Journal: PloS one

Article Title: Association of interleukin-6 signalling with the muscle stem cell response following muscle-lengthening contractions in humans.

doi: 10.1371/journal.pone.0006027

Figure Lengend Snippet: Figure 1. Circulating IL-6 and muscle IL-6 mRNA and IL-6 receptor response muscle lengthening contractions (MLC). (1a) Average serum interleukin-6 (IL-6) concentration. Time 0 hr corresponds to pre-intervention values; all other time-points correspond to post-intervention time (hr). (1b) Relative IL-6 mRNA expression, expressed as fold-change from 0 hr. (1c) Pearson correlation of serum IL-6 concentration versus muscle IL-6 mRNA (fold-change), correlation is representative of the individual data points and presented as mean values ($)6SD (error bars). (1d) Relative IL-6 receptor (IL-6Ra) mRNA expression, expressed as fold-change from 0 hr. (1e) Immunofluorescent image (406) of a muscle cross-section triple-stained for Pax7 (red), IL-6Ra (green) and nuclei (DAPI = blue); IL-6Ra staining is apparent on the sarcolemma and satellite cell membrane. White arrow denotes one of the Pax7+ nuclei which co-localized with IL-6Ra (scale bar = 100 mm). Values are reported as mean6S.E.M. Mean values represent the mean for all 8 subjects per time-point (8 samples per time-point, 40 samples total). *p,0.05 vs. 0 hr. doi:10.1371/journal.pone.0006027.g001

Article Snippet: Immunofluorescence 7 mm sections were cryosectioned and stained with antibodies against Pax7 (neat; DSHB, USA); IL-6 (500 ng/mL, MAB 2061, R&D Systems, USA); p-STAT3 (p-STAT3 Y705 1:100, Cell Signaling Technologies Inc., USA); IL-6Ra (1:50, MCA822, Serotec, UK); PCNA (1:200, ab15497, Abcam Inc., USA); and Laminin (1:1000, L8271, Sigma-Aldrich, Canada).

Techniques: Concentration Assay, Expressing, Staining, Membrane

Journal: medRxiv

Article Title: In severe COVID-19, SARS-CoV-2 induces a chronic, TGF-β-dominated adaptive immune response

doi: 10.1101/2020.09.04.20188169

Figure Lengend Snippet: Peripheral blood activated B cells (CD27 high CD38 high ) were isolated and sorted by FACS for single cell sequencing. a) Percentage of CD27 high CD38 high cells among live B cells. Each sample is represented by a circle. The line indicates the median. Unpaired two sided Mann-Whitney U test. b) UMAP representation of 72277 CD27 high CD38 high sorted B cells from 9 COVID-19 ICU patients and 3 healthy controls. Between 2416 and 11229 cells were recovered per sample. Transcriptionally similar clusters were identified using shared nearest neighbor (SNN) modularity optimization (left). The combination of CD19, MS4A1, CD27, CD38, IFIT1, MIKI67, IRF4 and PRDM1 expression were used for annotation of the different activation/differentiation stages (right). c) Heatmap of genes that resemble markers for the six different clusters. Depicted are genes with a p-value < 0.01 (Wilcoxon rank sum test) after bonferroni correction, and an average absolute fold-change > log2(1.3). Shown are z-scores of the average expression. d) UMAP representation of the expression levels of selected signature genes for activated/differentiated B cells. e) UMAP representation of analyzed cells from one healthy control and two ICU patients, representing early (first week, patient #1) and late (>7days, patient #5) phase after ICU admission. f) Percentage of cells belonging to a defined cluster among all sequenced B cells per donor. Each dot represents one time point from one single donor. Donors were grouped as “HC” for healthy controls (n = 3), “1st week” for patients within 7 days after ICU admission (n = 6) and “Late” for patients who have been admitted to the ICU for more than a week at the time of analysis (n = 8). The line indicates the median. Significance was determined by using a two-sided analysis of varience (ANOVA) folowed by Tukey’s multiple comparison test with corrected p values as indicated in the figure.

Article Snippet: Fluorescence-coupled antibodies used: IgA2-PE (Clone REA995, Miltenyi, Cat. No.130-117-763, dilution 1:50); CD27-PE (Clone REA499, Miltenyi, Cat. No. 130-114-166, dilution 1:50); CD38-PE (Clone IB6, Miltenyi, Cat. No. 130-113-427, dilution 1:50) and IgA-PE (Clone IS11–8E10, Miltenyi, Cat. No. 130-114-002, dilution 1:50).

Techniques: Isolation, Sequencing, MANN-WHITNEY, Expressing, Activation Assay, Control, Comparison

Journal: medRxiv

Article Title: In severe COVID-19, SARS-CoV-2 induces a chronic, TGF-β-dominated adaptive immune response

doi: 10.1101/2020.09.04.20188169

Figure Lengend Snippet: a) 5-marker MELC panel of SARS-CoV2-positive and control lungs (SARS-CoV2-negative). Respective patient characteristics are given in supplementary Fig. 4a. Each image of each patient depicts the same field of view of the same section, sequentially stained with the fluorescence-labelled antibodies indicated and the nuclear stain DAPI. Magenta arrows indicate IgA2+IgA+CD27 + CD38 + cells (containing a nucleus). Images contain 2048 × 2048 pixels and are generated using an inverted wide-field fluorescence microscope with a 20x objective, a lateral resolution of 325 nm and an axial resolution above 5 µm. Scale bar: 100 µm. b) Absolute numbers of IgA2+IgA+CD27 + CD38 + cells per field of view in all MELC runs acquired (two runs per patient except for COVID-19_A with four runs and Control_B with one single run; see and supplementary Fig. 4b). Each field of view is represented by a circle. The line indicates the median. Unpaired two sided Mann-Whitney U test. c) Region of interest of one exemplary control and COVID-19 lung (as in a)), showing an overlay of the indicated markers. White arrows point out IgA2+CD27 + CD38 + cells. Scale bar: 20 µm. (d-e) Bronchoalveolar lavage (BAL) cells for single cell sequencing were enriched for CD45 + cells via MACS and live cells were further sorted using FACS (see supplementary Fig. 4c). d) UMAP of 5459 cells representing clusters containing T cells, B cells and CD14 -expressing cells from patient #1 on day 59 following ICU admission. B and T cell cluster identification based on BCR/TCR, CD19, CD3E, CD4 and CD8A expression (see supplementary Fig. 4d). UMAP representation of expression of TGFB1, IL21, CD40LG and IFNG . e) UMAP coordinates and clustering was computed for 433 and 2723 cells from patient #9 on days 31 and 46 following ICU admission, respectively. UMAP representation of expression of TGFB1, IL21, CD40LG and IFNG at the two different time points is shown side by side.

Article Snippet: Fluorescence-coupled antibodies used: IgA2-PE (Clone REA995, Miltenyi, Cat. No.130-117-763, dilution 1:50); CD27-PE (Clone REA499, Miltenyi, Cat. No. 130-114-166, dilution 1:50); CD38-PE (Clone IB6, Miltenyi, Cat. No. 130-113-427, dilution 1:50) and IgA-PE (Clone IS11–8E10, Miltenyi, Cat. No. 130-114-002, dilution 1:50).

Techniques: Marker, Control, Staining, Fluorescence, Generated, Microscopy, MANN-WHITNEY, Sequencing, Expressing

Journal: Cell reports

Article Title: High-dimensional profiling clusters asthma severity by lymphoid and non-lymphoid status

doi: 10.1016/j.celrep.2021.108974

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Anti-Human/Mouse IL-5 (TRFK5)-143Nd , Fluidigm , Cat# 3143003B.

Techniques: Purification, Control, Antibody Labeling, Flow Cytometry, Software, Staining, Blocking Assay

Journal: mAbs

Article Title: Characterization of concurrent target suppression by JNJ-61178104, a bispecific antibody against human tumor necrosis factor and interleukin-17A

doi: 10.1080/19420862.2020.1770018

Figure Lengend Snippet: Summary of JNJ-8104, golimumab and CNTO 6785 in vitro target-binding kinetics and activities.

Article Snippet: The concentrations of total IL-17A in cyno and human serum were measured with similar MSD assays, except using a non-competing biotinylated mouse anti-human IL-17A antibody (Clone 4H1524.1, Rockland) for capture and a non-competing ruthenium-labeled mouse IgG1 anti-human IL-17A antibody (Clone 41802, R&D Systems) for detection.

Techniques: In Vitro

Journal: mAbs

Article Title: Characterization of concurrent target suppression by JNJ-61178104, a bispecific antibody against human tumor necrosis factor and interleukin-17A

doi: 10.1080/19420862.2020.1770018

Figure Lengend Snippet: Inhibition of in vitro inflammatory responses mediated by endogenous TNF and IL-17A. Human rheumatoid arthritis fibroblast-like synovial cells (RA-FLS) were cocultured with in vitro differentiated T h 17/T h 1 cells. Various concentrations of JNJ-8104, CNTO 9809 (parental anti-TNF), CNTO 4782 (parental anti-IL-17A) or equal-molar fixed-ratio combination of CNTO 9809 and CNTO 4782 were incubated with the coculture for 48 hours. Endogenous TNF and IL-17A-mediated production of IL-6 (a) and GROα (b) in the supernatants were measured.

Article Snippet: The concentrations of total IL-17A in cyno and human serum were measured with similar MSD assays, except using a non-competing biotinylated mouse anti-human IL-17A antibody (Clone 4H1524.1, Rockland) for capture and a non-competing ruthenium-labeled mouse IgG1 anti-human IL-17A antibody (Clone 41802, R&D Systems) for detection.

Techniques: Inhibition, In Vitro, Incubation

Journal: mAbs

Article Title: Characterization of concurrent target suppression by JNJ-61178104, a bispecific antibody against human tumor necrosis factor and interleukin-17A

doi: 10.1080/19420862.2020.1770018

Figure Lengend Snippet: Inhibition of rhTNF- and rhIL-17A-induced Cellular Influx by JNJ-8104 in the Airway Lumen of Mice. Mice were intranasally instilled with rhTNF- and rhIL-17A in combination. After 6 h their lungs were lavaged and total numbers of BAL cells (a) and neutrophils (b) were enumerated as detailed in “Materials & Methods.” Mice were injected intraperitoneally with the test mAbs or isotype control18 hours prior to cytokines challenge. Data are represented as mean ± SEM; N = 6–7 mice/group.

Article Snippet: The concentrations of total IL-17A in cyno and human serum were measured with similar MSD assays, except using a non-competing biotinylated mouse anti-human IL-17A antibody (Clone 4H1524.1, Rockland) for capture and a non-competing ruthenium-labeled mouse IgG1 anti-human IL-17A antibody (Clone 41802, R&D Systems) for detection.

Techniques: Inhibition, Injection

Journal: mAbs

Article Title: Characterization of concurrent target suppression by JNJ-61178104, a bispecific antibody against human tumor necrosis factor and interleukin-17A

doi: 10.1080/19420862.2020.1770018

Figure Lengend Snippet: Observed serum concentrations versus time profiles of Total mAb (a), total TNF (b), and Total IL-17A (c) following IV administration of golimumab (parental anti-TNF), CNTO 6785 (anti-IL-17A with identical parental Fab arms) or JNJ-8104 at the specified dose levels in cynomolgus monkeys. Data are represented as mean ± SD.

Article Snippet: The concentrations of total IL-17A in cyno and human serum were measured with similar MSD assays, except using a non-competing biotinylated mouse anti-human IL-17A antibody (Clone 4H1524.1, Rockland) for capture and a non-competing ruthenium-labeled mouse IgG1 anti-human IL-17A antibody (Clone 41802, R&D Systems) for detection.

Techniques:

Journal: mAbs

Article Title: Characterization of concurrent target suppression by JNJ-61178104, a bispecific antibody against human tumor necrosis factor and interleukin-17A

doi: 10.1080/19420862.2020.1770018

Figure Lengend Snippet: Estimated JNJ-8104, CNTO148 and CNTO 6785 cynomolgus monkey PK/TE model parameters (mean, residual standard error%) and predicted human parameters.

Article Snippet: The concentrations of total IL-17A in cyno and human serum were measured with similar MSD assays, except using a non-competing biotinylated mouse anti-human IL-17A antibody (Clone 4H1524.1, Rockland) for capture and a non-competing ruthenium-labeled mouse IgG1 anti-human IL-17A antibody (Clone 41802, R&D Systems) for detection.

Techniques:

Journal: mAbs

Article Title: Characterization of concurrent target suppression by JNJ-61178104, a bispecific antibody against human tumor necrosis factor and interleukin-17A

doi: 10.1080/19420862.2020.1770018

Figure Lengend Snippet: PK/TE model fitting of PK, total IL-17A and Free IL-17A profiles following IV administration of JNJ-8104 (a) and CNTO 6785 (b) in cynomolgus monkeys. Symbols = Observed individual data; Lines = Mean model prediction.

Article Snippet: The concentrations of total IL-17A in cyno and human serum were measured with similar MSD assays, except using a non-competing biotinylated mouse anti-human IL-17A antibody (Clone 4H1524.1, Rockland) for capture and a non-competing ruthenium-labeled mouse IgG1 anti-human IL-17A antibody (Clone 41802, R&D Systems) for detection.

Techniques:

Journal: mAbs

Article Title: Characterization of concurrent target suppression by JNJ-61178104, a bispecific antibody against human tumor necrosis factor and interleukin-17A

doi: 10.1080/19420862.2020.1770018

Figure Lengend Snippet: Comparison of observed and model-predicted Total IL-17A profiles following a single IV administration of JNJ-8104 at 0.1, 0.3, 1, 3 and 10 mg/kg (a−e) normal human subjects. Symbols = Observed individual data; Solid lines = Mean model prediction based on cyno PK/TE model parameters and allometric scaling for human PK; Dotted lines = Mean model prediction based on estimated JNJ-8104 human PK parameters from FIH data with cyno-based in vivo KD.

Article Snippet: The concentrations of total IL-17A in cyno and human serum were measured with similar MSD assays, except using a non-competing biotinylated mouse anti-human IL-17A antibody (Clone 4H1524.1, Rockland) for capture and a non-competing ruthenium-labeled mouse IgG1 anti-human IL-17A antibody (Clone 41802, R&D Systems) for detection.

Techniques: Comparison, In Vivo

Journal: Stem Cell Research & Therapy

Article Title: Bone marrow mesenchymal stem cells alleviate liver fibrosis after rat liver transplantation through JAK1/STAT5 pathway

doi: 10.1186/s13287-025-04353-y

Figure Lengend Snippet: BMSCs may alleviate liver fibrosis after liver transplantation through the JAK1/STAT5 pathway. ( A ) Box plot for FPKM values. PCA analysis. Bar plot of the statistic of differentially expressed genes. Venn graph of the differentially expressed genes. Volcano plot of the differentially expressed genes. Heat map of RNA differential expression. GO analysis reveals the potential impact of the differentially expressed RNAs on functions. KEGG analysis reveals the potential impact of the differentially expressed RNA on pathways. KEGG map of the JAK/STAT pathway. ( B ) Western blot analyses of JAK1, p-STAT5, STAT5, p-STAT1, STAT1, p-STAT3, STAT3, IL7R, and IL7 in liver tissue. TUBA1A was used as the loading control except for p-STAT5, p-STAT1, and p-STAT3. STAT5, STAT1, and STAT3 were used as the loading control for p-STAT5, p-STAT1, and p-STAT3, respectively. Jak1 , Il7r , and Il7 mRNA levels in liver tissue. ( C ) Microscopic images of each group stained with HE, Masson, Sirius Red, and ACTA2 (magnification: ×100, n = 6). Area stained with Masson, Sirius Red, and ACTA2, as well as Metavir fibrosis staging. The levels of serum ALT, AST, and TBil in each group. ( D ) Western blot analyses of COL1A1, JAK1, p-STAT5, STAT5, ACTA2, and CASP3 in liver tissues. TUBA1A was used as the loading control except for p-STAT5. STAT5 was used as the loading control for p-STAT5. Full-length blots/gels are presented in Supplementary Fig. . The samples were derived from the same experiment and processed in parallel. Col1a1 , Jak1 , Acta2 , and Casp3 mRNA levels in liver tissues. ACTA2, actin alpha 2; ALT, alanine aminotransferase; AST, aspartate aminotransferase; BMSCs, bone mesenchymal stem cells; CASP3, caspase 3; COL1A1, collagen type I alpha 1 chain; GO, Gene Ontology; IL, interleukin; JAK, Janus-activated kinase; KEGG, Kyoto Encyclopedia of Genes and Genomes; LT, liver transplantation; STAT, signal transducer and activator of transcription; TBil, total bilirubin. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001

Article Snippet: The LT + anti-IL7 group received anti-IL7 antibody (M25; Cat# HY-P990210, MCE, Shanghai, China) (250 μg/kg of body weight, per ip, once a week) for 2 weeks after LT.

Techniques: Transplantation Assay, Quantitative Proteomics, Western Blot, Control, Staining, Derivative Assay

Journal: Stem Cell Research & Therapy

Article Title: Bone marrow mesenchymal stem cells alleviate liver fibrosis after rat liver transplantation through JAK1/STAT5 pathway

doi: 10.1186/s13287-025-04353-y

Figure Lengend Snippet: BMSCs may inhibit the activation of HSC-T6 cells after hypoxia–reoxygenation and reduce their apoptosis through the JAK1/STAT5 pathway. ( A ) Western blot analyses of JAK1, p-STAT5, STAT5, p-STAT1, STAT1, p-STAT3, STAT3, IL7R, and IL7 in HSC-T6 cells. TUBA1A was used as the loading control except for p-STAT5, p-STAT1, and p-STAT3. STAT5, STAT1, and STAT3 were used as the loading control for p-STAT5, p-STAT1, and p-STAT3, respectively. Jak1 and Il7r mRNA levels in HSC-T6 cells. ( B ) Western blot analyses of COL1A1, JAK1, p-STAT5, STAT5, and ACTA2 in HSC-T6 cells. TUBA1A was used as the loading control except for p-STAT5. STAT5 was used as the loading control for p-STAT5. Col1a1 , Jak1 and Acta2 mRNA levels in HSC- T6 cells. ( C ) Apoptosis results of flow cytometry in each group. Full-length blots/gels are presented in Supplementary Fig. . The samples were derived from the same experiment and processed in parallel. ACTA2, actin alpha 2; BMSCs, bone mesenchymal stem cells; COL1A1, collagen type I alpha 1 chain; IL, interleukin; JAK, Janus-activated kinase; STAT, signal transducer and activator of transcription. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001

Article Snippet: The LT + anti-IL7 group received anti-IL7 antibody (M25; Cat# HY-P990210, MCE, Shanghai, China) (250 μg/kg of body weight, per ip, once a week) for 2 weeks after LT.

Techniques: Activation Assay, Western Blot, Control, Flow Cytometry, Derivative Assay

Journal: Stem Cell Research & Therapy

Article Title: Bone marrow mesenchymal stem cells alleviate liver fibrosis after rat liver transplantation through JAK1/STAT5 pathway

doi: 10.1186/s13287-025-04353-y

Figure Lengend Snippet: BMSCs regulate the IL7R/JAK1/STAT5 pathway in HSCs by modulating the secretion of IL7 by hepatic cells. ( A ) Microscopic images of each group stained with HE, Masson, Sirius Red, ACTA2, p-STAT5 and STAT5 (magnification: ×100, n = 6). Area stained with Masson, Sirius Red, ACTA2, and p-STAT5/STAT5, as well as Metavir fibrosis staging. The levels of serum ALT, AST, and TBil in each group. ( B ) Western blot analyses of COL1A1, JAK1, p-STAT5, STAT5 and ACTA2 in liver tissue. TUBA1A was used as the loading control except for p-STAT5. STAT5 was used as the loading control for p-STAT5. Col1a1 , Jak1 , and Acta2 mRNA levels in liver tissue. ( C ) Western blot analyses of IL7 in IAR20 cells. TUBA1A was used as the loading control. Il7 mRNA levels in IAR20 cells. ( D ) Western blot analyses of COL1A1, JAK1, p-STAT5, STAT5 and ACTA2 in HSC-T6 cells. TUBA1A was used as the loading control except for p-STAT5. STAT5 was used as the loading control for p-STAT5. Col1a1 , Jak1 and Acta2 mRNA levels in HSC- T6 cells. Full-length blots/gels are presented in Supplementary Fig. . The samples were derived from the same experiment and processed in parallel. ACTA2, actin alpha 2; ALT, alanine aminotransferase; AST, aspartate aminotransferase; BMSCs, bone mesenchymal stem cells; COL1A1, collagen type I alpha 1 chain; IL, interleukin; JAK, Janus-activated kinase; LT, liver transplantation; STAT, signal transducer and activator of transcription; TBil, total bilirubin. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001

Article Snippet: The LT + anti-IL7 group received anti-IL7 antibody (M25; Cat# HY-P990210, MCE, Shanghai, China) (250 μg/kg of body weight, per ip, once a week) for 2 weeks after LT.

Techniques: Staining, Western Blot, Control, Derivative Assay, Transplantation Assay