DataSheet_3_Leptin deficiency in CD8+ T cells ameliorates non-segmental vitiligo by reducing interferon-γ and Granzyme B.zip

BackgroundVitiligo is an autoimmune skin disease mainly mediated by CD8+ T cells, which affects about 0.1%-2% population of the world. Leptin plays a critical role in regulating the activation of CD8+ T cells. However, the effect of Leptin on vitiligo remains unclear.ObjectivesTo explore the effect of leptin on CD8+ T cells and its influence on vitiligo.MethodsRNA sequencing and Quantitative Real-time PCR (RT-qPCR) were used to explore the differentially expressed genes. Immunofluorescence staining was performed on skin lesions. Leptin in serum was detected by enzyme linked immunosorbent assay (ELISA). The peripheral blood mononuclear cells were detected by flow cytometry after leptin stimulation for 72 hours. A vitiligo model was established by monobenzone on Leptin KO mice.Results557 differentially expressed genes were found, including 154 up-regulated and 403 down-regulated genes. Lipid metabolism pathways showed a close relationship to the pathogenesis of vitiligo, especially the PPAR signaling pathway. RT-qPCR (p = 0.013) and immunofluorescence staining (p = 0.0053) verified that LEPR expressed significantly higher in vitiligo. The serum leptin level of vitiligo patients was significantly lower than that of healthy controls (p = 0.0245). The interferon-γ subset of CD8+LEPR+ T cells from vitiligo patients was significantly higher (p = 0.0189). The protein level of interferon-γ was significantly increased after leptin stimulation in vitro (p = 0.0217). In mice, Leptin deficiency resulted in less severe hair depigmentation. Leptin deficiency also resulted in significantly lower expressed vitiligo-related genes, such as Cxcl9 (p = 0.0497), Gzmb (p ConclusionLeptin could promote the progression of vitiligo by enhancing the cytotoxic function of CD8+ T cells. Leptin may become a new target for vitiligo treatment.</p

DataSheet_1_Leptin deficiency in CD8+ T cells ameliorates non-segmental vitiligo by reducing interferon-γ and Granzyme B.xls

BackgroundVitiligo is an autoimmune skin disease mainly mediated by CD8+ T cells, which affects about 0.1%-2% population of the world. Leptin plays a critical role in regulating the activation of CD8+ T cells. However, the effect of Leptin on vitiligo remains unclear.ObjectivesTo explore the effect of leptin on CD8+ T cells and its influence on vitiligo.MethodsRNA sequencing and Quantitative Real-time PCR (RT-qPCR) were used to explore the differentially expressed genes. Immunofluorescence staining was performed on skin lesions. Leptin in serum was detected by enzyme linked immunosorbent assay (ELISA). The peripheral blood mononuclear cells were detected by flow cytometry after leptin stimulation for 72 hours. A vitiligo model was established by monobenzone on Leptin KO mice.Results557 differentially expressed genes were found, including 154 up-regulated and 403 down-regulated genes. Lipid metabolism pathways showed a close relationship to the pathogenesis of vitiligo, especially the PPAR signaling pathway. RT-qPCR (p = 0.013) and immunofluorescence staining (p = 0.0053) verified that LEPR expressed significantly higher in vitiligo. The serum leptin level of vitiligo patients was significantly lower than that of healthy controls (p = 0.0245). The interferon-γ subset of CD8+LEPR+ T cells from vitiligo patients was significantly higher (p = 0.0189). The protein level of interferon-γ was significantly increased after leptin stimulation in vitro (p = 0.0217). In mice, Leptin deficiency resulted in less severe hair depigmentation. Leptin deficiency also resulted in significantly lower expressed vitiligo-related genes, such as Cxcl9 (p = 0.0497), Gzmb (p ConclusionLeptin could promote the progression of vitiligo by enhancing the cytotoxic function of CD8+ T cells. Leptin may become a new target for vitiligo treatment.</p

DataSheet_4_Leptin deficiency in CD8+ T cells ameliorates non-segmental vitiligo by reducing interferon-γ and Granzyme B.docx

BackgroundVitiligo is an autoimmune skin disease mainly mediated by CD8+ T cells, which affects about 0.1%-2% population of the world. Leptin plays a critical role in regulating the activation of CD8+ T cells. However, the effect of Leptin on vitiligo remains unclear.ObjectivesTo explore the effect of leptin on CD8+ T cells and its influence on vitiligo.MethodsRNA sequencing and Quantitative Real-time PCR (RT-qPCR) were used to explore the differentially expressed genes. Immunofluorescence staining was performed on skin lesions. Leptin in serum was detected by enzyme linked immunosorbent assay (ELISA). The peripheral blood mononuclear cells were detected by flow cytometry after leptin stimulation for 72 hours. A vitiligo model was established by monobenzone on Leptin KO mice.Results557 differentially expressed genes were found, including 154 up-regulated and 403 down-regulated genes. Lipid metabolism pathways showed a close relationship to the pathogenesis of vitiligo, especially the PPAR signaling pathway. RT-qPCR (p = 0.013) and immunofluorescence staining (p = 0.0053) verified that LEPR expressed significantly higher in vitiligo. The serum leptin level of vitiligo patients was significantly lower than that of healthy controls (p = 0.0245). The interferon-γ subset of CD8+LEPR+ T cells from vitiligo patients was significantly higher (p = 0.0189). The protein level of interferon-γ was significantly increased after leptin stimulation in vitro (p = 0.0217). In mice, Leptin deficiency resulted in less severe hair depigmentation. Leptin deficiency also resulted in significantly lower expressed vitiligo-related genes, such as Cxcl9 (p = 0.0497), Gzmb (p ConclusionLeptin could promote the progression of vitiligo by enhancing the cytotoxic function of CD8+ T cells. Leptin may become a new target for vitiligo treatment.</p

DataSheet_2_Leptin deficiency in CD8+ T cells ameliorates non-segmental vitiligo by reducing interferon-γ and Granzyme B.xls

BackgroundVitiligo is an autoimmune skin disease mainly mediated by CD8+ T cells, which affects about 0.1%-2% population of the world. Leptin plays a critical role in regulating the activation of CD8+ T cells. However, the effect of Leptin on vitiligo remains unclear.ObjectivesTo explore the effect of leptin on CD8+ T cells and its influence on vitiligo.MethodsRNA sequencing and Quantitative Real-time PCR (RT-qPCR) were used to explore the differentially expressed genes. Immunofluorescence staining was performed on skin lesions. Leptin in serum was detected by enzyme linked immunosorbent assay (ELISA). The peripheral blood mononuclear cells were detected by flow cytometry after leptin stimulation for 72 hours. A vitiligo model was established by monobenzone on Leptin KO mice.Results557 differentially expressed genes were found, including 154 up-regulated and 403 down-regulated genes. Lipid metabolism pathways showed a close relationship to the pathogenesis of vitiligo, especially the PPAR signaling pathway. RT-qPCR (p = 0.013) and immunofluorescence staining (p = 0.0053) verified that LEPR expressed significantly higher in vitiligo. The serum leptin level of vitiligo patients was significantly lower than that of healthy controls (p = 0.0245). The interferon-γ subset of CD8+LEPR+ T cells from vitiligo patients was significantly higher (p = 0.0189). The protein level of interferon-γ was significantly increased after leptin stimulation in vitro (p = 0.0217). In mice, Leptin deficiency resulted in less severe hair depigmentation. Leptin deficiency also resulted in significantly lower expressed vitiligo-related genes, such as Cxcl9 (p = 0.0497), Gzmb (p ConclusionLeptin could promote the progression of vitiligo by enhancing the cytotoxic function of CD8+ T cells. Leptin may become a new target for vitiligo treatment.</p

Supplementary Figure – Comprehensive analysis of epigenetic modifications and immune-cell infiltration in tissues from SLE patients

Supplementary Figure 1 Identification of DEGs and enrichment of DEGs (A)Volcano plot of datasets in normal and SLE samples from GSE20864. Blue plots represent expressions of genes with P < 0.05 and log2FC < −0.8. Red plots represent expressions of genes mRNA with P  0.8. Grey plots represent genes expressed mRNA normally. The X-axis means the log2 of fold change in expressions of genes between primary and metastatic samples. The Y-axis means the −log10 of the P value of each gene. (B) Volcano plot of datasets in normal and SLE samples from GSE112087. (C) Volcano plot of datasets in normal and SLE samples from GSE122459. (D) Volcano plot of datasets in normal and SLE samples from GSE144390. (E) Enrichment of DEGs in 4 datasets. Supplementary Figure 2 Correlation analysis of immune cells and key genes The red circle represents positive correlation, and the purple circle represents negative correlation (P Supplementary Figure 3 Identification of different immune cells in single cell analysis (A) PBMC cells from 5 normal subjects and 7 SLE patients are divided into 19 clusters. Each point represents a single cell, colored according to clusters. (B) The amount of marker genes expressed by each cluster. (C) Relative frequency of expression of each immune cell in SLE and HC Supplementary Figure 4 Epigenetic analysis of SLE (A) Enrichment of hypomethylation gene. (B) Volcano plot of datasets in normal and SLE samples from GSE37426. (C) Heatmap of DEmiRNAs between normal and SLE samples from GSE37426. (D) Volcano plot of datasets in normal and SLE samples from GSE102547. (E) Heatmap of DElncRNAs between normal and SLE samples from GSE102547. (F) Volcano plot of datasets in normal and SLE samples from GSE84655. (G) Heatmap of DEcircRNAs between normal and SLE samples from GSE84655. Supplementary Figure 5 The mRNA expression of 5 key genes in HC and autoimmune diseases The relative mRNA expression of RSAD2, OAS2, IFIT1, IFIT3, PLSCR1 in the PBMCs of HC, SLE, RA, DM, SS. Supplementary Figure 6 The protein expression of IFIT1 and RSAD2 in SLE and HC skin lesions. (A) The statistical analysis percentage of IFIT1 positive cells between SLE and HC. (B) The statistical analysis percentage of RSAD2 positive cells between SLE and HC. </div

Supplementary Tables – Comprehensive analysis of epigenetic modifications and immune-cell infiltration in tissues from SLE patients

Supplementary Table 1 Primers for RT-qPCR Supplementary Table 2 The number of patients in GEO datasets Supplementary Table 3 Marker genes of cells Supplementary Table 4 Analysis of key genes expressions in GSE185047 </p