciences.org/articles/100433/elife-100433-fig2-data2-v1.xlsx
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Figure 2—source data 3
Numeric data for Figure 2C.
List of relevant differentially expressed genes (DEGs).
- https://cdn.elifesciences.org/articles/100433/elife-100433-fig2-data3-v1.xlsx
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Figure 2—source data 4
Numeric data for Figure 2D.
List of relevant significantly enriched KEGG pathways.
- https://cdn.elifesciences.org/articles/100433/elife-100433-fig2-data4-v1.xlsx
Figure 2—figure supplement 1
Several pathways relevant for inflammation and adaptive and innate immune response are differentially expressed in infected Tercko/ko mice.
Heatmaps of differentially expressed genes (DEGs) related to the NLRP3 inflammasome pathway (A), macrophage marker (C), chemokine signaling pathway (D), T helper cell marker (E), or T cell receptor …
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Figure 2—figure supplement 1—source data 1
Numeric data for Figure 2—figure supplement 1A.
List of relevant differentially expressed genes (DEGs).
- https://cdn.elifesciences.org/articles/100433/elife-100433-fig2-figsupp1-data1-v1.xlsx
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Figure 2—figure supplement 1—source data 2
Numeric data for Figure 2—figure supplement 1C.
List of relevant differentially expressed genes (DEGs).
- https://cdn.elifesciences.org/articles/100433/elife-100433-fig2-figsupp1-data2-v1.xlsx
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Figure 2—figure supplement 1—source data 3
Numeric data for Figure 2—figure supplement 1D.
List of relevant differentially expressed genes (DEGs).
- https://cdn.elifesciences.org/articles/100433/elife-100433-fig2-figsupp1-data3-v1.xlsx
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Figure 2—figure supplement 1—source data 4
Numeric data for Figure 2—figure supplement 1E.
List of relevant differentially expressed genes (DEGs).
- https://cdn.elifesciences.org/articles/100433/elife-100433-fig2-figsupp1-data4-v1.xlsx
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Figure 2—figure supplement 1—source data 5
Numeric data for Figure 2—figure supplement 1F.
List of relevant differentially expressed genes (DEGs).
- https://cdn.elifesciences.org/articles/100433/elife-100433-fig2-figsupp1-data5-v1.xlsx
Figure 3
Elevated expression of pro-inflammatory M1 macrophages markers and chemokines are present in the lungs of infected Tercko/ko mice.
(A) Colorized scanning electron microscopy (SEM) picture of an alveolar macrophage (AM) isolated from young wild-type (WT) mice phagocytosing S. aureus. Heatmap of differentially expressed …
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Figure 3—source data 1
Numeric data for Figure 3B.
List of relevant differentially expressed genes (DEGs).
- https://cdn.elifesciences.org/articles/100433/elife-100433-fig3-data1-v1.xlsx
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Figure 3—source data 2
Numeric data for Figure 3C.
List of relevant differentially expressed genes (DEGs).
- https://cdn.elifesciences.org/articles/100433/elife-100433-fig3-data2-v1.xlsx
Figure 4 with 1 supplement
T cell receptor (TCR) in infected Tercko/ko mice is partially downregulated.
Heatmap of differentially expressed T helper cell markers (A) and differentially expressed genes (DEGs) belonging to the TCR signaling pathway (C). Red indicates upregulation, and blue indicates …
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Figure 4—source data 1
Numeric data for Figure 4A.
List of relevant differentially expressed genes (DEGs).
- https://cdn.elifesciences.org/articles/100433/elife-100433-fig4-data1-v1.xlsx
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Figure 4—source data 2
Numeric data for Figure 4B.
Percentages of live CD3+ cells.
- https://cdn.elifesciences.org/articles/100433/elife-100433-fig4-data2-v1.xlsx
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Figure 4—source data 3
Numeric data for Figure 4C.
List of relevant differentially expressed genes (DEGs).
- https://cdn.elifesciences.org/articles/100433/elife-100433-fig4-data3-v1.xlsx
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Figure 4—source data 4
Numeric data for Figure 4D–F.
Fragments per kilobase of transcript per million fragments mapped (fpkms) of the different mice cohorts for CD247, PDCD1, and CD4.
- https://cdn.elifesciences.org/articles/100433/elife-100433-fig4-data4-v1.xlsx
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Figure 4—source data 5
List of all fragments per kilobase of transcript per million fragments mapped (fpkms) for each individual mouse sequenced.
- https://cdn.elifesciences.org/articles/100433/elife-100433-fig4-data5-v1.xlsx
Figure 4—figure supplement 1
Frequency of different immune cell populations in the spleen of young and old WT as well as Tercko/ko mice.
(A) Exemplary gating strategy for the immunophenotyping of cells from murine spleen. First, lymphocytes were selected, then duplicates and dead cells were excluded (APC-Cy7 negative). To avoid …
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Figure 4—figure supplement 1—source data 1
Numeric data for Figure 4—figure supplement 1B–F.
- https://cdn.elifesciences.org/articles/100433/elife-100433-fig4-figsupp1-data1-v1.xlsx
Figure 5 with 1 supplement
T cells of Tercko/ko mice do not adequately react to bacterial challenge.
(A) Schematic representation of experiment. Murine T cells and alveolar macrophages (AMs) were isolated, co-cultured, and stimulated with heat-killed S. aureus with a multiplicity of infection 5 …
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Figure 5—source data 1
Numeric data for Figure 5D.
One replicate of the group young wild-type (WT) T cell+alveolar macrophage (AM)+ S. aureus had to be excluded due to a staining error for the displayed surface marker.
- https://cdn.elifesciences.org/articles/100433/elife-100433-fig5-data1-v1.xlsx
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Figure 5—source data 2
Numeric data for Figure 5E.
- https://cdn.elifesciences.org/articles/100433/elife-100433-fig5-data2-v1.xlsx
Figure 5—figure supplement 1
Altered immune response to infection of Tercko/ko mice relative to control cohorts.
(A) KEGG pathway enrichment analysis comparing infected Tercko/ko vs. infected old wild-type (WT), infected young WT vs. non-infected young WT, and infected Tercko/ko vs. non-infected Tercko/ko …
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Figure 5—figure supplement 1—source data 1
Numeric data for Figure 5—figure supplement 1A.
List of significantly enriched KEGG pathways of Tercko/ko S. aureus vs. old wild-type (WT) S. aureus.
- https://cdn.elifesciences.org/articles/100433/elife-100433-fig5-figsupp1-data1-v1.xlsx
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Figure 5—figure supplement 1—source data 2
Numeric data for Figure 5—figure supplement 1A.
List of significantly enriched KEGG pathways of young wild-type (WT) S. aureus vs. young WT mock.
- https://cdn.elifesciences.org/articles/100433/elife-100433-fig5-figsupp1-data2-v1.xlsx
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Figure 5—figure supplement 1—source data 3
Numeric data for Figure 5—figure supplement 1A.
List of significantly enriched KEGG pathways of Tercko/ko S. aureus vs. Tercko/ko mock.
- https://cdn.elifesciences.org/articles/100433/elife-100433-fig5-figsupp1-data3-v1.xlsx
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Figure 5—figure supplement 1—source data 4
Numeric data for Figure 5—figure supplement 1C.
- https://cdn.elifesciences.org/articles/100433/elife-100433-fig5-figsupp1-data4-v1.xlsx
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Figure 5—figure supplement 1—source data 5
Numeric data for Figure 5—figure supplement 1D.
List of relevant differentially expressed genes (DEGs).
- https://cdn.elifesciences.org/articles/100433/elife-100433-fig5-figsupp1-data5-v1.xlsx
Figure 6 with 1 supplement
Graphical summary.
Loss of Terc leads to an exaggerated inflammatory response and inflammasome activation upon S. aureus infection. This overactive inflammation disrupts T cell function, thereby impairing the immune …
Figure 6—figure supplement 1
Overview of the methods used in this publication.
Created with BioRender.com.
Additional files
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MDAR checklist
- https://cdn.elifesciences.org/articles/100433/elife-100433-mdarchecklist1-v1.docx
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Supplementary file 1
Overview of RNA concentrations measured with Nanodrop and integrity values measured with the Agilent Bioanalyzer.
- https://cdn.elifesciences.org/articles/100433/elife-100433-supp1-v1.xlsx
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Supplementary file 2
Data quality summary of each sample sequenced.
- https://cdn.elifesciences.org/articles/100433/elife-100433-supp2-v1.xlsx
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Supplementary file 3
DEGs used to construct the individual heatmaps.
- https://cdn.elifesciences.org/articles/100433/elife-100433-supp3-v1.xlsx
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Supplementary file 4
Exemplary results of genotyping of Tercko/ko mice used for breeding.
- https://cdn.elifesciences.org/articles/100433/elife-100433-supp4-v1.pptx
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Source data 1
Zip folder containing the original gel image.
- https://cdn.elifesciences.org/articles/100433/elife-100433-data1-v1.zip
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Source data 2
Zip folder containing a PDF file of the original gel with all bands and samples labeled.
- https://cdn.elifesciences.org/articles/100433/elife-100433-data2-v1.zip
