Journal: Nature Communications

Article Title: Immune checkpoint inhibitor-induced severe epidermal necrolysis mediated by macrophage-derived CXCL10 and abated by TNF blockade

doi: 10.1038/s41467-024-54180-7

Figure Lengend Snippet: a Analysis of chemokine receptor–ligand pairs (CXCR3–CXCL9 and CXCR3–CXCL10) across clusters in all cell datasets shown in Fig. . The “ICI-SJS/TEN” group includes lesional BC and PBMC from 5 patients with ICI-induced SJS/TEN patients, and the “ICI-tolerant & HD” group includes PBMC from 5 ICI-tolerant patients and 6 healthy donors (HD). b The scaled and normalized gene expression levels for CXCL9 and CXCL10 in all cell datasets (related to Fig. ) was shown, whereas the violin plot and heatmap beneath was drawn from the subset of macrophage/monocyte/myeloid clusters. c The scaled and normalized expression levels of CXCR3 in all cell datasets (related to Fig. ) were shown, whereas the violin plot and heatmap beneath were drawn from the subset of CD8 + T cell clusters. ISB: ICI-SJS/TEN lesional BC; ISP: ICI-SJS/TEN PBMC; IMP: ICI-mild cADR PBMC; ITP: ICI-tolerant PBMC; HD: healthy donors PBMC. d Heatmap showing Z score–normalized averaged expression levels of the indicated cytokines/cytotoxic proteins/inflammatory proteins and its related receptor genes compared among ICI-SJS/TEN, ICI-mild cADR, and control groups. The normalized differential gene expression (DEG) levels in CD8 + T cell clusters (clusters 3–7 in Fig. ; defined by SingleR annotation) and macrophage/monocyte/myeloid clusters (clusters 9–14 in Fig. ; defined by SingleR annotation) were shown. The indicated relevant genes list is based on the significant P values for ICI-SJS/TEN lesional BC group comparing to “ICI-tolerant & HD” group, which were calculated by Wilcoxon rank-sum test. The genes associated with the TNF signaling pathway were denoted in a red color. e Scatterplot showing DEG, confirming RNA expression levels in formalin-fixed paraffin-embedded (FFPE) skin tissue samples from patients with ICI-induced SJS–TEN (ICI-SJS/TEN; n = 5) or ICI-induced lichenoid dermatitis (ICI-mild cADR; n = 7) compared with those from HD (HD; n = 8). Each dot denotes an individual gene with a Benjamini–Hochberg-adjusted P value (two-sided unpaired Mann–Whitney U test) <0.05 and average log 2 fold change (FC) > 2 in ICI-SJS/TEN samples compared with HD samples (ICI-SJS/TEN vs. HD) and in ICI-mild cADR samples compared with HD samples (ICI-mild cADR vs. HD). The blue-labeled genes represent genes that were significantly elevated in both the ICI-SJS/TEN vs. HD and ICI-mild cADR vs. HD comparisons, whereas the red-labeled genes were only significantly elevated in the ICI-SJS/TEN vs. HD comparison. f Immunofluorescence staining with anti-CD8 (green) and anti-CXCR3 (red) antibodies, and 4’,6-diamidino-2-phenylindole (DAPI; nuclear stain; blue) in skin tissues from 7 patients with ICI-SJS/TEN and 4 ICI-mild cADR and from 4 HD control participants (sample list shown in source data). The upper figures represent 200×, whereas the lower figures represent 400× magnification. g Immunofluorescence staining with anti-CXCL10 (green) and anti-CD163 (red) antibodies and DAPI (blue). The upper figures represent 200×, whereas the lower figures represent 400× magnification.

Article Snippet: The following primary antibodies were used: rabbit monoclonal anti-human CXCL10-FITC antibody (1:100 dilution, LS-C123903, LSBio), mouse monoclonal anti-human CD163 antibody (1:100 dilution, ab156769, Abcam), mouse monoclonal anti-human CD8 antibody (1:100 dilution, ab199016, Abcam), and rabbit polyclonal anti-human CXCR3 antibody (1:200 dilution, ab71864, Abcam).

Techniques: Expressing, Control, RNA Expression, Formalin-fixed Paraffin-Embedded, MANN-WHITNEY, Labeling, Comparison, Immunofluorescence, Staining

Journal: Nature Communications

Article Title: Immune checkpoint inhibitor-induced severe epidermal necrolysis mediated by macrophage-derived CXCL10 and abated by TNF blockade

doi: 10.1038/s41467-024-54180-7

Figure Lengend Snippet: a The secretion of CXCL10, TNF, GNLY, and GZMB were measured by ELISA in the blister fluids of 6 patients with ICI-SJS/TEN and 15 burn patients (Burns). b Plasma expression levels of CXCL10, CXCL9, TNF, IFN-γ, IL-6, IL-8, GNLY, and GZMB were measured by cytokine array or ELISA in 8 patients with ICI-induced SJS/TEN (ICI-SJS/TEN), 32 patients with ICI-induced mild cADR (ICI-mild cADR), and 9 control participants (Ctrl; including 4 participants with ICI-tolerant and 5 with HD). Plasma from patients with ICI-induced cADR were obtained during the acute stage of cADR phenotypes. Data in ( a ) and ( b ) are presented as the mean ± SD. P values were calculated by a two-sided unpaired Mann–Whitney U test. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: The following primary antibodies were used: rabbit monoclonal anti-human CXCL10-FITC antibody (1:100 dilution, LS-C123903, LSBio), mouse monoclonal anti-human CD163 antibody (1:100 dilution, ab156769, Abcam), mouse monoclonal anti-human CD8 antibody (1:100 dilution, ab199016, Abcam), and rabbit polyclonal anti-human CXCR3 antibody (1:200 dilution, ab71864, Abcam).

Techniques: Enzyme-linked Immunosorbent Assay, Expressing, Control, MANN-WHITNEY

Journal: Nature Communications

Article Title: Immune checkpoint inhibitor-induced severe epidermal necrolysis mediated by macrophage-derived CXCL10 and abated by TNF blockade

doi: 10.1038/s41467-024-54180-7

Figure Lengend Snippet: a scRNA-seq were performed for lesional BC and PBMC samples from 5 patients with ICI-induced SJS/TEN and 3 patients with drug-induced SJS/TEN. Ranking of the significant and relevant differentially expressed genes (DEG) in macrophage/monocyte/myeloid cells comparing between ICI-SJS/TEN lesional BC and drug-induced SJS/TEN lesional BC. b Ranking of the significant and relevant DEG in macrophage/monocyte/myeloid cells comparing between ICI-SJS/TEN PBMC and drug-SJS/TEN PBMC. c Ranking of the significant and relevant DEG in CD8 + T cells comparing between ICI-SJS/TEN lesional BC and drug-SJS/TEN lesional BC. d Ranking of the significant and relevant DEG in CD8 + T cells comparing between ICI-SJS/TEN PBMC and drug-SJS/TEN PBMC. e scRNA-seq analysis of PBMC collected during the recovery stage of a patient with ICI-induced SJS/TEN (ICI-SJS/TEN-1) after ex vivo culture with phosphate-buffered saline (PBS, solvent control), ICI stimulation (ICI), and ICI plus anti-TNF (ICI+anti-TNF). f Identification of cell clusters across sample groups. g Frequencies of different immune cells in each cluster across groups treated with PBS, ICI, and ICI+anti-TNF. The Violin plots display canonical marker genes for these immune cell clusters are shown in Supplementary Fig. . h Violin plots display the expression of selected ICI-induced SJS/TEN-associated genes, including GZMB , CD163 , TNF , CXCR3 , and CXCL10 , in CD8 + T cell or macrophage/monocyte cell clusters after ex vivo culture. i The macrophage/monocyte phenotype after ex vivo culture was measured by flow cytometric assay. The percentage of CD163 + CXCL10 + cells was determined in CD14 + monocyte cells (detailed gating information for flow cytometric assay is shown in Supplementary Fig. ). i - 1 The populations of CD163 + CXCL10 + cells in CD14 + monocytes after ex vivo treatment. j Cytotoxic CD8 + T cell phenotypes after ex vivo culture were measured. GranzymeB (GZMB) + cells were determined in CD8 + T cells (detailed gating information for the flow cytometric assay is shown in Supplementary Fig. ). j - 1 The populations of GZMB + cells in CD8 + T cells identified in PBMC after ex vivo treatment are shown. k Expression levels of CXCL10 in ex vivo cultured supernatant were detected. The significance of DEG was defined a using a two-sided non-parametric Wilcoxon rank-sum test and Bonferroni correction. Data in ( i – j ) are presented with boxplot showing individual data points, the first quartile, the median, and the third quartile. A black * indicates that P values were calculated by two-sided unpaired Mann–Whitney U test; a blue * indicates that P values were calculated by two-sided paired Wilcoxon matched-pairs signed-rank test.

Article Snippet: The following primary antibodies were used: rabbit monoclonal anti-human CXCL10-FITC antibody (1:100 dilution, LS-C123903, LSBio), mouse monoclonal anti-human CD163 antibody (1:100 dilution, ab156769, Abcam), mouse monoclonal anti-human CD8 antibody (1:100 dilution, ab199016, Abcam), and rabbit polyclonal anti-human CXCR3 antibody (1:200 dilution, ab71864, Abcam).

Techniques: Ex Vivo, Saline, Solvent, Control, Marker, Expressing, Flow Cytometry, Cell Culture, MANN-WHITNEY

Journal: Nature Communications

Article Title: Immune checkpoint inhibitor-induced severe epidermal necrolysis mediated by macrophage-derived CXCL10 and abated by TNF blockade

doi: 10.1038/s41467-024-54180-7

Figure Lengend Snippet: a 10X Genomic single-cell RNA sequencing (scRNA-seq) analysis in PBMC from three patients with ICI-induced SJS-TEN. UMAP plot generated from a merged dataset of the three patients with ICI-SJS/TEN pre- and post-TNF blockade treatment (total n = 31,904 cells). b UMAP plots segregated according to cellular origin. Dark red indicates the cell clusters before TNF blockade, whereas green indicates those after TNF blockade. Dotted regions highlight cytotoxic cells (including CD8 + T cells and NK/NKT cells) and myeloid/monocyte clusters both before and after TNF blockade. c Pie charts of three ICI-SJS/TEN patients showing relative cluster abundances before and after TNF blockade. The populations of total myeloid/monocyte cells before and after TNF blockade are shown in red. d , e Tumor images in the ICI-induced SJS-TEN overlapping case (ICI-SJS/TEN-1) with metaplastic breast carcinoma were evaluated by computed tomography (CT), revealing a durable complete response after recovery from the SJS/TEN episode. The red arrow indicates the tumor location. f Serial plasma CXCL10, TNF, and GZMB levels in 6 available patients with ICI-induced SJS/TEN during the clinical course from acute to late recovery stages. Day 1 on the horizontal axis is defined as the first day of hospitalization. The arrow in each line chart indicates TNF blockade (etanercept) administration. * of CXCL10 level represent P value calculated by two-sided paired Wilcoxon matched-pairs signed-rank test. g Clinical course, skin presentation, main tumor evaluation by CT, and serial plasma CXCL10 and TNF levels for one patient with ICI-induced SJS/TEN (ICI-SJS/TEN-15) who tolerated to continuous the same type of ICI immunotherapy combined with concurrent TNF blockade. Day 1 on the horizontal axis is defined as the first day of hospitalization. The blue arrow indicates TNF blockade (etanercept) administration time points, and the red arrow indicates the ICI (atezolizumab) treatment and re-challenge time points. The dotted line, arrow range, and values in the CT image indicate the tumor size.

Article Snippet: The following primary antibodies were used: rabbit monoclonal anti-human CXCL10-FITC antibody (1:100 dilution, LS-C123903, LSBio), mouse monoclonal anti-human CD163 antibody (1:100 dilution, ab156769, Abcam), mouse monoclonal anti-human CD8 antibody (1:100 dilution, ab199016, Abcam), and rabbit polyclonal anti-human CXCR3 antibody (1:200 dilution, ab71864, Abcam).

Techniques: RNA Sequencing Assay, Generated, Computed Tomography

Journal: Frontiers in Veterinary Science

Article Title: Advancing the early detection of canine cognitive dysfunction syndrome with machine learning-enhanced blood-based biomarkers

doi: 10.3389/fvets.2024.1390296

Figure Lengend Snippet: Protein profiling of the serum from APP/PS1 mice and MPTP-induced mice for disease model animals was obtained by performing proteome array analysis. (A) Images of array spots on the membrane are proteome array for immune blot, including 111 cytokine, chemokine, and growth factor types. The red rectangle indicates RBP4 and CXCL10 showing expression levels in the APP/PS1 and MPTP-induced mice. Magnifying indicates an enlarged image of RBP4 and CXCL10 in the square below each image. Array spots were analyzed according to the manufacturer’s instructions. (B) Quantifying RBP4 and CXCL10 levels in control and disease model animals from serum. Data are mean ± standard error of the mean (SEM). * , p < 0.05; ** , p < 0.01; *** , p < 0.001 vs. control by a Student’s two-tailed t-test. AD: Alzheimer’s disease; PD: Parkinson’s disease.

Article Snippet: The following antibodies were used: monoclonal mouse anti-RBP4 antibody (orb751184, Biorbyt, Berkeley, CA, United States); polyclonal rabbit anti-CXCL10 antibody (abx104024, Abbexa, Cambridge, United Kingdom); polyclonal rabbit anti-NOX4 antibody (NB110-58849, Novusbio, Centennial, CO, United States); polyclonal rabbit anti-transferrin antibody (NBP1-97472, Novusbio, Centennial, CO, United States); peroxidase labeled horse anti-mouse IgG (H + L) (7076P2, Cell Signaling Technology, Danvers, MA, United States); peroxidase labeled goat anti-rabbit IgG (H + L) (PI-1000, Vector Laboratories, Burlingame, CA, United States).

Techniques: Membrane, Expressing, Control, Two Tailed Test

Journal: Frontiers in Veterinary Science

Article Title: Advancing the early detection of canine cognitive dysfunction syndrome with machine learning-enhanced blood-based biomarkers

doi: 10.3389/fvets.2024.1390296

Figure Lengend Snippet: RBP4, CXCL10, and NOX4 protein levels in dogs of normal and CCDS groups. (A) Representative immunoblot analysis of RBP4, CXCL10, and NOX4 expression in CCDS groups compared to normal groups. (B) Quantifying RBP4, CXCL10, and NOX4 levels in normal and CCDS groups from plasma. Data are mean ± SEM. * , p < 0.05 vs. normal by a Student’s two-tailed t -test.

Article Snippet: The following antibodies were used: monoclonal mouse anti-RBP4 antibody (orb751184, Biorbyt, Berkeley, CA, United States); polyclonal rabbit anti-CXCL10 antibody (abx104024, Abbexa, Cambridge, United Kingdom); polyclonal rabbit anti-NOX4 antibody (NB110-58849, Novusbio, Centennial, CO, United States); polyclonal rabbit anti-transferrin antibody (NBP1-97472, Novusbio, Centennial, CO, United States); peroxidase labeled horse anti-mouse IgG (H + L) (7076P2, Cell Signaling Technology, Danvers, MA, United States); peroxidase labeled goat anti-rabbit IgG (H + L) (PI-1000, Vector Laboratories, Burlingame, CA, United States).

Techniques: Western Blot, Expressing, Clinical Proteomics, Two Tailed Test

Journal: Frontiers in Veterinary Science

Article Title: Advancing the early detection of canine cognitive dysfunction syndrome with machine learning-enhanced blood-based biomarkers

doi: 10.3389/fvets.2024.1390296

Figure Lengend Snippet: RBP4, CXCL10, and NOX4 levels were measured in the plasma from animal groups diagnosed with mild cognitive impairment (MCI) and severe cognitive impairment (SCI) based on CCDR scores. CCDS represents a group inclusive of both MCI and SCI, separated by a dotted line. (A) The bar graphs represent the quantification of RBP4, CXCL10, and NOX4 ELISA levels in each group. Data are mean ± SEM. (B) Representative violin plot graphs of the distribution of biomarker levels in each group. The dot in the graph revealed the distribution of individual samples, and the lines in the violin shape represent quartiles and medians. * , p < 0.05; ** , p < 0.01; *** , p < 0.001; **** , p < 0.0001 vs. each group by a one-way ANOVA. ### , p < 0.001; #### , p < 0.0001 vs. normal by a Student’s two-tailed t -test.

Article Snippet: The following antibodies were used: monoclonal mouse anti-RBP4 antibody (orb751184, Biorbyt, Berkeley, CA, United States); polyclonal rabbit anti-CXCL10 antibody (abx104024, Abbexa, Cambridge, United Kingdom); polyclonal rabbit anti-NOX4 antibody (NB110-58849, Novusbio, Centennial, CO, United States); polyclonal rabbit anti-transferrin antibody (NBP1-97472, Novusbio, Centennial, CO, United States); peroxidase labeled horse anti-mouse IgG (H + L) (7076P2, Cell Signaling Technology, Danvers, MA, United States); peroxidase labeled goat anti-rabbit IgG (H + L) (PI-1000, Vector Laboratories, Burlingame, CA, United States).

Techniques: Clinical Proteomics, Enzyme-linked Immunosorbent Assay, Biomarker Discovery, Two Tailed Test

Journal: Frontiers in Veterinary Science

Article Title: Advancing the early detection of canine cognitive dysfunction syndrome with machine learning-enhanced blood-based biomarkers

doi: 10.3389/fvets.2024.1390296

Figure Lengend Snippet: The correlation heatmap manifested the correlation between five variables: RBP4, CXCL10, NOX4, CCDR, and age. The color of each cell indicates a high correlation in red and a low correlation in blue, with the darker color indicating the strength of the correlation.

Article Snippet: The following antibodies were used: monoclonal mouse anti-RBP4 antibody (orb751184, Biorbyt, Berkeley, CA, United States); polyclonal rabbit anti-CXCL10 antibody (abx104024, Abbexa, Cambridge, United Kingdom); polyclonal rabbit anti-NOX4 antibody (NB110-58849, Novusbio, Centennial, CO, United States); polyclonal rabbit anti-transferrin antibody (NBP1-97472, Novusbio, Centennial, CO, United States); peroxidase labeled horse anti-mouse IgG (H + L) (7076P2, Cell Signaling Technology, Danvers, MA, United States); peroxidase labeled goat anti-rabbit IgG (H + L) (PI-1000, Vector Laboratories, Burlingame, CA, United States).

Techniques:

Journal: Frontiers in Veterinary Science

Article Title: Advancing the early detection of canine cognitive dysfunction syndrome with machine learning-enhanced blood-based biomarkers

doi: 10.3389/fvets.2024.1390296

Figure Lengend Snippet: Machine learning results summary for the normal and MCI.

Article Snippet: The following antibodies were used: monoclonal mouse anti-RBP4 antibody (orb751184, Biorbyt, Berkeley, CA, United States); polyclonal rabbit anti-CXCL10 antibody (abx104024, Abbexa, Cambridge, United Kingdom); polyclonal rabbit anti-NOX4 antibody (NB110-58849, Novusbio, Centennial, CO, United States); polyclonal rabbit anti-transferrin antibody (NBP1-97472, Novusbio, Centennial, CO, United States); peroxidase labeled horse anti-mouse IgG (H + L) (7076P2, Cell Signaling Technology, Danvers, MA, United States); peroxidase labeled goat anti-rabbit IgG (H + L) (PI-1000, Vector Laboratories, Burlingame, CA, United States).

Techniques:

Journal: Frontiers in Veterinary Science

Article Title: Advancing the early detection of canine cognitive dysfunction syndrome with machine learning-enhanced blood-based biomarkers

doi: 10.3389/fvets.2024.1390296

Figure Lengend Snippet: ROC curve graph between each group obtained through machine learning. (A) ROC curve for the combination of RBP4 and NOX4 between normal and MCI using the SVM algorithm. AUC is shown as 0.83. (B) ROC curve for the combination of RBP4, CXCL10, and NOX4 between normal and SCI using the Extra Tree algorithm. AUC is shown as 0.78. (C) ROC curve for the combination of RBP4 and NOX4 between normal and CCDS using the SVM algorithm. AUC is shown as 0.75.

Article Snippet: The following antibodies were used: monoclonal mouse anti-RBP4 antibody (orb751184, Biorbyt, Berkeley, CA, United States); polyclonal rabbit anti-CXCL10 antibody (abx104024, Abbexa, Cambridge, United Kingdom); polyclonal rabbit anti-NOX4 antibody (NB110-58849, Novusbio, Centennial, CO, United States); polyclonal rabbit anti-transferrin antibody (NBP1-97472, Novusbio, Centennial, CO, United States); peroxidase labeled horse anti-mouse IgG (H + L) (7076P2, Cell Signaling Technology, Danvers, MA, United States); peroxidase labeled goat anti-rabbit IgG (H + L) (PI-1000, Vector Laboratories, Burlingame, CA, United States).

Techniques:

Journal: Frontiers in Veterinary Science

Article Title: Advancing the early detection of canine cognitive dysfunction syndrome with machine learning-enhanced blood-based biomarkers

doi: 10.3389/fvets.2024.1390296

Figure Lengend Snippet: Machine learning results summary for the normal and SCI.

Article Snippet: The following antibodies were used: monoclonal mouse anti-RBP4 antibody (orb751184, Biorbyt, Berkeley, CA, United States); polyclonal rabbit anti-CXCL10 antibody (abx104024, Abbexa, Cambridge, United Kingdom); polyclonal rabbit anti-NOX4 antibody (NB110-58849, Novusbio, Centennial, CO, United States); polyclonal rabbit anti-transferrin antibody (NBP1-97472, Novusbio, Centennial, CO, United States); peroxidase labeled horse anti-mouse IgG (H + L) (7076P2, Cell Signaling Technology, Danvers, MA, United States); peroxidase labeled goat anti-rabbit IgG (H + L) (PI-1000, Vector Laboratories, Burlingame, CA, United States).

Techniques:

Journal: Frontiers in Veterinary Science

Article Title: Advancing the early detection of canine cognitive dysfunction syndrome with machine learning-enhanced blood-based biomarkers

doi: 10.3389/fvets.2024.1390296

Figure Lengend Snippet: Machine learning results summary for the normal and CCDS.

Article Snippet: The following antibodies were used: monoclonal mouse anti-RBP4 antibody (orb751184, Biorbyt, Berkeley, CA, United States); polyclonal rabbit anti-CXCL10 antibody (abx104024, Abbexa, Cambridge, United Kingdom); polyclonal rabbit anti-NOX4 antibody (NB110-58849, Novusbio, Centennial, CO, United States); polyclonal rabbit anti-transferrin antibody (NBP1-97472, Novusbio, Centennial, CO, United States); peroxidase labeled horse anti-mouse IgG (H + L) (7076P2, Cell Signaling Technology, Danvers, MA, United States); peroxidase labeled goat anti-rabbit IgG (H + L) (PI-1000, Vector Laboratories, Burlingame, CA, United States).

Techniques: