Journal: Clinical and Vaccine Immunology

Article Title: Increased CXC Ligand 10 Levels and Gene Expression in Type 1 Leprosy Reactions

doi: 10.1128/cvi.00042-11

Figure Lengend Snippet: FIG. 1. Circulating CXCL10 in leprosy patients and healthy con- trols. Each point represents one sample from a different individual. The groups were healthy controls, borderline tuberculoid without re- action (BT), borderline lepromatous without reaction (BL), polar lep- romatous without reaction (LL), BT with reaction (BT plus RR), BL with reaction (BL plus RR), and LL patients with T2R (erythema nodosum leprosum). For patients with reaction (BT plus RR, BL plus RR, and T2R), the sample was taken at the time of reaction, prior to treatment. For LL, n 6; for all other groups, n 10. The horizontal lines indicate median values. The median level of CXCL10 in LL patients with type 2 reaction was not significantly elevated (P 0.9).

Article Snippet: Tissue sections were stained by an indirect immunoperoxidase method using goat anti-human CXCL10 (R&D Systems, Inc.; 1:40) or rabbit anti-human CXCR3 (Sigma) (1:75) for 60 min, followed by biotinylated donkey anti-goat IgG or goat anti-rabbit IgG (both 1:40; Jackson ImmunoResearch Laboratories, Inc.) for 30 min. Peroxidase-conjugated avidin-biotin complexes (Vectastain Elite ABC Reagent kit) were applied for 30 min. All incubations were followed by three washes with phosphate-buffered saline isotonic buffer, pH 7.4.

Techniques:

Journal: Clinical and Vaccine Immunology

Article Title: Increased CXC Ligand 10 Levels and Gene Expression in Type 1 Leprosy Reactions

doi: 10.1128/cvi.00042-11

Figure Lengend Snippet: FIG. 2. Circulating CXCL10 in repeated samples from leprosy patients who developed T1R. Each graph depicts the measurements made from serum samples obtained at 1 month intervals in borderline tuberculoid (A, no. 1 to 10) and borderline lepromatous (B, no. 11 to 20) patients. All patients with clinical T1R received standard multidrug treatment starting at the first visit. The arrows indicate the visits at which T1R was diagnosed. The number above each time point indicates the dose of prednisone started on that day, tapered monthly as indicated; in some cases, corticosteroids were not used initially but were started 1 to 2 months after the initial clinical diagnosis of T1R. Each serum specimen was obtained before corticosteroid treatment was initiated. CXCL10 was significantly elevated during T1R in the 10 BT patients (A) (P 0.0006) and the 10 BL patients (B) (P 0.0001). In patients 12, 16, and 17 (B), T1R was diagnosed by histopathological criteria only; when they were excluded from the analysis, the association of T1R with CXCL10 remained significant (P 0.05).

Article Snippet: Tissue sections were stained by an indirect immunoperoxidase method using goat anti-human CXCL10 (R&D Systems, Inc.; 1:40) or rabbit anti-human CXCR3 (Sigma) (1:75) for 60 min, followed by biotinylated donkey anti-goat IgG or goat anti-rabbit IgG (both 1:40; Jackson ImmunoResearch Laboratories, Inc.) for 30 min. Peroxidase-conjugated avidin-biotin complexes (Vectastain Elite ABC Reagent kit) were applied for 30 min. All incubations were followed by three washes with phosphate-buffered saline isotonic buffer, pH 7.4.

Techniques: Biomarker Discovery

Journal: Clinical and Vaccine Immunology

Article Title: Increased CXC Ligand 10 Levels and Gene Expression in Type 1 Leprosy Reactions

doi: 10.1128/cvi.00042-11

Figure Lengend Snippet: FIG. 3. Expression levels of CXCL10 and IFN- genes in skin biopsy specimens from leprosy patients. (A to H) Real-time quantitative RT-PCR was performed on cDNA obtained from sequential skin biopsy specimens from individual leprosy patients who were placed on MDT, none of whom had T1R at the time of the first biopsy. Five patients (A to E) had clinical symptoms of T1R at the time of the second biopsy; two of these (C and D) had a third biopsy after the reaction had resolved. Three patients (F to H) had no clinical symptoms of T1R at the time of the initial or second biopsy. Data were obtained using the standard-curve method and were normalized using 18S rRNA values, repeated three times for all determinations. The results are presented as the mean standard deviation.

Article Snippet: Tissue sections were stained by an indirect immunoperoxidase method using goat anti-human CXCL10 (R&D Systems, Inc.; 1:40) or rabbit anti-human CXCR3 (Sigma) (1:75) for 60 min, followed by biotinylated donkey anti-goat IgG or goat anti-rabbit IgG (both 1:40; Jackson ImmunoResearch Laboratories, Inc.) for 30 min. Peroxidase-conjugated avidin-biotin complexes (Vectastain Elite ABC Reagent kit) were applied for 30 min. All incubations were followed by three washes with phosphate-buffered saline isotonic buffer, pH 7.4.

Techniques: Expressing, Quantitative RT-PCR, Standard Deviation

Journal: Journal of Neuro-Oncology

Article Title: Rebound growth of BRAF mutant pediatric glioma cells after MAPKi withdrawal is associated with MAPK reactivation and secretion of microglia-recruiting cytokines

doi: 10.1007/s11060-024-04672-9

Figure Lengend Snippet: Cytokine expression and increased AKT activity are independent mechanisms without a tumor cell intrinsic effect on rebound growth. ( a ) Luminex-based multiplex assay results showing cytokine secretion during withdrawal after treatment for five days with either DMSO (solvent control) or 5 nM dabrafenib. Data is shown as mean ± SD ( n = 3 independent biological replicates). Two-tailed unpaired t-test, *p-value ≤ 0.05 **p-value ≤ 0.01; no indication: not significant. ( b ) Kinase phosphorylation array of samples treated for five days with 5 nM dabrafenib or DMSO (solvent control) followed by 24 h withdrawal (wd). Arrays consist of two membranes, each target is detected in technical duplicates. Images shown are representatives of two biological replicates. ( c - d ) Western blot analysis of AKT phosphorylation after five days treatment with 5 nM dabrafenib (dabra) followed by up to 72 h withdrawal. Blots shown are representative of three independent biological replicates ( c ). Quantification ( d ) is relative to solvent control (DMSO; dashed line) and shown as mean ± SD ( n = 3 independent biological replicates). One-sample t-test, * p-value ≤ 0.05; no indication: not significant. ( e - f ) Western blot analysis of AKT activity markers after treatment with recombinant cytokines. Treatment for five days with DMSO or 5 nM dabrafenib (dabra) served as negative and positive control for western blots ( e ). Blots shown are representative of three biological replicates ( e ). Quantification ( f ) was done relative to untreated samples (dashed line) and is shown as mean ± SD ( n = 3 independent biological replicates). One-sample t-test; no indication: not significant. ( g ) RT-qPCR analysis of chemokine gene expression after five days treatment with 5 nM dabrafenib (dabra) alone or in combination with varying concentrations of alpelisib (alp). Quantification was done relative to dabrafenib only treatment (0; dashed line) and is shown as mean ± SD ( n = 3 independent biological replicates). One-sample t-test, *p-value ≤ 0.05 **p-value ≤ 0.01; no indication: not significant. ( h ) Viable cell counts during treatment with 5nM dabrafenib (dabra) alone or in combination with a combination of antibodies neutralizing CCL2 (0.5 µg/mL), CX3CL1 (0.25 µg/mL), CXCL10 (0.25 µg/mL) and CCL7 (0.1 ng/mL) (neuABs), IgG control (1 µg/mL), 5 µM alpelisib (alp) or 1 µM ipatasertib (ipa) followed by ten days of withdrawal. Dashed line indicates withdrawal timepoint. Viable cell counts are normalized to treatment start (-5d). Data is shown on a logarithmic scale (base 10) as mean ± SD of at least three biological replicates. ( i ) Viable cell counts during treatment with 5 nM dabrafenib (dabra) followed by withdrawal. During withdrawal cells were either untreated (= solvent) or treated for five days with a combination of antibodies neutralizing CCL2 (0.5 µg/mL), CX3CL1 (0.25 µg/mL), CXCL10 (0.25 µg/mL) and CCL7 (0.1 ng/mL) (neuABs), IgG control (1 µg/mL), 5 µM alpelisib (alp) or 1 µM ipatasertib (ipa), followed by five days of withdrawal. Dashed lines indicate withdrawal timepoints. Viable cell counts are normalized to treatment start (-5d). Data is shown on a logarithmic scale (base 10) as mean ± SD of at least three biological replicates

Article Snippet: For cell count experiments and western blot analysis, cells were treated with antibodies neutralizing CCL2 (0.5 μg/ml; cat. no. MAB279, R&D systems), CX3CL1 (0.25 μg/ml; cat. no. MAB3652, R&D systems), CXCL10 (0.25 μg/mL; cat. no. MAB266, R&D systems) and CCL7 (0.1ng/ml; cat. no. MAB282, R&D systems) or mouse IgG (cat. no. MAB002, R&D systems).

Techniques: Expressing, Activity Assay, Luminex, Multiplex Assay, Solvent, Control, Two Tailed Test, Phospho-proteomics, Western Blot, Recombinant, Positive Control, Quantitative RT-PCR, Gene Expression

Journal: Journal of Neuro-Oncology

Article Title: Rebound growth of BRAF mutant pediatric glioma cells after MAPKi withdrawal is associated with MAPK reactivation and secretion of microglia-recruiting cytokines

doi: 10.1007/s11060-024-04672-9

Figure Lengend Snippet: Increased cytokine secretion upon MAPKi treatment and withdrawal induces increased microglia attraction. ( a ) Schematic of transwell migration assay setup to investigate migration of HMC3 cells towards conditioned media (CM) collected from BT-40. ( b ) Transwell migration assay of HMC3 cells towards conditioned media (CM) collected from BT-40 cells after 24 h of withdrawal after five days treatment with DMSO (solvent control), 5nM dabrafenib (d) or 2.7 nM dabrafenib and 0.3 nM trametinib (d + t). 2% FCS serves as baseline control as CM contains 2% FCS, 0% FCS as negative control and 10% FCS as positive control. Quantification is shown as mean ± SD ( n = 3 independent biological replicates; 2 technical duplicates per condition; 10–12 randomly distributed images were quantified per transwell) relative to 2% FCS. One-sample t-test, * p-value ≤ 0.05; no indication: not significant. ( c ) Transwell migration assay of HMC3 cells towards CM collected 24 h after dabrafenib withdrawal (dabra wd) containing either antibodies neutralizing CCL2 (1 µg/mL), CX3CL1 (0.5 µg/mL), CXCL10 (0.5 µg/mL) and CCL7 (0.2 ng/mL) (neuABs) or IgG (2 µg/mL). 2% FCS serves as baseline control as CM contains 2% FCS, 0% FCS as negative control and 10% FCS as positive control. Quantification is shown as mean ± SD ( n = 3 independent biological replicates; 2 technical duplicates per condition; 10–12 randomly distributed images were quantified per transwell) relative to 2% FCS. One-sample t-test and two-tailed unpaired t-test (comparing IgG to neuABs), *p-value ≤ 0.05 ***p-value ≤ 0.001; no indication: not significant. ( d ) Representative fluorescence images showing HMC3 migrated through the transwell, nuclei were stained with DAPI. Scale bar = 50 µM. ( e - h ) RT-qPCR analysis of CX3CL1 ( e ), CXCL10 ( f ), CCL2 ( g ) and CCL7 ( h ) expression in BT-40 xenograft tumors after six days treatment with dabrafenib (dabra; 100 mg/kg, six doses, once daily) followed by three days of withdrawal (withdrawal). Samples from mice showing tumor progression (#4, #6; Fig. d) during dabrafenib treatment were excluded from the analysis. Quantification was done relative to the median of untreated samples (control). CX3CL1 was undetected in one sample (#2), CCL2 was undetected in two samples (#2, #7) and CCL7 was undetected in six samples (#2, #3, #5, #7, #8, #11), for these samples Ct values were set to 40 (max. number of cycles). Boxplots depict the median, first and third quartiles. Whiskers extend from the hinge to the largest/smallest value no further than 1.5 * IQR from the hinge (where IQR is the interquartile range). Two-tailed unpaired t-test; no indication: not significant (control: n = 3 mice, dabra: n = 4 mice, withdrawal: n = 6 mice)

Article Snippet: For cell count experiments and western blot analysis, cells were treated with antibodies neutralizing CCL2 (0.5 μg/ml; cat. no. MAB279, R&D systems), CX3CL1 (0.25 μg/ml; cat. no. MAB3652, R&D systems), CXCL10 (0.25 μg/mL; cat. no. MAB266, R&D systems) and CCL7 (0.1ng/ml; cat. no. MAB282, R&D systems) or mouse IgG (cat. no. MAB002, R&D systems).

Techniques: Transwell Migration Assay, Migration, Solvent, Control, Negative Control, Positive Control, Two Tailed Test, Fluorescence, Staining, Quantitative RT-PCR, Expressing

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: 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