Journal: bioRxiv

Article Title: Identification of CD317-Positive Pro-inflammatory Immune Stromal Cells in Human Mesenchymal Stromal Cell Preparations

doi: 10.1101/2022.02.10.479972

Figure Lengend Snippet: (A) Comparative mRNA expression of ICAM-1 in MSC lines and primary cells sorted by CD317 expression (RNA was extracted from 3 different donors or 5 cell line passages; qPCR performed in triplicate, mean shown ± SEM). (B) Mean fluorescence intensity of ICAM-1 expression on the cell surface of MSC lines and primary MSCs differentially gated by CD317 staining (MSCs from 5 different donors or 4 different passages of MSC lines were stained for flow cytometry, mean shown ± SEM). (C)/(D) Comparative (mean ± SEM) mRNA expression of CXCL10 (red) and CXCL11 (blue) in MSC lines/ primary MSCs sorted for CD317 expression (RNA was extracted from 7 different donors/7 different cell passages; experiments were performed in triplicate). (E/F) CXCL10 secretion by MSC lines prior to IFN-γ priming and after priming with baseline (unprimed) secretion subtracted (mean ± SEM, n=2). (G/H) Comparative mRNA expression of 8 IFN-γ signature genes in MSC lines/primary MSCs sorted by CD317 expression (RNA was extracted from 5 different donors/5 different cell passages; experiments were performed in triplicate, mean shown ± SEM). (I)/(J) IFN-γ score for MSC lines/primary MSCs sorted by CD317 expression (n=5)*/** = significance at P<0.05/0.01 using an appropriate statistical test.

Article Snippet: To detect secreted proteins, supernatants from 100,000 cells incubated in 2.5 ml of serum free DMEM for 24 hours was analysed for secreted proteins by enzyme-linked immunosorbent assays (ELISA) using ELISA kits for CXCL10, CXCL11 (BioLegend); CCL2 (eBioscience); and SAA4 (Stratech) following manufacturers instructions.

Techniques: Expressing, Fluorescence, Staining, Flow Cytometry

Journal: Hepatology (Baltimore, Md.)

Article Title: Hepatitis C virus induces interferon-? and interferon-stimulated genes in primary liver cultures

doi: 10.1002/hep.24580

Figure Lengend Snippet: Genes induced following HCV infection in HFLC Results of 4 analyses on two independent livers are shown. Liver AB-080510 and AE-102910 were each infected with both Clone 2 and Jc1G. Values shown represent the peak fold increase in mRNA levels for each gene, relative to uninfected cultures from the same liver at the same time point. See Figure S2 for kinetics of gene induction. Expression of IFNα1, IFNω, IFNγ, IL1β, IL6, IL8, MDK, MOV10, DDIT4, c15orf8, and c8orf4 was not detected at any time point. In three of four comparisons, the relative gene induction in response to Clone 2 was dramatically higher than that in response to Jc1G, as shown for AB-080510. In one of four comparisons (AE-102910, shown here), high levels of gene induction were observed in response to both Jc1G and Clone 2.

Article Snippet: CXCL10 and CXCL11 ELISAs (R&D Systems, Minneapolis, MN) were performed according to the manufacturer’s instructions.

Techniques: Infection, Expressing, Transduction

Journal: Hepatology (Baltimore, Md.)

Article Title: Hepatitis C virus induces interferon-? and interferon-stimulated genes in primary liver cultures

doi: 10.1002/hep.24580

Figure Lengend Snippet: HFLC (AB-080510) were infected with HCVcc Clone 2. At the indicated times, supernatants were harvested for ELISA and cells were harvested for RNA preparation. HCV RNA is indicated in copies/culture (mean ± SD) on a log10 scale. IL29, CXCL10, and CXCL11 mRNA are shown as fold change (mean ± SD) over untreated cultures at the same time point, also on a log10 scale. Proteins are indicated in pg/ml (mean ± SD) of culture supernatant on a log10 scale. (A), HCV RNA. (B), mRNA and protein for IL29 (left), CXCL10 (middle) and CXCL11 (right); one experiment is shown out of five (IL29) or two (CXCL10 and CXCL11) with similar results.

Article Snippet: CXCL10 and CXCL11 ELISAs (R&D Systems, Minneapolis, MN) were performed according to the manufacturer’s instructions.

Techniques: Infection, Enzyme-linked Immunosorbent Assay

Journal: Hepatology (Baltimore, Md.)

Article Title: Hepatitis C virus induces interferon-? and interferon-stimulated genes in primary liver cultures

doi: 10.1002/hep.24580

Figure Lengend Snippet: DNA was prepared from the available material (organ identification numbers are indicated above each graph) and subjected to rs12979860 SNP genotyping as described in Materials and Methods. Cultures were infected with Jc1G or Clone 2, as indicated. Data shown represent HCV RNA levels and induction of IL29, CXCL10, and CXCL11 RNA. (A), Samples with CC genotype. Note that IL29 protein (135 ±100 pg/ml) was detected in supernatants from AB-082710 at 96 hours, but no 72-hour RNA sample was available for RT-PCR. (B), Samples with TT genotype. (C), Samples with CT genotype. Data shown represent 15 experiments using 11 organs.

Article Snippet: CXCL10 and CXCL11 ELISAs (R&D Systems, Minneapolis, MN) were performed according to the manufacturer’s instructions.

Techniques: Infection, Reverse Transcription Polymerase Chain Reaction

Journal: BioFactors (Oxford, England)

Article Title: Pancreatic β-cell Production of CXCR3 Ligands Precedes Diabetes Onset

doi: 10.1002/biof.1304

Figure Lengend Snippet: A. Cxcl9, Cxcl10 and Cxcl11 transcript levels were measured in islets isolated from 10 week old male (n = 4) and female NOD mice (n = 5). Data are expressed relative to age-matched BALB/c control mice (n = 7). **p<0.01, *p<0.05. B–F. Islets from Wistar rats (B, C; n = 4 per group) or human islets (D–F; n =3 per group) were untreated (NT) or stimulated with 10 ng/mL IL-1β, 100 U/mL IFN-γ or both cytokines for 3 h. B–F. Relative mRNA abundance of CXCL9 (B, D), CXCL10 (E) and CXCL11 (C, F) was determined by RT-PCR. ***p<0.001 vs. NT, **p<0.01 vs. NT, *p<0.05 vs. NT.

Article Snippet: Serum levels of CXCL9, CXCL10, and CXCL11 were detected using the mouse CXCL9 Quantikine ELISA kit (Cat # MCX900), CXCL10 Quantikine ELISA kit (Cat # MCX100) and the mouse CXCL11 DuoSet ELISA kit (Cat # DY572), all from R&D Systems (Minneapolis, MN) using the protocol provided by the manufacturer.

Techniques: Isolation, Control, Reverse Transcription Polymerase Chain Reaction

Journal: BioFactors (Oxford, England)

Article Title: Pancreatic β-cell Production of CXCR3 Ligands Precedes Diabetes Onset

doi: 10.1002/biof.1304

Figure Lengend Snippet: A–C. 832/13 rat insulinoma cells were stimulated with 1 ng/mL IL-1β or IL-1β plus 100 U/mL IFN-γ for the indicated times (NT; no treatment). D–F. 832/13 cells were pretreated for 1 h with either DMSO or 0.5 μg/mL Cycloheximide (CHX). Cells were subsequently exposed to IL-1β (1 ng/mL) or the combination of IL-1β and IFN-γ (100 U/mL) for 2 h. Cellular mRNA levels of Cxcl9 (A, D), Cxcl10 (B, E) and Cxcl11 (C, F) were detected by RT-PCR. n.s. = not significant vs respective treatment in DMSO control group. Data are shown as means ± SEM from three independent experiments.

Article Snippet: Serum levels of CXCL9, CXCL10, and CXCL11 were detected using the mouse CXCL9 Quantikine ELISA kit (Cat # MCX900), CXCL10 Quantikine ELISA kit (Cat # MCX100) and the mouse CXCL11 DuoSet ELISA kit (Cat # DY572), all from R&D Systems (Minneapolis, MN) using the protocol provided by the manufacturer.

Techniques: Reverse Transcription Polymerase Chain Reaction, Control

Journal: BioFactors (Oxford, England)

Article Title: Pancreatic β-cell Production of CXCR3 Ligands Precedes Diabetes Onset

doi: 10.1002/biof.1304

Figure Lengend Snippet: A. 832/13 cells were exposed to 100 U/mL IFN-γ for the indicated times. PO4-STAT1Y701 and total STAT1 protein abundance were determined by immunoblotting. B, C. 832/13 cells were pre-treated for 1 h with increasing concentrations of JAKi (1 nM, 10 nM, 100 nM), followed by a 3 h stimulation with IL-1β alone (1 ng/mL) or IL-1β plus 100 U/mL IFN-γ. ***p<0.001 vs. DMSO (black bar), *p<0.05 vs. DMSO (black bar). D, E. 832/13 cells were transfected with two siRNA duplexes targeting STAT1 using a scrambled siRNA sequence duplex as a control. 48 h post- transfection cells were cultured for 3 h with 1 ng/ml IL-1β or IL-1β plus 100 U/ml IFN-γ. ***p<0.001 vs. siScramble (black bar), *p<0.05 vs. siScramble (black bar). Cxcl9 (B, D) and Cxcl11 (C, E) mRNA levels were quantified. Data are represented as means ± SEM from three independent experiments. The immunoblot in A was repeated on two separate occasions.

Article Snippet: Serum levels of CXCL9, CXCL10, and CXCL11 were detected using the mouse CXCL9 Quantikine ELISA kit (Cat # MCX900), CXCL10 Quantikine ELISA kit (Cat # MCX100) and the mouse CXCL11 DuoSet ELISA kit (Cat # DY572), all from R&D Systems (Minneapolis, MN) using the protocol provided by the manufacturer.

Techniques: Quantitative Proteomics, Western Blot, Transfection, Sequencing, Control, Cell Culture

Journal: BioFactors (Oxford, England)

Article Title: Pancreatic β-cell Production of CXCR3 Ligands Precedes Diabetes Onset

doi: 10.1002/biof.1304

Figure Lengend Snippet: A. 832/13 cells were transduced with adenoviruses encoding either βGAL, wild-type STAT1 (WT), STAT1Y701F, STAT1S727A, STAT1Y701F/S727A (DM; double mutant) or STAT1S727T. STAT1 abundance was determined by immunoblotting. B, C. 832/13 cells were transduced with the adenoviruses indicated in (A); 24 h post-transduction cells were stimulated for 3 h with either IL-1β (1 ng/mL) alone or IL-1β plus IFN-γ (100 U/mL). *p<0.05, #p<0.1. D, E. Rat islets were transduced with the indicated adenoviruses. 24 h post-transduction cells were stimulated with both IL-1β (10 ng/mL) and IFN-γ (100 U/mL) for 3 h. ***p<0.001, **p<0.01. Relative mRNA abundance of Cxcl9 (B, D) and Cxcl11 (C, E) was determined by RT-PCR. Date are expressed as means ± SEM from 3 (B, C) or 2 (D, E) individual experiments. The immunoblot in A was repeated on two individual occasions.

Article Snippet: Serum levels of CXCL9, CXCL10, and CXCL11 were detected using the mouse CXCL9 Quantikine ELISA kit (Cat # MCX900), CXCL10 Quantikine ELISA kit (Cat # MCX100) and the mouse CXCL11 DuoSet ELISA kit (Cat # DY572), all from R&D Systems (Minneapolis, MN) using the protocol provided by the manufacturer.

Techniques: Transduction, Mutagenesis, Western Blot, Reverse Transcription Polymerase Chain Reaction

Journal: BioFactors (Oxford, England)

Article Title: Pancreatic β-cell Production of CXCR3 Ligands Precedes Diabetes Onset

doi: 10.1002/biof.1304

Figure Lengend Snippet: Schematic representation of distal and proximal GAS sites in the Cxcl9 and Cxcl11 promoters are shown (left panels). Arrows are the diagrammatic depiction of PCR amplicons. A–D. 832/13 cells were stimulated with 100 U/mL IFN-γ for either 20 mins (middle panels) or a time course (right panels). ChIP assays were performed to determine relative occupancy of total STAT1 (middle panels) and PO4-STAT1Y701 (right panels) on the Cxcl9 proximal (A) and distal promoter (B), and on the Cxcl11 proximal (C) and distal (D) promoter. ***p<0.001 vs. NT, **p<0.01 vs. NT, *p<0.05 vs. NT. Data are expressed as means ± SEM from 3–4 individual experiments.

Article Snippet: Serum levels of CXCL9, CXCL10, and CXCL11 were detected using the mouse CXCL9 Quantikine ELISA kit (Cat # MCX900), CXCL10 Quantikine ELISA kit (Cat # MCX100) and the mouse CXCL11 DuoSet ELISA kit (Cat # DY572), all from R&D Systems (Minneapolis, MN) using the protocol provided by the manufacturer.

Techniques:

Journal: Immunity, Inflammation and Disease

Article Title: Clinical significance of serum CXCL9, CXCL10, and CXCL11 in patients with lupus nephritis

doi: 10.1002/iid3.1368

Figure Lengend Snippet: Demographic and clinical factors for lupus nephritis (LN) among patients with systemic lupus erythematosus (SLE).

Article Snippet: The concentrations of serum CXCL9, CXCL10, and CXCL11 were detected using ELISA kits purchased from R&D Biosystems.

Techniques: Activity Assay

Journal: Immunity, Inflammation and Disease

Article Title: Clinical significance of serum CXCL9, CXCL10, and CXCL11 in patients with lupus nephritis

doi: 10.1002/iid3.1368

Figure Lengend Snippet: Comparisons of serum CXCL9 (A), CXCL10 (B), and CXCL11 (C) among systemic lupus erythematosus patients with lupus nephritis (SLE‐LN, n = 68) or not (SLE, n = 92) and healthy controls (HC, n = 70). Data were shown with mean ± SD. *** p < .001. Brown–Forsythe ANOVA test followed by Dunnett's T3 multiple comparisons test. CXCL, C‐X‐C motif chemokine ligand; SLE‐LN, systemic lupus erythematosus‐lupus nephritis.

Article Snippet: The concentrations of serum CXCL9, CXCL10, and CXCL11 were detected using ELISA kits purchased from R&D Biosystems.

Techniques:

Journal: Immunity, Inflammation and Disease

Article Title: Clinical significance of serum CXCL9, CXCL10, and CXCL11 in patients with lupus nephritis

doi: 10.1002/iid3.1368

Figure Lengend Snippet: Pearson correlation analysis of serum CXCL9 with CXCL10 (A), CXCL9 with CXCL11 (B), CXCL10 with CXCL11 (C) in all the enrolled systemic lupus erythematosus patients ( n = 160). p < .001 for all. CXCL, C‐X‐C motif chemokine ligand.

Article Snippet: The concentrations of serum CXCL9, CXCL10, and CXCL11 were detected using ELISA kits purchased from R&D Biosystems.

Techniques:

Journal: Immunity, Inflammation and Disease

Article Title: Clinical significance of serum CXCL9, CXCL10, and CXCL11 in patients with lupus nephritis

doi: 10.1002/iid3.1368

Figure Lengend Snippet: ROC analysis of serum CXCL9 (A), CXCL10 (B), CXCL11 (C), and their combined test model (D) for the diagnosis of lupus nephritis (LN) among patients with systemic lupus erythematosus (SLE). CXCL, C‐X‐C motif chemokine ligand; ROC, receiver operating characteristic.

Article Snippet: The concentrations of serum CXCL9, CXCL10, and CXCL11 were detected using ELISA kits purchased from R&D Biosystems.

Techniques:

Journal: Immunity, Inflammation and Disease

Article Title: Clinical significance of serum CXCL9, CXCL10, and CXCL11 in patients with lupus nephritis

doi: 10.1002/iid3.1368

Figure Lengend Snippet: According to the Austin activity index, there were 44 inactive LN patients and 24 active LN patients. Comparisons of serum CXCL9 (A), CXCL10 (B), and CXCL11 (C) between inactive LN patients and active LN patients. * p < .05, ** p < .01 from Unpaired t‐ test with Welch's correction. CXC, C‐X‐C motif chemokine ligand; LN, lupus nephritis.

Article Snippet: The concentrations of serum CXCL9, CXCL10, and CXCL11 were detected using ELISA kits purchased from R&D Biosystems.

Techniques: Activity Assay

Journal: PLoS ONE

Article Title: MMP-9 and CXCL8/IL-8 Are Potential Therapeutic Targets in Epidermolysis Bullosa Simplex

doi: 10.1371/journal.pone.0070123

Figure Lengend Snippet: A. SQRT-PCR shows increased matrix metalloproteinase mRNA expression in EBS-DM cell lines (n = 4). B. MMP-9 ELISA of 48-h-conditioned cell culture supernatant shows 2-fold upregulation of MMP-9 in KEB-7 and 46-fold upregulation in EBDM-1 at the protein level (n = 4). C. MMP-9 levels were highly increased in EBS patients blister fluids compared to healthy controls (n = 3 to n = 4). The numbers correlate with . Student`s t -test was performed with p values: * ≤0.05, ** ≤0.01, *** ≤0.005, ΔΔ≤0.0005, ΔΔΔ≤0.0001. (In 2C, the Student’s t -test compared the entire patient group to the entire control group).

Article Snippet: Protein levels of KLK5, MMP7, MMP9, CXCL1, CXCL8/IL-8, CXCL11 and CXCL14 were determined in 48-h-conditioned cell culture supernatant and in patients blister fluids by using Quantikine® ELISA (Human KLK5, # DKK500, R&D Systems; Human MMP7, # DMP700, R&D Systems; Human MMP9, # DMP900, R&D Systems; Human CXCL1/GROα, # DGR00, R&D Systems; Human CXCL8/IL-8, # D8000C, R&D Systems; Human CXCL11/I-TAC, # DCX110, Human CXCL14/BRAK, # DY866, R&D Systems) following the manufacturer’s protocol.

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Cell Culture, Control

Journal: PLoS ONE

Article Title: MMP-9 and CXCL8/IL-8 Are Potential Therapeutic Targets in Epidermolysis Bullosa Simplex

doi: 10.1371/journal.pone.0070123

Figure Lengend Snippet: A. SQRT-PCR of chemokine mRNA expression in NEB-1, KEB-7 and EBDM-1 (n = 3 to n = 5). CXCL1 , CXCL8/IL-8 and CXCL14 expression was increased in KEB-7. Only CXCL11 and CXCL14 were increased in EBDM-1. B. CXCL8/IL-8 ELISA of 48-h-conditioned cell culture supernatant showed 2-fold upregulation at the protein level in KEB-7 but not in EBDM-1, which correlates with the SQRT-PCR results (n = 4). C. CXCL8/IL-8 concentrations were highly increased in EBS patients blister fluids. In blister fluids of healthy controls no CXCL8/IL-8 was detectable (n = 3 to n = 4). The numbers correlate with . Student’s t -test was performed with p values: * ≤0.05, *** ≤0.005, Δ≤0.001, ‡ = no significant difference between investigated cell lines. (In 6C, the Student’s t -test compared the entire patient group to the entire control group).

Article Snippet: Protein levels of KLK5, MMP7, MMP9, CXCL1, CXCL8/IL-8, CXCL11 and CXCL14 were determined in 48-h-conditioned cell culture supernatant and in patients blister fluids by using Quantikine® ELISA (Human KLK5, # DKK500, R&D Systems; Human MMP7, # DMP700, R&D Systems; Human MMP9, # DMP900, R&D Systems; Human CXCL1/GROα, # DGR00, R&D Systems; Human CXCL8/IL-8, # D8000C, R&D Systems; Human CXCL11/I-TAC, # DCX110, Human CXCL14/BRAK, # DY866, R&D Systems) following the manufacturer’s protocol.

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Cell Culture, Control

Journal: Indian Journal of Dermatology

Article Title: The Effect of Transplantation of Cultured Autologous Melanocytes on CXCL9, CXCL10 and CXCL11 Expressions in Vitiligo

doi: 10.4103/ijd.ijd_925_22

Figure Lengend Snippet: Expression levels of serum CXCL9, CXCL10 and CXCL11 in different types of vitiligo patients

Article Snippet: Before the intervention, 5 ml of peripheral venous blood (PBMC) was collected from all the participants to detect the concentrations of CXCL9, CXCL10 and CXCL11 by ELISA kit (Wuhan, Boster Biological Technology, China).

Techniques: Expressing

Journal: Indian Journal of Dermatology

Article Title: The Effect of Transplantation of Cultured Autologous Melanocytes on CXCL9, CXCL10 and CXCL11 Expressions in Vitiligo

doi: 10.4103/ijd.ijd_925_22

Figure Lengend Snippet: Expression levels of serum CXCL9, CXCL10 and CXCL11 in vitiligo patients and healthy controls

Article Snippet: Before the intervention, 5 ml of peripheral venous blood (PBMC) was collected from all the participants to detect the concentrations of CXCL9, CXCL10 and CXCL11 by ELISA kit (Wuhan, Boster Biological Technology, China).

Techniques: Expressing

Journal: Indian Journal of Dermatology

Article Title: The Effect of Transplantation of Cultured Autologous Melanocytes on CXCL9, CXCL10 and CXCL11 Expressions in Vitiligo

doi: 10.4103/ijd.ijd_925_22

Figure Lengend Snippet: Expressions levels of serum CXCL9, CXCL10 and CXCL11 in vitiligo patients of different stages

Article Snippet: Before the intervention, 5 ml of peripheral venous blood (PBMC) was collected from all the participants to detect the concentrations of CXCL9, CXCL10 and CXCL11 by ELISA kit (Wuhan, Boster Biological Technology, China).

Techniques:

Journal: Indian Journal of Dermatology

Article Title: The Effect of Transplantation of Cultured Autologous Melanocytes on CXCL9, CXCL10 and CXCL11 Expressions in Vitiligo

doi: 10.4103/ijd.ijd_925_22

Figure Lengend Snippet: Expression levels of serum CXCL9, CXCL10 and CXCL11 in stable vitiligo before and after the operation

Article Snippet: Before the intervention, 5 ml of peripheral venous blood (PBMC) was collected from all the participants to detect the concentrations of CXCL9, CXCL10 and CXCL11 by ELISA kit (Wuhan, Boster Biological Technology, China).

Techniques: Expressing