mouse anti human cxcl10  (R&D Systems)

Journal: Science Advances

Article Title: Targeting HMGB2 acts as dual immunomodulator by bolstering CD8 + T cell function and inhibiting tumor growth in hepatocellular carcinoma

doi: 10.1126/sciadv.ads8597

Figure Lengend Snippet: ( A ) Representative images of harvested Hepa1-6 subcutaneous HCC tumors. Scale bar, 1 cm ( B ) Tumor growth curves of Hepa1-6 subcutaneous tumors in BALB/C nude mice and C57BL/6J mice. ( C ) Tumor weights of subcutaneous Hepa1-6 tumors in BALB/C nude mice and C57BL/6J mice ( n = 6). ( D ) Difference of tumor volumes between shCtrl and shHmgb2 subcutaneous tumors in BALB/C nude mice and C57BL/6J mice ( n = 6). ( E ) Differential GO pathways in Hepa1-6 shHmgb2 cells. JAK, Janus kinase. ( F ) GSEA analysis shows top pathway enriched in Hepa1-6 shHmgb2 cells. ( G ) Western blotting experiment shows STAT1 pathway changes in Hepa1-6 cells and Huh7 cells. Cells were treated with IFN-γ (10 ng/ml) or fludarabine (10 μM) for 24 hours. ( H ) Annexin V apoptosis analysis for Hepa1-6 cells treated with vehicle and IFN-γ (10 or 20 ng/ml). PI, propidium iodide. ( I ) Quantification for proportions of apoptotic cells after treatment of IFN-γ. ( J ) T cell killing assay with Hepa1-6 shCtrl and shHmgb2 cells and wild-type (WT) CD8 + T cells ( n = 5). RLU, relative light unit. ( K ) Terminal deoxynucleotidyl transferase–mediated deoxyuridine triphosphate nick end labeling (TUNEL) staining and quantification of Hepa1-6 subcutaneous tumors ( n = 4). Scale bar, 25 μm. HPF, high power field. ( L ) Representative immunohistochemistry images of CD8, IFN-γ, and CXCL10 staining in Hepa1-6 subcutaneous tumors. Scale bars, 20 μm. ( M ) Quantification of CD8, IFN-γ, and CXCL10 staining in Hepa1-6 subcutaneous tumors ( n = 4). Data are presented as the means ± SEM. * P < 0.05; ** P < 0.01; *** P < 0.001. Two-way ANOVA test for (B) Student’s t test for (C), (D), (I), (J), (K), and (M).

Article Snippet: Anti-human/mouse CXCL10 , 10937-1-AP , Proteintech, China.

Techniques: Western Blot, End Labeling, TUNEL Assay, Staining, Immunohistochemistry

Journal: Science Advances

Article Title: Targeting HMGB2 acts as dual immunomodulator by bolstering CD8 + T cell function and inhibiting tumor growth in hepatocellular carcinoma

doi: 10.1126/sciadv.ads8597

Figure Lengend Snippet: Markers and article numbers of antibodies. PE, phycoerythrin; FITC, fluorescein isothiocyanate; APC, antigen-presenting cell; HRP, horseradish peroxidase; mAb, monoclonal antibody.

Article Snippet: Anti-human/mouse CXCL10 , 10937-1-AP , Proteintech, China.

Techniques: Ubiquitin Proteomics, Purification, In Vivo

Journal: Science Advances

Article Title: Targeting HMGB2 acts as dual immunomodulator by bolstering CD8 + T cell function and inhibiting tumor growth in hepatocellular carcinoma

doi: 10.1126/sciadv.ads8597

Figure Lengend Snippet: Names and sequence of primers. F, forward; R, reverse.

Article Snippet: Anti-human/mouse CXCL10 , 10937-1-AP , Proteintech, China.

Techniques: Sequencing

Journal: medRxiv

Article Title: Characterization of CXCL10 as a biomarker of respiratory tract infections detectable by open-source lateral flow immunoassay

doi: 10.1101/2024.01.12.24301261

Figure Lengend Snippet: A: Schematic of workflow. Pre-processed transcriptomic datasets using the Harmonizome resource (Rouillard et al ., 2006) database of processed microarray datasets under the category “GEO Signatures of Differentially Expressed Genes for Viral Infections” (Edgar et al ., 2002; Barrett et al ., 2013) was accessed for hypothesis testing for elevations in CXCL10 , CXCL11 , and TNFSF10 following viral exposure in vitro from a variety of experimental conditions. This dataset contained 366 individual datasets of mRNA expression profiles using microarray technology. All non-human experiments and non-respiratory viruses were excluded which filtered down to 199 microarray datasets. The studies in which CXCL10, CXCL11 and TNFSF10 appeared to be differentially expressed were counted (*Found in the top 300 or bottom 300 differentially expressed genes with a Harmonizome standard value greater than 1 (up-regulated) or below -1 (down-regulated).) Datasets included but not limited to Calu-3 cell lines, HAE cultures infected with respiratory viruses including but not limited to FluA, SARS-CoV and Human metapneumovirus. B: CXCL10 was found to be upregulated in 39 independent viral infection datasets, and downregulated in one. CXCL11 was upregulated in 33 viral infection datasets. TNFSF10 was upregulated in 36 and downregulated in 3. C: The housekeeping genes GAPDH , TUBB and ACTB were analyzed in all 199 and showed different expression levels in fewer datasets than for CXCL10 , CXCL11 , and TNFSF10 . GADPH was found to be upregulated in 1 independent viral infection datasets, and downregulated in 6. TUBB was down in 3 viral infection datasets. ACTB was upregulated in 5 and downregulated in 9.

Article Snippet: A recombinant anti-human CXCL10 mouse IgG-monoclonal (MAB2661), anti-mouse IgG goat-polyclonal (AF-266-NA), and recombinant CXCL0 protein (Product 266-IP) were selected from R&D Systems (Toronto, Ontario, Canada).

Techniques: Microarray, In Vitro, Expressing, Infection

Journal: medRxiv

Article Title: Characterization of CXCL10 as a biomarker of respiratory tract infections detectable by open-source lateral flow immunoassay

doi: 10.1101/2024.01.12.24301261

Figure Lengend Snippet: A: Schematic of workflow. B: From Yu et al ., 2019 - GSE117827: CXCL10 , CXCL11 and TNFSF10 gene expression was compared from the mid-turbinate nasal swab of pediatric subjects with RSV, RV and negative controls. Clustered heatmap of log 2 expression levels annotated by symptomatic, sex and infection with blue representing decreased expression and red increased expression. On the right, boxplot of RMA normalized expression (log 2 ) (n = 24). There was no significant difference when comparing rhinovirus infected NPS to healthy control gene expression for CXCL10 , CXCL11 and TNFSF10 (p > 0.05). CXCL10 and CXCL11 up-regulation was positively correlated with Respiratory Syncytial Virus (RSV) when compared to healthy control (p = 0.016, p = 0.006). C: Hamilton Regional Laboratory Medicine Program (HRLMP) microarray data: CXCL10 , CXCL11 and TNFSF10 gene expression was compared from the NPS of subjects with influenza A (FluA) and negative controls. Clustered heatmap of log 2 expression levels annotated by sex and infection status with blue representing decreased expression and red increased expression. On the right, boxplot of RMA normalized expression (log 2 ) (n = 22). No significant difference between CXCL10 , CXCL11 and TNFSF10 expression in FluA infection compared to negative control (p > 0.05). D: From Lieberman et al ., 2020 - GSE152075: CXCL10 , CXCL11 and TNFSF10 gene expression was compared from the NPS of individuals with suspected SARS-CoV-2 infection. Clustered heatmap of log 2 expression levels annotated by sex and infection status with blue representing decreased expression and red increased expression. On the right, boxplot of RMA normalized expression (log 2 ) (n = 484). CXCL10 , CXCL11 and TNFSF10 expression was significantly upregulated SARS-Cov-2 infection compared to those who tested negative (p < 0.001). E: From Mick et al ., 2020 - GSE156063: CXCL10 , CXCL11 and TNFSF10 gene expression was compared from the NPS of subjects SARS-CoV-2 positive, SARS-CoV-2 negative but positive for another respiratory virus and no respiratory virus detected by metagenomic next generation sequencing (i.e., non-viral ARI such as bacterial infection). Clustered heatmap of log 2 expression levels annotated by sex and infection status with blue representing decreased expression and red increased expression. On the right, boxplot of RMA normalized expression (log 2 ) (n = 234). CXCL10 , CXCL11 and TNFSF10 expression was significantly upregulated SARS-Cov-2 infection compared to healthy control (p < 0.001). * = p < 0.05, ** p < 0.01 and *** = p < 0.001.

Article Snippet: A recombinant anti-human CXCL10 mouse IgG-monoclonal (MAB2661), anti-mouse IgG goat-polyclonal (AF-266-NA), and recombinant CXCL0 protein (Product 266-IP) were selected from R&D Systems (Toronto, Ontario, Canada).

Techniques: Gene Expression, Expressing, Infection, Control, Virus, Microarray, Negative Control, Next-Generation Sequencing

Journal: medRxiv

Article Title: Characterization of CXCL10 as a biomarker of respiratory tract infections detectable by open-source lateral flow immunoassay

doi: 10.1101/2024.01.12.24301261

Figure Lengend Snippet: A: Schematic of workflow. B : SARS-CoV-2 viral sequencing reads and qPCR cycle thresholds correlate with the CXCL10/CXCL11/TNFSF10 gene signature. (N = 735). The “Viral Level Continuous” comparison group converted qRT-PCR cycle threshold (Ct) values into a continuous variable by inverting CT values where Ct = 15 is equal to 1.0 and a Ct > 40 is 0, CXCL10 showed an upregulation of log 2 fold change of 6.2 (q value = 1.23E-54), CXCL11 showed an upregulation of log 2 fold change of 6.0 (q value = 5.17 E-47) and TNFSF10 showed an upregulation of log 2 fold change of 1.9 (q value = 4.26E-38). Data and Figure from Butler et al . 2021 - For research purposes only. All rights reserved. © Mason Lab and Weill Cornell Medicine, 2020). C: Mortality of COVID-19 patients is associated with only modest changes in the CXCL10/CXCL11/TNFSF10 gene signature at the time of original patient sampling. No significant correlation was found (p > 0.05).

Article Snippet: A recombinant anti-human CXCL10 mouse IgG-monoclonal (MAB2661), anti-mouse IgG goat-polyclonal (AF-266-NA), and recombinant CXCL0 protein (Product 266-IP) were selected from R&D Systems (Toronto, Ontario, Canada).

Techniques: Sequencing, Comparison, Quantitative RT-PCR, Sampling

Journal: medRxiv

Article Title: Characterization of CXCL10 as a biomarker of respiratory tract infections detectable by open-source lateral flow immunoassay

doi: 10.1101/2024.01.12.24301261

Figure Lengend Snippet: A: Schematic of workflow. B: CXCL10 , CXCL11 , and TNFSF10 expression over time in hospitalized COVID-19 positive patients expressed as % change from first sampling (set as time=0). The data was collected from 6 independent patients (COVXXX) who had distinct sampling counts dependent on clinical management of COVID-19 infection. COV005 = 5 measurements, COV006 = 3 measurements, COV007 = 5 measurements, COV010 = 6 measurements, COV011 = 10 measurements, COV013 = 11 measurements. C: To determine which gene fluctuated the least of the course sampling, the variance of RMA values for CXCL10/CXCL11/TNFSF10 were calculated for each patient (colours) and averaged (grey). CXCL11 variance = 0.17, CXCL10 variance = 0.21 and TNFSF10 variance = 1.68. * p < 0.05 relative to mean variance for CXCL10 and CXCL11 .

Article Snippet: A recombinant anti-human CXCL10 mouse IgG-monoclonal (MAB2661), anti-mouse IgG goat-polyclonal (AF-266-NA), and recombinant CXCL0 protein (Product 266-IP) were selected from R&D Systems (Toronto, Ontario, Canada).

Techniques: Expressing, Sampling, Infection

Journal: medRxiv

Article Title: Characterization of CXCL10 as a biomarker of respiratory tract infections detectable by open-source lateral flow immunoassay

doi: 10.1101/2024.01.12.24301261

Figure Lengend Snippet: A: Schematic of workflow. B-C: From Grassl et al ., 2016 ultra deep analysis of the healthy saliva proteome, the CXCL10 protein was not found amongst the list of 5562 identified proteins (Supplementary Table 2). D: CXCL10 Levels in saliva of SARS-CoV-2 in hospitalized COVID-19 patients quantified using the Human Cytokine Array / Chemokine Array 71-plex (Eve Technologies, Calgary, Alberta, Canada). Healthy volunteers without symptoms of a respiratory infection were used as the control group. Mean concentration of CXCL10 in saliva was 86.4 pg/mL (SD = 109.6, n = 6) in healthy subjects, while a mean of 1186.6 pg/mL (SD = 1252.3) was observed in COVID-19 patients. The COVID-19 group showed a significantly greater CXCL10 concentration (* = p < 0.05).

Article Snippet: A recombinant anti-human CXCL10 mouse IgG-monoclonal (MAB2661), anti-mouse IgG goat-polyclonal (AF-266-NA), and recombinant CXCL0 protein (Product 266-IP) were selected from R&D Systems (Toronto, Ontario, Canada).

Techniques: Infection, Control, Concentration Assay

Journal: medRxiv

Article Title: Characterization of CXCL10 as a biomarker of respiratory tract infections detectable by open-source lateral flow immunoassay

doi: 10.1101/2024.01.12.24301261

Figure Lengend Snippet: A: Schematic of workflow. B-C: Sensitivity development and validation in ideal buffer (10mM HEPES, 150 mM NaCl, 0.1% Tween-20, 1% BSA, and 0.5% PEG 8000 – see methods for more details). Limit of detection of CXCL10 using a hand-held reader. A positive signal is generated at 2ng/mL with n = 5. D-E: Sensitivity test in artificial saliva (product #1700-0316, ASTM E2721-16 with Mucin, pH 7.0; Pickering Laboratories, Mountain View, California, USA) prepared with equal parts lateral flow buffer and artificial saliva - see methods for more details. F: Real world testing in human saliva from healthy control without (neat) and with (spiked) CXCL10 (10ng/mL) addition.

Article Snippet: A recombinant anti-human CXCL10 mouse IgG-monoclonal (MAB2661), anti-mouse IgG goat-polyclonal (AF-266-NA), and recombinant CXCL0 protein (Product 266-IP) were selected from R&D Systems (Toronto, Ontario, Canada).

Techniques: Biomarker Discovery, Generated, Control

Journal: Cells

Article Title: Induction of CXCL10-Mediated Cell Migration by Different Types of Galectins

doi: 10.3390/cells10020274

Figure Lengend Snippet: Galectins induce CXCL10 in a cell type-specific manner. ( A ) SDS-polyacrylamide gel electrophoresis of recombinant human galectins (rhGal) stained with Coomassie Blue. Molecular weights of protein standards are indicated on the left. The hemagglutination assay was performed using serial two-fold dilutions of the recombinant protein. The bottom row contains protein incubated with β-lactose, a competitive carbohydrate inhibitor of galectin binding. ( B ) Cultures of human corneal fibroblasts, human corneal epithelial cells, THP-1 monocytes and human umbilical vein endothelial cells (HUVECs) were incubated with 50 µg/mL rhGal-1, -3 and -8 or 50 µg/mL IFN-γ. Quantitative real-time PCR was used to determine the levels of CXCL10 mRNA after 6 h of incubation ( n = 3 independent experiments), whereas ELISA was used to determine the levels of CXCL10 protein in cell culture supernatants after 24 h of incubation ( n = 6 independent experiments). The data represent the mean ± SEM.

Article Snippet: A mouse monoclonal antibody against CXCL10 (1.2 μg/mL; MAB266, R&D Systems) was added to the lower compartment of the chamber in neutralization studies.

Techniques: Polyacrylamide Gel Electrophoresis, Recombinant, Staining, Hemagglutination Assay, Incubation, Binding Assay, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Cell Culture

Journal: Cells

Article Title: Induction of CXCL10-Mediated Cell Migration by Different Types of Galectins

doi: 10.3390/cells10020274

Figure Lengend Snippet: Galectins differentially dictate fibroblast-dependent immune cell chemotaxis. ( A ) Recombinant human galectin (rhGal)-1, -3 and -8 were incubated with fibroblasts for 24 h. The conditioned media were pre-cleared with β-lactose Sepharose beads three times to remove galectins prior to chemotaxis assays. The presence of galectins in pre-cleared conditioned media was evaluated by immunoblotting. ( B ) Quantification by flow cytometry of the migration of CFSE-labeled THP-1 and Jurkat cells to conditioned media from fibroblasts treated for 24 h with increasing concentrations of recombinant galectins or BSA ( n = 3 independent experiments performed in duplicate). ( C ) Migration of CFSE-labeled THP-1 and Jurkat cells to conditioned media from fibroblasts treated for 24 h with 50 μg/mL of recombinant galectins or BSA. A CXCL10 neutralizing antibody was added to the conditioned media in neutralization studies ( n = 4–5 independent experiments in duplicate). The data in ( B ) represent the mean ± SEM. The box and whisker plots show the 25 and 75 percentiles (box), the median, and the minimum and maximum data values (whiskers). Significance was determined using Student’s t test. ** p < 0.01.

Article Snippet: A mouse monoclonal antibody against CXCL10 (1.2 μg/mL; MAB266, R&D Systems) was added to the lower compartment of the chamber in neutralization studies.

Techniques: Chemotaxis Assay, Recombinant, Incubation, Western Blot, Flow Cytometry, Migration, Labeling, Neutralization, Whisker Assay

Journal: Journal of Virology

Article Title: Human Nasal Turbinate Tissues in Organ Culture as a Model for Human Cytomegalovirus Infection at the Mucosal Entry Site

doi: 10.1128/JVI.01258-20

Figure Lengend Snippet: Nasal mucosal tissue transcriptional response to HCMV infection. RNA was extracted from mock- and HCMV-infected tissues at 24 hpi and subjected to transcriptome analysis. To account for the potential donors’ tissue-to-tissue variability, two pools (each representing a mixture of 5 independent donor tissues) for each experimental condition were used, together representing 10 tissues from different individuals. (A) MA plot (log ratio versus abundance) comparing gene expression profiles in mock- and HCMV-infected tissues for all 26,340 queried genes. Genes of higher abundance in infected tissues are indicated by positive changes, and those of lower abundance in infected tissues are indicated by negative changes. Red dots denote significant genes (adjusted P [Padj] < 0.1); orange dots denote added nonsignificant genes (see Materials and Methods). (B) Heat map representation of all differentially expressed and added genes (red and orange dots in panel A), comparing the two pools of mock- and HCMV-infected tissues. Normalized expression values were scaled at gene level (scale is shown at top-right), then hierarchically clustered and drawn as a heat map. Representative upregulated innate immunity genes further analyzed by qRT-PCR as shown in panel F and downregulated epithelial-cell related genes are indicated. (C) Selected antiviral and proinflammatory innate immunity genes with a strong (>3-fold) upregulation in HCMV-infected versus mock-infected tissues. (D) AQP5 and MUC2 mRNA expression levels in HCMV infected tissues relative to the normalized levels (normalized to a value of 1) in mock-infected tissues. (E and F) The indicated viral/cellular mRNA levels were analyzed by qRT-PCR and normalized by cellular β-actin. The fold change between HCMV-infected and mock-infected tissues (F) is shown for each cellular gene in 6 independent nasal turbinate tissues (T1 to T6) obtained from different individuals. (G and H) Functional analysis of conditioned medium (CM) recovered at 1 (E) and 5 (F) dpi from mock- and HCMV-infected nasal turbinate tissues (prepared as described in Materials and Methods). (G) Human foreskin fibroblasts (HFF) and human retinal pigmented epithelial cells (ARPE) cultures were pretreated overnight with conditioned medium (CM) and infected with HCMV. HCMV IE1 mRNA levels were analyzed by qRT-PCR at 24 hpi and normalized by cellular β-actin. (H) Peripheral blood leukocytes (PBL) transwell migration toward CM from mock- or HCMV-infected nasal turbinate cultures (treated with ganciclovir when indicated). CM samples were preincubated with no IgG (control) or with α-CXCL10 antibodies, as indicated. The number of migrated cells was determined by flow cytometry. The data shown are representative of at least three independent experiments. Significant changes are indicated as *, P < 0.05; **, P < 0.01; ***, P < 0.001. NT, nontreated; n.s., nonsignificant.

Article Snippet: In pretreatment experiments, when indicated, CM was incubated with 0.2 μg/ml mouse anti-human CXCL10 monoclonal antibodies (MAB266; R&D Systems) for 1 h at 37°C and 5% CO 2 , before the transwell migration assay.

Techniques: Infection, Gene Expression, Expressing, Quantitative RT-PCR, Functional Assay, Migration, Control, Flow Cytometry

Journal: Journal of Virology

Article Title: Human Nasal Turbinate Tissues in Organ Culture as a Model for Human Cytomegalovirus Infection at the Mucosal Entry Site

doi: 10.1128/JVI.01258-20

Figure Lengend Snippet: Primers and probes for real-time PCR analysis

Article Snippet: In pretreatment experiments, when indicated, CM was incubated with 0.2 μg/ml mouse anti-human CXCL10 monoclonal antibodies (MAB266; R&D Systems) for 1 h at 37°C and 5% CO 2 , before the transwell migration assay.

Techniques: Real-time Polymerase Chain Reaction, Sequencing, SYBR Green Assay