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(A-C) scRNAseq dot plot of epithelial cells separated by (A) fungal colonization type or (B) epithelial cell type or (C) IFNγ -/- , WT, IL-17 -/- , and IFNγ OE genotype. Mice were treated with Candida spp indicated on days 0,2,4,6 and back skin was harvested on day 14. (D) Representative 3D confocal microscopy of Stat1 expression in the upper hair follicle in WT mice treated with either mock colonization or C. auris colonization on day 14. (E) Correlation between Stat1 expression, assessed via mean fluorescence index (MFI), and CD8 T cells per hair follicle. (F) Quantitation of Stat1 mean fluorescence intensity (MFI) per hair follicle in mice indicated. WT C. auris colonized mice were subdivided based on having less than or greater than 15 CD8 T-cells per hair follicle. (G) Stat1 MFI in the epithelial surface (CD49f) compared to all other skin structures (non-CD49f). (H) Stat1 MFI between the interfollicular epithelium (IFE), upper hair follicle, and lower hair follicle in mock colonized (white) and C. auris colonized (red) mice. (I,J) Schematics showing models used to test colonization and epicutaneous infection. For epicutaneous infection, intact back skin was pre-colonized with C. auris on day 0,2,4,6; on day 14, hair was removed, and the skin was abraded with sandpaper. C. auris was reapplied on day 15 and back skin was harvested 2 days later for fungal burden. For ‘Re-Colonization’, intact back skin was pre-colonized with C. auris on days 0,2,4,6 and intact back skin was re-colonized on day 14. Back skin was harvested for fungal CFU 5 days after final colonization. Graph showing fungal burden (CFU) after C. auris (I) epicutaneous infection and (J) recolonization of B6 wild type (WT), IFNγ -/- , IL-17 -/- mice, IFNγ OE mice (Yeti/+), and Ker14 ΔIFNγR1 (Ker14 Cre <t>;IFNγR1</t> Flox ). (K) IFNγ overexpressing mice ( IFNγ OE ) colonized with C. auris , at day 14 with numerous yeast forms at the hair follicle orifice. *P<0.05, ****P<0.0001 for one-way analysis of variance (ANOVA) and Tukey’s multiple comparison test (F) . In (E), statistics shown comparing C. auris <15 CD8 per hair follicle to all other groups. **P<0.01, ***P<0.001 for Kruskal-Wallis test with Dunn’s multiple comparison test (I,J) . *P<0.05, **P<0.01 student T-Test (G,H). *P<0.05, **P<0.01 Mann-Whitney Test (I,J) .
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(A-C) scRNAseq dot plot of epithelial cells separated by (A) fungal colonization type or (B) epithelial cell type or (C) IFNγ -/- , WT, IL-17 -/- , and IFNγ OE genotype. Mice were treated with Candida spp indicated on days 0,2,4,6 and back skin was harvested on day 14. (D) Representative 3D confocal microscopy of Stat1 expression in the upper hair follicle in WT mice treated with either mock colonization or C. auris colonization on day 14. (E) Correlation between Stat1 expression, assessed via mean fluorescence index (MFI), and CD8 T cells per hair follicle. (F) Quantitation of Stat1 mean fluorescence intensity (MFI) per hair follicle in mice indicated. WT C. auris colonized mice were subdivided based on having less than or greater than 15 CD8 T-cells per hair follicle. (G) Stat1 MFI in the epithelial surface (CD49f) compared to all other skin structures (non-CD49f). (H) Stat1 MFI between the interfollicular epithelium (IFE), upper hair follicle, and lower hair follicle in mock colonized (white) and C. auris colonized (red) mice. (I,J) Schematics showing models used to test colonization and epicutaneous infection. For epicutaneous infection, intact back skin was pre-colonized with C. auris on day 0,2,4,6; on day 14, hair was removed, and the skin was abraded with sandpaper. C. auris was reapplied on day 15 and back skin was harvested 2 days later for fungal burden. For ‘Re-Colonization’, intact back skin was pre-colonized with C. auris on days 0,2,4,6 and intact back skin was re-colonized on day 14. Back skin was harvested for fungal CFU 5 days after final colonization. Graph showing fungal burden (CFU) after C. auris (I) epicutaneous infection and (J) recolonization of B6 wild type (WT), IFNγ -/- , IL-17 -/- mice, IFNγ OE mice (Yeti/+), and Ker14 ΔIFNγR1 (Ker14 Cre <t>;IFNγR1</t> Flox ). (K) IFNγ overexpressing mice ( IFNγ OE ) colonized with C. auris , at day 14 with numerous yeast forms at the hair follicle orifice. *P<0.05, ****P<0.0001 for one-way analysis of variance (ANOVA) and Tukey’s multiple comparison test (F) . In (E), statistics shown comparing C. auris <15 CD8 per hair follicle to all other groups. **P<0.01, ***P<0.001 for Kruskal-Wallis test with Dunn’s multiple comparison test (I,J) . *P<0.05, **P<0.01 student T-Test (G,H). *P<0.05, **P<0.01 Mann-Whitney Test (I,J) .
C57bl 6n Ifngr1tm1 1 Rds J Mice, supplied by Jackson Laboratory, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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(A-C) scRNAseq dot plot of epithelial cells separated by (A) fungal colonization type or (B) epithelial cell type or (C) IFNγ -/- , WT, IL-17 -/- , and IFNγ OE genotype. Mice were treated with Candida spp indicated on days 0,2,4,6 and back skin was harvested on day 14. (D) Representative 3D confocal microscopy of Stat1 expression in the upper hair follicle in WT mice treated with either mock colonization or C. auris colonization on day 14. (E) Correlation between Stat1 expression, assessed via mean fluorescence index (MFI), and CD8 T cells per hair follicle. (F) Quantitation of Stat1 mean fluorescence intensity (MFI) per hair follicle in mice indicated. WT C. auris colonized mice were subdivided based on having less than or greater than 15 CD8 T-cells per hair follicle. (G) Stat1 MFI in the epithelial surface (CD49f) compared to all other skin structures (non-CD49f). (H) Stat1 MFI between the interfollicular epithelium (IFE), upper hair follicle, and lower hair follicle in mock colonized (white) and C. auris colonized (red) mice. (I,J) Schematics showing models used to test colonization and epicutaneous infection. For epicutaneous infection, intact back skin was pre-colonized with C. auris on day 0,2,4,6; on day 14, hair was removed, and the skin was abraded with sandpaper. C. auris was reapplied on day 15 and back skin was harvested 2 days later for fungal burden. For ‘Re-Colonization’, intact back skin was pre-colonized with C. auris on days 0,2,4,6 and intact back skin was re-colonized on day 14. Back skin was harvested for fungal CFU 5 days after final colonization. Graph showing fungal burden (CFU) after C. auris (I) epicutaneous infection and (J) recolonization of B6 wild type (WT), IFNγ -/- , IL-17 -/- mice, IFNγ OE mice (Yeti/+), and Ker14 ΔIFNγR1 (Ker14 Cre <t>;IFNγR1</t> Flox ). (K) IFNγ overexpressing mice ( IFNγ OE ) colonized with C. auris , at day 14 with numerous yeast forms at the hair follicle orifice. *P<0.05, ****P<0.0001 for one-way analysis of variance (ANOVA) and Tukey’s multiple comparison test (F) . In (E), statistics shown comparing C. auris <15 CD8 per hair follicle to all other groups. **P<0.01, ***P<0.001 for Kruskal-Wallis test with Dunn’s multiple comparison test (I,J) . *P<0.05, **P<0.01 student T-Test (G,H). *P<0.05, **P<0.01 Mann-Whitney Test (I,J) .
Young Adult Male C57bl 6n Mice, supplied by Jackson Laboratory, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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(A-C) scRNAseq dot plot of epithelial cells separated by (A) fungal colonization type or (B) epithelial cell type or (C) IFNγ -/- , WT, IL-17 -/- , and IFNγ OE genotype. Mice were treated with Candida spp indicated on days 0,2,4,6 and back skin was harvested on day 14. (D) Representative 3D confocal microscopy of Stat1 expression in the upper hair follicle in WT mice treated with either mock colonization or C. auris colonization on day 14. (E) Correlation between Stat1 expression, assessed via mean fluorescence index (MFI), and CD8 T cells per hair follicle. (F) Quantitation of Stat1 mean fluorescence intensity (MFI) per hair follicle in mice indicated. WT C. auris colonized mice were subdivided based on having less than or greater than 15 CD8 T-cells per hair follicle. (G) Stat1 MFI in the epithelial surface (CD49f) compared to all other skin structures (non-CD49f). (H) Stat1 MFI between the interfollicular epithelium (IFE), upper hair follicle, and lower hair follicle in mock colonized (white) and C. auris colonized (red) mice. (I,J) Schematics showing models used to test colonization and epicutaneous infection. For epicutaneous infection, intact back skin was pre-colonized with C. auris on day 0,2,4,6; on day 14, hair was removed, and the skin was abraded with sandpaper. C. auris was reapplied on day 15 and back skin was harvested 2 days later for fungal burden. For ‘Re-Colonization’, intact back skin was pre-colonized with C. auris on days 0,2,4,6 and intact back skin was re-colonized on day 14. Back skin was harvested for fungal CFU 5 days after final colonization. Graph showing fungal burden (CFU) after C. auris (I) epicutaneous infection and (J) recolonization of B6 wild type (WT), IFNγ -/- , IL-17 -/- mice, IFNγ OE mice (Yeti/+), and Ker14 ΔIFNγR1 (Ker14 Cre ;IFNγR1 Flox ). (K) IFNγ overexpressing mice ( IFNγ OE ) colonized with C. auris , at day 14 with numerous yeast forms at the hair follicle orifice. *P<0.05, ****P<0.0001 for one-way analysis of variance (ANOVA) and Tukey’s multiple comparison test (F) . In (E), statistics shown comparing C. auris <15 CD8 per hair follicle to all other groups. **P<0.01, ***P<0.001 for Kruskal-Wallis test with Dunn’s multiple comparison test (I,J) . *P<0.05, **P<0.01 student T-Test (G,H). *P<0.05, **P<0.01 Mann-Whitney Test (I,J) .

Journal: bioRxiv

Article Title: The emerging fungal pathogen Candida auris induces IFNγ to colonize mammalian hair follicles

doi: 10.1101/2025.01.15.632653

Figure Lengend Snippet: (A-C) scRNAseq dot plot of epithelial cells separated by (A) fungal colonization type or (B) epithelial cell type or (C) IFNγ -/- , WT, IL-17 -/- , and IFNγ OE genotype. Mice were treated with Candida spp indicated on days 0,2,4,6 and back skin was harvested on day 14. (D) Representative 3D confocal microscopy of Stat1 expression in the upper hair follicle in WT mice treated with either mock colonization or C. auris colonization on day 14. (E) Correlation between Stat1 expression, assessed via mean fluorescence index (MFI), and CD8 T cells per hair follicle. (F) Quantitation of Stat1 mean fluorescence intensity (MFI) per hair follicle in mice indicated. WT C. auris colonized mice were subdivided based on having less than or greater than 15 CD8 T-cells per hair follicle. (G) Stat1 MFI in the epithelial surface (CD49f) compared to all other skin structures (non-CD49f). (H) Stat1 MFI between the interfollicular epithelium (IFE), upper hair follicle, and lower hair follicle in mock colonized (white) and C. auris colonized (red) mice. (I,J) Schematics showing models used to test colonization and epicutaneous infection. For epicutaneous infection, intact back skin was pre-colonized with C. auris on day 0,2,4,6; on day 14, hair was removed, and the skin was abraded with sandpaper. C. auris was reapplied on day 15 and back skin was harvested 2 days later for fungal burden. For ‘Re-Colonization’, intact back skin was pre-colonized with C. auris on days 0,2,4,6 and intact back skin was re-colonized on day 14. Back skin was harvested for fungal CFU 5 days after final colonization. Graph showing fungal burden (CFU) after C. auris (I) epicutaneous infection and (J) recolonization of B6 wild type (WT), IFNγ -/- , IL-17 -/- mice, IFNγ OE mice (Yeti/+), and Ker14 ΔIFNγR1 (Ker14 Cre ;IFNγR1 Flox ). (K) IFNγ overexpressing mice ( IFNγ OE ) colonized with C. auris , at day 14 with numerous yeast forms at the hair follicle orifice. *P<0.05, ****P<0.0001 for one-way analysis of variance (ANOVA) and Tukey’s multiple comparison test (F) . In (E), statistics shown comparing C. auris <15 CD8 per hair follicle to all other groups. **P<0.01, ***P<0.001 for Kruskal-Wallis test with Dunn’s multiple comparison test (I,J) . *P<0.05, **P<0.01 student T-Test (G,H). *P<0.05, **P<0.01 Mann-Whitney Test (I,J) .

Article Snippet: To generate keratinocyte specific knock out of IL-17 and IFNγ signaling, we crossed Ker14 Cre mice (B6N.Cg-Tg(KRT14-cre)1Amc/J) from Jackson with Il17Ra floxed mice (B6.Cg-Il17ratm2.1Koll/J; MGI:J:237978) and IFNγR1 floxed mice (C57BL/6N-Ifngr1tm1.1Rds/J, MGI:J:206257) from Jackson Labs. To track type 1 lymphocytes we used Tbet (Tbx21)-zsGreen transgenic mice (generously provided by Jinfang Zhu, Lab of Immune System Biology, NIH).

Techniques: Confocal Microscopy, Expressing, Fluorescence, Quantitation Assay, Infection, Comparison, MANN-WHITNEY