Journal: Genes

Article Title: Differentially Expressed Genes of Natural Killer Cells Can Distinguish Rheumatoid Arthritis Patients from Healthy Controls

doi: 10.3390/genes11050492

Figure Lengend Snippet: ELISA Quantification of ( A ) CXCL16 and ( B ) IL-1β and ( C ) IFN-γ in the plasma of RA patients and healthy controls.

Article Snippet: The kits used were human CXCL16 (DY1164), IFN-γ (DY285) and IL-1β (DY201) duosets (R&D systems, Minneapolis, MN, USA).

Techniques: Enzyme-linked Immunosorbent Assay, Clinical Proteomics

Journal: International immunopharmacology

Article Title: MiR-625-5p is a potential therapeutic target in sepsis by regulating CXCL16/CXCR6 axis and endothelial barrier.

doi: 10.1016/j.intimp.2024.112508

Figure Lengend Snippet: Fig. 1. Predictive value of biomarkers and clinical variables on cumulative ICU survival in patients with sepsis or septic shock patients. Blood samples were collected from patients withsepsis or septic shock and healthy subjects (control) for qRT-PCR analysis of miR-625-5p. miR-625-5p was normalized with the level of h-SNORD44 (internal control) to determine the ratios, and the ratios of the control were arbitrarily set at 1. The relative levels of miR-625-5p (a) and CXCL16 (b) were detected by qRT-PCR. The levels of CXCL16 (c), SDC-1 (d), HS (e), and VE-cadherin (f) were detected by using ELISA. The red rulers in Figures a and d represent mean values with SDs, quantitative data was compared with one-way ANOVA between three groups. The blue rulers in Figures b, c, e, and f represent the medians with ranges, quantitative data was compared with Kruskal–Wallis analysis between three groups. ROC curve analysis of APACHE II score, SOFA score, miR-625-5p, CXCL16, SDC-1, PCT, and lactate at admission for the prediction of 28-day mortality (g). Kaplan–Meier survival estimates for all patients with sepsis or septic shock according to the respective level of miR-625-5p (miR-625-5p, cut-of: 25) (h). Correlations between biomarker levels and various clinical parameters (i–n). *P < 0.05, **P < 0.01.

Article Snippet: Serum levels of syndecan-1 (EK1339, BOSTER, China), heparan sulfate (356350, Usbiological, USA), and VE-cadherin (DCADV0, R&D Systems, USA), and CXCL16 (EK0741, BOSTER, China) levels in the serum and cell supernatant were determined with an enzyme-linked immunosorbent assay (ELISA) kit following the manufacturer’s instructions.

Techniques: Control, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Biomarker Discovery

Journal: International immunopharmacology

Article Title: MiR-625-5p is a potential therapeutic target in sepsis by regulating CXCL16/CXCR6 axis and endothelial barrier.

doi: 10.1016/j.intimp.2024.112508

Figure Lengend Snippet: Fig. 3. miR-625-5p regulated CXCL16 transcription and expression. EA.hy926 cells were transfected with miR-625-5p mimic (100 nM) or negative control (miR- 625-5p mimic NC) for 48 h. qRT-PCR analysis of miRNA expression levels for miR-625-5p by transfection of miR-625-5p mimics (a). The relative level of CXCL16 was detected by qRT-PCR (b), CXCL16 supernatant levels were quantified by ELISA (c), whereas the expression levels in EA.hy926 cells were detected by Western blotting (d). Quantification of CXCL16 is illustrated in (e). *P < 0.05, **P < 0.01.

Article Snippet: Serum levels of syndecan-1 (EK1339, BOSTER, China), heparan sulfate (356350, Usbiological, USA), and VE-cadherin (DCADV0, R&D Systems, USA), and CXCL16 (EK0741, BOSTER, China) levels in the serum and cell supernatant were determined with an enzyme-linked immunosorbent assay (ELISA) kit following the manufacturer’s instructions.

Techniques: Expressing, Transfection, Negative Control, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Western Blot

Journal: International immunopharmacology

Article Title: MiR-625-5p is a potential therapeutic target in sepsis by regulating CXCL16/CXCR6 axis and endothelial barrier.

doi: 10.1016/j.intimp.2024.112508

Figure Lengend Snippet: Fig. 4. Inhibitor of miR-625-5p attenuated LPS-induced EA.hy926 cell barrier injury. EA.hy926 cells were transfected with the miR-625-5p inhibitor (50 nM) or negative control (micrOFF inhibitor NC) for 48 h and then exposed to LPS (5 and 10 µg/mL) for 6 h. The relative level of CXCL16 was detected by qRT-PCR (a), the expression levels of CXCL16 in the EA.hy926 cells were detected by Western blotting (b), and CXCL16 supernatant levels were quantified by ELISA (c). Quantification of CXCL16 is illustrated in (d). The effect of LPS and miR-625-5p mimic on the permeability of EA.hy926 cell were assessed using FITC-dextran and TEER methods (e, f). *P < 0.05, **P < 0.01.

Article Snippet: Serum levels of syndecan-1 (EK1339, BOSTER, China), heparan sulfate (356350, Usbiological, USA), and VE-cadherin (DCADV0, R&D Systems, USA), and CXCL16 (EK0741, BOSTER, China) levels in the serum and cell supernatant were determined with an enzyme-linked immunosorbent assay (ELISA) kit following the manufacturer’s instructions.

Techniques: Transfection, Negative Control, Quantitative RT-PCR, Expressing, Western Blot, Enzyme-linked Immunosorbent Assay, Permeability

Journal: International immunopharmacology

Article Title: MiR-625-5p is a potential therapeutic target in sepsis by regulating CXCL16/CXCR6 axis and endothelial barrier.

doi: 10.1016/j.intimp.2024.112508

Figure Lengend Snippet: Fig. 6. CXCL16 knockdown inhibited LPS-induced endothelial cell injury in vitro. EA.hy926 cells were transfected with CXCL16 siRNA (siCXCL16-1, 2, or 3) or negative control (siRNA NC) and CXCL16 expression was determined after 24 h. The relative level of CXCL16 were detected by qRT-PCR (a). EA.hy926 cells were divided into four groups: Group 1, control; Group 2, LPS; Group 3, LPS + siCXCL16; Group 4, siRNA NC. The cells in Group 3 were transfected with CXCL16 siRNA (siCXCL16), whereas cells in the other groups were transfected with negative control siRNA (siRNA NC). At 24 h post-transfection, the cells in Groups 2 and 3 were treated with 10 µg/mL LPS. Endothelial permeability was measured by using FITC- dextran and TEER 6 h after LPS treatment (b, c). The levels of HS, SDC-1, claudin5, occludin, and VE-cadherin were detected by Western blotting (d). Quantification of HS, SDC-1, claudin5, occludin, and VE-cadherin are shown in (e): *P < 0.05, vs. Control group; & P < 0.05, vs. LPS group. Immunofluorescence images of HS and SDC-1 in EA.hy926 cells (f, h; magnification, ×200; scale bar, 150 µm). Fluo rescence intensity analysis of g and i. *P < 0.05, **P < 0.01.

Article Snippet: Serum levels of syndecan-1 (EK1339, BOSTER, China), heparan sulfate (356350, Usbiological, USA), and VE-cadherin (DCADV0, R&D Systems, USA), and CXCL16 (EK0741, BOSTER, China) levels in the serum and cell supernatant were determined with an enzyme-linked immunosorbent assay (ELISA) kit following the manufacturer’s instructions.

Techniques: Knockdown, In Vitro, Transfection, Negative Control, Expressing, Quantitative RT-PCR, Control, Permeability, Western Blot, Immunofluorescence

Journal: International immunopharmacology

Article Title: MiR-625-5p is a potential therapeutic target in sepsis by regulating CXCL16/CXCR6 axis and endothelial barrier.

doi: 10.1016/j.intimp.2024.112508

Figure Lengend Snippet: Fig. 9. Mechanisms by which miR-625-5p disrupts lung endothelial barrier integrity. miR-625-5p level may be an effective biomarker for predicting 28-day mortality in patients with sepsis or septic shock. Furthermore, LPS-induced vascular endothelial hyper-permeability by regulating miR-625-5p/CXCL16/CXCR6 axis in sepsis. ① LPS increased miR‑625-5p expression in EA.hy926 cells; ② miR-625-5p positively regulated CXCL16 in EA.hy926 cells; ③ Treatment of EA.hy926 cells with LPS significantly increased CXCL16 release; ④ CXCL16 combines with CXCR6; ⑤ CXCR6 mediated the effects of CXCL16 on endothelial barrier dysfunction.

Article Snippet: Serum levels of syndecan-1 (EK1339, BOSTER, China), heparan sulfate (356350, Usbiological, USA), and VE-cadherin (DCADV0, R&D Systems, USA), and CXCL16 (EK0741, BOSTER, China) levels in the serum and cell supernatant were determined with an enzyme-linked immunosorbent assay (ELISA) kit following the manufacturer’s instructions.

Techniques: Biomarker Discovery, Permeability, Expressing

Journal: Genome Medicine

Article Title: Single-cell RNA transcriptome analysis of CNS immune cells reveals CXCL16/CXCR6 as maintenance factors for tissue-resident T cells that drive synapse elimination

doi: 10.1186/s13073-022-01111-0

Figure Lengend Snippet: CXCL16 levels increase on IBA1 + cells during acute infection and return to baseline levels during recovery. a Representative microscopy images of RNA in situ/immunohistochemistry (ISH/IHC) for Cxcl16 and IBA1 in the cortex of mock- or WNV-infected mice at 7, 25, ad 52 DPI. The middle panel is the inset of the white box in 7 DPI. b – d Quantification of the ISH/IHC for Cxcl16 + area ( b ); IBA1 + Cxcl16 + area normalized to the total IBA1 + area, represented as fold change over mock ( c ); or IBA1 + Cxcl16 + area normalized to the total Cxcl16 + area ( d ) in the cortex or hippocampus of mock- or WNV-infected mice at 7, 25, and 52 DPI. e ELISA for CXCL16 in the cortex, hippocampus, cervical lymph nodes, meninges, and blood at 7 and 25 DPI. f Representative immunostaining of the cortex for CXCR6-GFP, CD3, IBA1, and DAPI. Scale bars, 50 μm. Data represent the mean±s.e.m. and were analyzed by one-way ANOVA or unpaired Student’s t -test. * P <0.05, ** P <0.005, *** P < 0.001, **** P < 0.0001

Article Snippet: The level of CXCL16 was detected using a commercial ELISA kit (DY503, R&D Systems, Inc., MN, USA), according to the manufacturer’s guidelines.

Techniques: Infection, Microscopy, In Situ, Immunohistochemistry, Enzyme-linked Immunosorbent Assay, Immunostaining

Journal:

Article Title: Expression and regulation of the chemokine CXCL16 in Crohn's disease and models of intestinal inflammation

doi: 10.1002/ibd.21306

Figure Lengend Snippet: CXCL16 and CXCR6 mRNA expression in inflamed and non-inflamed colonic lesions from patients with CD determined by quantitative RT-PCR and normalized to beta-actin expression levels. The current medical therapy during biopsy sampling and the anatomic site, from which the samples were taken, are given for all patients.

Article Snippet: For the quantification of CXCL16 in serum samples of IBD patients and healthy controls as well as in cell culture supernatants, human CXCL16 Quantikine Elisa Kit (R&D Systems, Minneapolis, MN) was used according to the manufacturer’s instructions.

Techniques: Expressing, Quantitative RT-PCR, Sampling

Journal:

Article Title: Expression and regulation of the chemokine CXCL16 in Crohn's disease and models of intestinal inflammation

doi: 10.1002/ibd.21306

Figure Lengend Snippet: (A) Northern Blot analysis of mRNA derived from murine tissue of two wt BALB/c mice as indicated. Note the inverse segment-specific expression of CXCL16 and CXCR6 mRNA in the murine gastrointestinal tract with high CXCL16 expression in the small intestine and high CXCR6 expression in the colon. (B) CXCL16 and CXCR6 mRNA expression in human IEC lines and the murine IEC line CMT93 were analyzed by RT-PCR. (C) Western blot analysis of CXCR6 protein expression in cytosolic (C) and membrane (M) protein fractions of HT-29 cells. (D) Immunocytochemical staining reveals a membrane associated expression of CXCR6 in HT-29 cells (anti-CXCR6 antibody detected with a FITC conjugated secondary antibody, nucleus: stained with Hoechst 33342). (E) No CXCR6 specific staining was obtained using the secondary antibody (FITC conjugated) only. (F) Western blot analysis of CXCL16 protein expression in cytosolic (C) and membrane (M) protein fractions of HT-29 cells. (G) Immunocytochemical staining with a CXCL16 specific antibody in HT-29 cells. (H) No CXCL16 specific staining was detected using the secondary antibody only.

Article Snippet: For the quantification of CXCL16 in serum samples of IBD patients and healthy controls as well as in cell culture supernatants, human CXCL16 Quantikine Elisa Kit (R&D Systems, Minneapolis, MN) was used according to the manufacturer’s instructions.

Techniques: Northern Blot, Derivative Assay, Expressing, Reverse Transcription Polymerase Chain Reaction, Western Blot, Membrane, Staining

Journal:

Article Title: Expression and regulation of the chemokine CXCL16 in Crohn's disease and models of intestinal inflammation

doi: 10.1002/ibd.21306

Figure Lengend Snippet: (A) Detection of phosphorylated ERK-1/2 in CMT93 IEC lines after CXCL16 stimulation (100 ng/mL). (B) Phospho-ERK1/2 activation by CXCL16 in CMT93 cells pretreated with the MEK-1 inhibitor PD98059 (5 μmol/L) and the PI3 kinase inhibitor wortmannin (200 nmol/L). (C) Phosphorylated SAPK/JNK kinases in CMT93 cells after CXCL16 stimulation (100 ng/mL). (D) CXCL16 stimulation of CMT93 cells resulted also in very weak phosphorylation of p38. (E) Phosphorylated Akt in CXCL16 stimulated CMT93 cells (including wortmannin pretreated cells) and in (F) PD98059 pretreated cells. (G) ERK-1/2 MAP kinases are activated in HT-29 cells following CXCL16 stimulation. (H) CXCL16 leads to Akt phosphorylation in HT-29 cells. For all signaling experiments (Fig. 2A–H), one representative experiment of three performed is shown.

Article Snippet: For the quantification of CXCL16 in serum samples of IBD patients and healthy controls as well as in cell culture supernatants, human CXCL16 Quantikine Elisa Kit (R&D Systems, Minneapolis, MN) was used according to the manufacturer’s instructions.

Techniques: Activation Assay, Phospho-proteomics

Journal:

Article Title: Expression and regulation of the chemokine CXCL16 in Crohn's disease and models of intestinal inflammation

doi: 10.1002/ibd.21306

Figure Lengend Snippet: (A) The proinflammatory cytokines TNF-α (50 μg/mL), IL-1β (10 μg/mL) and IFN-γ (1000 U/mL) significantly increase expression of CXCL16 mRNA in HT-29 cells as determined by quantitative PCR (*p<0.005 vs. t=0h) while LPS (1 μg/mL) has no stimulating effect. (B) Similarly, we measured soluble CXCL16 protein concentration by ELISA cell culture supernatants of HT-29 cells during cytokine stimulation (*p<0.05 vs. t = 0 h). Note the corresponding but delayed kinetics of CXCL16 protein release in comparison to the CXCL16 mRNA increase. The detection limit of the ELISA was 156 pg/mL (C) CXCL16 mRNA expression is increased 3.5-fold in ileal epithelial cells from TNFΔARE heterozygous mice (n=9) compared to wt mice (n=9) as determined by quantitative PCR (*p<0.05 vs. wt). Additionally, there was a higher CXCL16 expression in ileal and jejunal epithelial cells compared to colonic epithelial cells (data not shown). (D) CXCL16 mRNA expression is increased in colonic tissue from 1×106 pfu MCMV i.v. infected C57BL/6 mice (n=11) in comparison to PBS injected C57BL/6 control mice (n=4, * p=0.03 vs. controls).

Article Snippet: For the quantification of CXCL16 in serum samples of IBD patients and healthy controls as well as in cell culture supernatants, human CXCL16 Quantikine Elisa Kit (R&D Systems, Minneapolis, MN) was used according to the manufacturer’s instructions.

Techniques: Expressing, Real-time Polymerase Chain Reaction, Protein Concentration, Enzyme-linked Immunosorbent Assay, Cell Culture, Comparison, Infection, Injection, Control

Journal:

Article Title: Expression and regulation of the chemokine CXCL16 in Crohn's disease and models of intestinal inflammation

doi: 10.1002/ibd.21306

Figure Lengend Snippet: Immunohistochemical analysis of biopsies from three different CD patients (A–D: patient 1, E–H: patient 2, I–L: patient 3) reveals higher mucosal infiltration with CXCL16-expressing immune cells in inflamed regions (C/D; G/H; K/L) in comparison to uninflamed tissue (A/B; E/F; I/J). 20×, 40×: magnification.

Article Snippet: For the quantification of CXCL16 in serum samples of IBD patients and healthy controls as well as in cell culture supernatants, human CXCL16 Quantikine Elisa Kit (R&D Systems, Minneapolis, MN) was used according to the manufacturer’s instructions.

Techniques: Immunohistochemical staining, Expressing, Comparison

Journal:

Article Title: Expression and regulation of the chemokine CXCL16 in Crohn's disease and models of intestinal inflammation

doi: 10.1002/ibd.21306

Figure Lengend Snippet: (A) In the sera of patients with Crohn’s disease (n=30), significantly higher CXCL16 serum protein levels were detected in comparison to healthy controls (n=30; * p<0.01 vs. controls; UC: n=17; p=0.057 vs. controls). (B) CXCL16 expression was particularly high in CD patients with active disease (CDAI >150; * p=0.02 vs. controls).

Article Snippet: For the quantification of CXCL16 in serum samples of IBD patients and healthy controls as well as in cell culture supernatants, human CXCL16 Quantikine Elisa Kit (R&D Systems, Minneapolis, MN) was used according to the manufacturer’s instructions.

Techniques: Comparison, Expressing

Journal: PLoS ONE

Article Title: Urinary CXCL1: A Novel Predictor of IgA Nephropathy Progression

doi: 10.1371/journal.pone.0119033

Figure Lengend Snippet: Baseline clinical and laboratory data and levels of urinary CXCL1 in patients with IgAN.

Article Snippet: Urinary CXCL1 were quantified by a standard sandwich ELISA assays using the DuoSet human CXCL1 ELISA kits (R&D Systems, Minneapolis, MN, USA), according to the manufacturer’s protocol.

Techniques:

Journal: PLoS ONE

Article Title: Urinary CXCL1: A Novel Predictor of IgA Nephropathy Progression

doi: 10.1371/journal.pone.0119033

Figure Lengend Snippet: Urinary CXCL1 was calibrated against urine creatinine before the levels were compared. The urinary CXCL1 levels in patients with IgAN, MCD, MN, FSGS and LN were significantly higher than those in healthy controls (0, IQR 0–0, P<0.001 in all glomerulonephritis). Urinary CXCL1 levels were significantly higher in patients with IgAN (18.29 pg/mg, IQR 10.17–33.47 pg/mg) than in MCD (3.03 pg/mg, IQR 0–11.28 pg/mg, P<0.001), MN (5.89 pg/mg, IQR 0–23.54 pg/mg, P<0.001) and FSGS (6.10 pg/mg, IQR 0–17.53 pg/mg, P<0.001), while no significant difference was observed between patient with IgAN and LN (18.29 pg/mg, IQR 10.17–33.47 pg/mg vs. 23.12 pg/mg, IQR 7.41–48.17 pg/mg; P = 0.873). (*, significant difference between IgAN group and other GN groups; **, significant difference between healthy control group and disease groups).

Article Snippet: Urinary CXCL1 were quantified by a standard sandwich ELISA assays using the DuoSet human CXCL1 ELISA kits (R&D Systems, Minneapolis, MN, USA), according to the manufacturer’s protocol.

Techniques: Control

Journal: PLoS ONE

Article Title: Urinary CXCL1: A Novel Predictor of IgA Nephropathy Progression

doi: 10.1371/journal.pone.0119033

Figure Lengend Snippet: Multivariable linear regression analyses for urinary CXCL1 level.

Article Snippet: Urinary CXCL1 were quantified by a standard sandwich ELISA assays using the DuoSet human CXCL1 ELISA kits (R&D Systems, Minneapolis, MN, USA), according to the manufacturer’s protocol.

Techniques:

Journal: PLoS ONE

Article Title: Urinary CXCL1: A Novel Predictor of IgA Nephropathy Progression

doi: 10.1371/journal.pone.0119033

Figure Lengend Snippet: For patients with high baseline urinary CXCL1 levels (case 1 & case 2), the variation tendencies of urinary CXCL1 and proteinuria were almost the same (A & B). For patients with middle levels of baseline urinary CXCL1 (case 3 & case 4), although proteinuria relieved a lot, urinary CXCL1 levels just decreased a little (C & D). For patients with low levels of baseline urinary CXCL1 (case 5 & case 6), decreased urinary CXCL1 levels were not observed accompanied with proteinuria remission (E & F). Treatments are depicted as grey-level-coded bars, on behalf of the period of using time.

Article Snippet: Urinary CXCL1 were quantified by a standard sandwich ELISA assays using the DuoSet human CXCL1 ELISA kits (R&D Systems, Minneapolis, MN, USA), according to the manufacturer’s protocol.

Techniques:

Journal: PLoS ONE

Article Title: Urinary CXCL1: A Novel Predictor of IgA Nephropathy Progression

doi: 10.1371/journal.pone.0119033

Figure Lengend Snippet: The variation tendency of urinary CXCL1 was not the same with that of SBP, not only in patients with high baseline urinary CXCL1 levels (case 1 & case 2, A & B), but also in patients with middle baseline urinary CXCL1 levels (case 3 & case 4, C & D) and patients with low baseline urinary CXCL1 levels (case 5 & case 6, E & F). Treatments are depicted as grey-level-coded bars, on behalf of the period of using time.

Article Snippet: Urinary CXCL1 were quantified by a standard sandwich ELISA assays using the DuoSet human CXCL1 ELISA kits (R&D Systems, Minneapolis, MN, USA), according to the manufacturer’s protocol.

Techniques:

Journal: PLoS ONE

Article Title: Urinary CXCL1: A Novel Predictor of IgA Nephropathy Progression

doi: 10.1371/journal.pone.0119033

Figure Lengend Snippet: Clinical outcomes according to urinary CXCL1 level.

Article Snippet: Urinary CXCL1 were quantified by a standard sandwich ELISA assays using the DuoSet human CXCL1 ELISA kits (R&D Systems, Minneapolis, MN, USA), according to the manufacturer’s protocol.

Techniques:

Journal: PLoS ONE

Article Title: Urinary CXCL1: A Novel Predictor of IgA Nephropathy Progression

doi: 10.1371/journal.pone.0119033

Figure Lengend Snippet: Patients with IgAN were divided into two groups, according to the median urinary CXCL1 level (18.29 pg/mg). Those with CXCL1 levels less than 18.29 pg/mg were classified as group A, while others (CXCL1 levels above 18.29 pg/mg) were classified as group B. IgAN patients in group B had significantly lower renal survival rate than those in group A (P < 0.001). The renal survival at first and fifth year for patients in group A were 100.0% and 96.3%, while for patients in group B were 97.2% and 81.3% (Log Rank test, p<0.001).

Article Snippet: Urinary CXCL1 were quantified by a standard sandwich ELISA assays using the DuoSet human CXCL1 ELISA kits (R&D Systems, Minneapolis, MN, USA), according to the manufacturer’s protocol.

Techniques:

Journal: PLoS ONE

Article Title: Urinary CXCL1: A Novel Predictor of IgA Nephropathy Progression

doi: 10.1371/journal.pone.0119033

Figure Lengend Snippet: Risks of composite end-point of natural log–transformed CXCL1.

Article Snippet: Urinary CXCL1 were quantified by a standard sandwich ELISA assays using the DuoSet human CXCL1 ELISA kits (R&D Systems, Minneapolis, MN, USA), according to the manufacturer’s protocol.

Techniques:

Journal: PLoS ONE

Article Title: Urinary CXCL1: A Novel Predictor of IgA Nephropathy Progression

doi: 10.1371/journal.pone.0119033

Figure Lengend Snippet: Using composite outcome at 24 (solid line), 48 (dashed line), and 72 months (dotted line) as status variable, respectively, the areas under the ROC curve (AUC) for proteinuria (A), reciprocal of estimated glomerular filtration rate (1/eGFR) (B) and urinary CXCL1 (C) were comparable (proteinuria: 0.720 (24 mo), 0.690 (48 mo), 0.612 (72 mo); 1/eGFR: 0.826 (24 mo), 0.809 (48 mo), and 0.690 (72 mo); urinary CXCL1: 0.770 (24 mo), 0.651 (48 mo), 0.668 (72 mo)). When combined urinary CXCL1 level with proteinuria and 1/eGFR (solid line), the AUCs were higher than proteinuria alone (dotted line) or proteinuria plus 1/eGFR (dashed line) (D).

Article Snippet: Urinary CXCL1 were quantified by a standard sandwich ELISA assays using the DuoSet human CXCL1 ELISA kits (R&D Systems, Minneapolis, MN, USA), according to the manufacturer’s protocol.

Techniques: Filtration

Journal: PLoS ONE

Article Title: Urinary CXCL1: A Novel Predictor of IgA Nephropathy Progression

doi: 10.1371/journal.pone.0119033

Figure Lengend Snippet: Comparison of two models of survival analysis.

Article Snippet: Urinary CXCL1 were quantified by a standard sandwich ELISA assays using the DuoSet human CXCL1 ELISA kits (R&D Systems, Minneapolis, MN, USA), according to the manufacturer’s protocol.

Techniques: Comparison

Journal: bioRxiv

Article Title: CXCR6⁺ natural killer cell immunotherapy preserves CD4⁺ T helper cells in humanized mice

doi: 10.64898/2026.05.28.728486

Figure Lengend Snippet: A) Expansion kinetics of sorted CXCR6⁻ and CXCR6⁺ NK cells from independent peripheral blood NK-cell donors during 15 days of ex vivo culture with CXCL16-expressing feeder cells, irradiated allogeneic PBMC feeders, and cytokine supplementation. B) Representative post-expansion flow cytometry plots showing maintenance of CXCR6-defined NK-cell phenotypes following expansion. C) Representative intracellular HIV-Gag flow cytometry plots from the indicated culture conditions. Activated primary CD4⁺ T cells from a single peripheral blood donor were infected with HIV-1 Q23.17 at an MOI of 0.01 and cultured alone or co-cultured for 3 days with day 14-expanded CXCR6⁺ or CXCR6⁻ NK cells at a 1:1 effector-to-target ratio. HIV infection was quantified by intracellular HIV-Gag staining and flow cytometry. D) Quantification of HIV suppression assay results. CXCR6⁺ and CXCR6⁻ NK cells were tested using NK cells from five independent genetically unrelated donors, with the same HIV-infected CD4⁺ T-cell target donor used across all NK donor and subset conditions. For each NK donor and condition, the CD4 + T cell-frequency was determined by flow cytometry, triplicate technical wells per donor were averaged before statistical analysis, and the five NK donors were analyzed as biological replicates. Donor-level paired comparisons were analyzed using two-sided paired tests with Holm correction for multiple comparisons. CXCR6⁺ NK cells significantly reduced the frequency of HIV-Gag⁺ CD4⁺ T cells compared with HIV-only cultures, p = 0.000064. CXCR6⁻ NK cells also significantly reduced HIV-Gag⁺ CD4⁺ T cells compared with HIV-only cultures, p = 0.004059. CXCR6⁺ NK cells suppressed HIV significantly more effectively than paired CXCR6⁻ NK cells, p = 0.009637. Data are presented as mean ± SD, with each color representing an independent NK-cell donor. **p < 0.01, ****p < 0.0001.

Article Snippet: RPMI 8866 cells were transduced with lentiviral particles (pLenti C mGFP, Origene) carrying CXCL16 transcript variant 1 (TV1, NM_022059) or variant 2 (TV2, NM_001100812).

Techniques: Ex Vivo, Expressing, Irradiation, Flow Cytometry, Infection, Cell Culture, Staining, HIV Inhibition Assay

Journal: Neoplasia (New York, N.Y.)

Article Title: High metastatic tumor-derived CXCL16 mediates liver colonization metastasis by inducing Kupffer cell polarization via the PI3K/AKT/FOXO3a pathway

doi: 10.1016/j.neo.2025.101174

Figure Lengend Snippet: CXCL16 mediate the intercommunication between CRC cells and KCs. (A)RNA-seq gene expression heatmap in CT26 and CT26-LM group. (B)RNA-seq gene expression heatmap in CT26 and co-CT26 group. (C)Venn diagram of genes positively correlated with liver metastasis analyzed through transcriptome microarrays between parent CT26 cells and CT26-LM or CT26 and co-CT26 cells. (D)Heatmap of top 20 changed genes among parent CT26 cells, LM and co-cultured CT26 cells. (E)Heatmap of top 10 changed cytokine genes among parent CT26 cells, LM and co-cultured CT26 cells. (F)qRT-PCR analysis of the mRNA expression of CXCL14, IL24, CXCL16 and IL1A in CT26 and CT26-LM cells. (G)qRT-PCR analysis of the mRNA expression of CXCL14, IL24, CXCL16 and IL1A in CT26 and co-CT26 cells. (H)ELISA assay revealed the CXCL16 upregulated in culture supernatants from CT26-LM cells. (I)Western blot analysis showed the changes of CXCL16 protein level in CT26, CT26-LM and co-CT26 cells.

Article Snippet: CXCL16 levels in cell culture media and mouse serum were quantified using a Mouse CXCL16 ELISA kit (Elabscience, China) according to the manufacturer's protocol.

Techniques: RNA Sequencing, Gene Expression, Cell Culture, Quantitative RT-PCR, Expressing, Enzyme-linked Immunosorbent Assay, Western Blot

Journal: Neoplasia (New York, N.Y.)

Article Title: High metastatic tumor-derived CXCL16 mediates liver colonization metastasis by inducing Kupffer cell polarization via the PI3K/AKT/FOXO3a pathway

doi: 10.1016/j.neo.2025.101174

Figure Lengend Snippet: CXCL16/PI3K/FOXO3a signaling pathway mediates M2 polarization of KCs. (A)Western blot showed the protein expression of M2 markers in different concentration of CXCL16. (B)Western blot showed the protein expression of M2 markers in different concentration of ML339. (C)Western blot analysis of M2 markers (CD206, ARG1) and PI3K/AKT1/FOXO3a pathway in ImKC when treated with CXCL16 (200ng/ml), LM-CM or ML339. (D)Western blot analysis of M2 markers (CD206, ARG1) and PI3K/AKT1/FOXO3a pathway in ImKC when treated with CXCL16 (200ng/ml), LM-CM or LY294002. (E)Identification of transcription factor FOXO3a binding site. (F)Prediction of FOXO3a binding sites in the CD206 promoter region using JASPAR. (G)The promoter sequence of CD206 contains 3 putative FOXO3a transcriptional factor binding sites. (H)Luciferase reporter assays for the activity of wild-type and mutant CD206 promoters. (I)Quantitative analysis of ChIP experiments performed on DNA samples precipitated with antibodies against FOXO3a and IgG using primers detecting FOXO3a binding sites on CD206 promoter.

Article Snippet: CXCL16 levels in cell culture media and mouse serum were quantified using a Mouse CXCL16 ELISA kit (Elabscience, China) according to the manufacturer's protocol.

Techniques: Western Blot, Expressing, Concentration Assay, Binding Assay, Sequencing, Luciferase, Activity Assay, Mutagenesis

Journal: Neoplasia (New York, N.Y.)

Article Title: High metastatic tumor-derived CXCL16 mediates liver colonization metastasis by inducing Kupffer cell polarization via the PI3K/AKT/FOXO3a pathway

doi: 10.1016/j.neo.2025.101174

Figure Lengend Snippet: Intervention of CXCL16/CXCR6 for suppression of CT26-LM metastasis in vivo. (A)Schematic representation of BALB/c mice were treated with ML339 once every two days after spleen injection CT26. (B)Liver metastasis nodules between control(DMSO) group and ML339 group (metastatic tumor nodules shown by arrows). (C)Dotplot showed the number of metastatic tumors between control(DMSO) group and ML339 group. (D)Barplot showed the whole liver weight between control(DMSO) group and ML339 group. (E)F4/80 IHC staining and quantitative analysis of AOD in tumor; scale bar 50μm. (F)Relative mRNA expression of M2 markers and M1 markers in control(DMSO) and ML339 group liver tissue. (G-H)IF staining of ARG1 and CD206 in control(DMSO) and ML339 group liver tissue; scale bar 50μm.

Article Snippet: CXCL16 levels in cell culture media and mouse serum were quantified using a Mouse CXCL16 ELISA kit (Elabscience, China) according to the manufacturer's protocol.

Techniques: In Vivo, Injection, Control, Immunohistochemistry, Expressing, Staining

Journal: Neoplasia (New York, N.Y.)

Article Title: High metastatic tumor-derived CXCL16 mediates liver colonization metastasis by inducing Kupffer cell polarization via the PI3K/AKT/FOXO3a pathway

doi: 10.1016/j.neo.2025.101174

Figure Lengend Snippet: Diagrammatic sketch for CXCL16/PI3K/AKT1/FOXO3a activation in M2-KCs to promote MET of liver-metastatic cells.

Article Snippet: CXCL16 levels in cell culture media and mouse serum were quantified using a Mouse CXCL16 ELISA kit (Elabscience, China) according to the manufacturer's protocol.

Techniques: Activation Assay