Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Human trophoblasts recruited T lymphocytes and monocytes into decidua by secretion of chemokine CXCL16 and interaction with CXCR6 in the first-trimester pregnancy.

doi: 10.4049/jimmunol.180.4.2367

Figure Lengend Snippet: FIGURE 1. The transcription and translation of chemokine CXCL16 in human first-trimester trophoblasts. The real-time quantitative PCR was used to analyze transcription of CXCL16 in trophoblasts and JAR cells with cyclophilin A (CyP) as positive control. Three independent experiments were done (including 15 placental samples) and the results were reproducible (A). The specific brown-colored stainings for CXCL16 were recognized in the cytoplasm and cytomembrane of the primary-cultured villous cytotrophoblasts (B), and extravillous cytotrophoblasts (C) by immunocytochemistry, and the positive stainings were recognized by villous cytotrophoblasts and ST (E) and by extravillous cytotrophoblasts (F) by immunohistochemistry. No background staining was observed in goat isotype controls (D and G). The experiments were repeated five times with five placenta samples, respectively. The picture is a representative one. Magnification: B, D, E, and G, 200; C and F, 400.

Article Snippet: Anti-human CXCL16 Ab (R&D Systems) were administrated to detect whether cytotrophoblasts express CXCL16.

Techniques: Real-time Polymerase Chain Reaction, Positive Control, Cell Culture, Immunocytochemistry, Immunohistochemistry, Staining

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Human trophoblasts recruited T lymphocytes and monocytes into decidua by secretion of chemokine CXCL16 and interaction with CXCR6 in the first-trimester pregnancy.

doi: 10.4049/jimmunol.180.4.2367

Figure Lengend Snippet: FIGURE 2. The accumulated concentration of CXCL16 in culture me- dium of first-trimester cytotrophoblasts was examined by ELISA. Purified cytotrophoblasts were seeded at 1 106, 5 105, 1 105, 5 104, 1 104, and 5 103 cells/ml, and then the supernatants were collected and measured after 12, 24, 36, 48, 60, 72, and 100 h of culture. The results showed CXCL16 accumulation in the medium during the course of culture. Each point represents the mean SD obtained from duplicate dishes, and the data are representative of two independent experiments.

Article Snippet: Anti-human CXCL16 Ab (R&D Systems) were administrated to detect whether cytotrophoblasts express CXCL16.

Techniques: Concentration Assay, Enzyme-linked Immunosorbent Assay

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Human trophoblasts recruited T lymphocytes and monocytes into decidua by secretion of chemokine CXCL16 and interaction with CXCR6 in the first-trimester pregnancy.

doi: 10.4049/jimmunol.180.4.2367

Figure Lengend Snippet: FIGURE 6. Correlation of CXCL16 chemotaxis to CXCR6 expression level in the peripheral (A) and decidual (B) immune cells. CXCL16 che- motaxis was shown as the ratio of the number of migrated immune cells treated by CXCL16 (100 ng/ml) to the number of migrated immune cells in the control.

Article Snippet: Anti-human CXCL16 Ab (R&D Systems) were administrated to detect whether cytotrophoblasts express CXCL16.

Techniques: Chemotaxis Assay, Expressing, Control

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Human trophoblasts recruited T lymphocytes and monocytes into decidua by secretion of chemokine CXCL16 and interaction with CXCR6 in the first-trimester pregnancy.

doi: 10.4049/jimmunol.180.4.2367

Figure Lengend Snippet: FIGURE 1. The transcription and translation of chemokine CXCL16 in human first-trimester trophoblasts. The real-time quantitative PCR was used to analyze transcription of CXCL16 in trophoblasts and JAR cells with cyclophilin A (CyP) as positive control. Three independent experiments were done (including 15 placental samples) and the results were reproducible (A). The specific brown-colored stainings for CXCL16 were recognized in the cytoplasm and cytomembrane of the primary-cultured villous cytotrophoblasts (B), and extravillous cytotrophoblasts (C) by immunocytochemistry, and the positive stainings were recognized by villous cytotrophoblasts and ST (E) and by extravillous cytotrophoblasts (F) by immunohistochemistry. No background staining was observed in goat isotype controls (D and G). The experiments were repeated five times with five placenta samples, respectively. The picture is a representative one. Magnification: B, D, E, and G, 200; C and F, 400.

Article Snippet: The human CXCL16 ELISA kit (R&D Systems) was used to measure chemokine production in each supernatant and CM according to the manufacturer’s instructions.

Techniques: Real-time Polymerase Chain Reaction, Positive Control, Cell Culture, Immunocytochemistry, Immunohistochemistry, Staining

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Human trophoblasts recruited T lymphocytes and monocytes into decidua by secretion of chemokine CXCL16 and interaction with CXCR6 in the first-trimester pregnancy.

doi: 10.4049/jimmunol.180.4.2367

Figure Lengend Snippet: FIGURE 2. The accumulated concentration of CXCL16 in culture me- dium of first-trimester cytotrophoblasts was examined by ELISA. Purified cytotrophoblasts were seeded at 1 106, 5 105, 1 105, 5 104, 1 104, and 5 103 cells/ml, and then the supernatants were collected and measured after 12, 24, 36, 48, 60, 72, and 100 h of culture. The results showed CXCL16 accumulation in the medium during the course of culture. Each point represents the mean SD obtained from duplicate dishes, and the data are representative of two independent experiments.

Article Snippet: The human CXCL16 ELISA kit (R&D Systems) was used to measure chemokine production in each supernatant and CM according to the manufacturer’s instructions.

Techniques: Concentration Assay, Enzyme-linked Immunosorbent Assay

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Human trophoblasts recruited T lymphocytes and monocytes into decidua by secretion of chemokine CXCL16 and interaction with CXCR6 in the first-trimester pregnancy.

doi: 10.4049/jimmunol.180.4.2367

Figure Lengend Snippet: FIGURE 6. Correlation of CXCL16 chemotaxis to CXCR6 expression level in the peripheral (A) and decidual (B) immune cells. CXCL16 che- motaxis was shown as the ratio of the number of migrated immune cells treated by CXCL16 (100 ng/ml) to the number of migrated immune cells in the control.

Article Snippet: The human CXCL16 ELISA kit (R&D Systems) was used to measure chemokine production in each supernatant and CM according to the manufacturer’s instructions.

Techniques: Chemotaxis Assay, Expressing, Control

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: Proceedings of the National Academy of Sciences of the United States of America

Article Title: CXCR4 inhibition in human pancreatic and colorectal cancers induces an integrated immune response

doi: 10.1073/pnas.2013644117

Figure Lengend Snippet: The effect of CXCL12-stimulated CXCR4 on chemokine receptor-mediated migration of human immune cells. ( Left ) The coexpression of CXCR4 with ( A ) CXCR1, ( B ) CXCR3, ( C ) CXCR5, ( D ) CXCR6, and ( E ) CCR2 on human immune cell lines was evaluated by flow cytometry after staining with antibodies specific for the relevant chemokine receptors. Gray peaks indicate isotype controls. ( Center ) The effect of CXCL12-stimulation of CXCR4 on the chemotactic responses of A CXCR1-coexpressing Jurkat T lymphoblastoid cells to CXCL8, ( B ) CXCR3-coexpressing HSB2DP T lymphoblastoid cells to CXCL10, ( C ) CXCR5-coexpressing Raji B lymphoblastoid cells to CXCL13, ( D ) CXCR6-coexpressing Jurkat T lymphoblastoid cells to CXCL16, and ( E ) CCR2-coexpressing Molm13 monocytoid cells to CCL2 was assessed by including CXCL12 in the upper chamber (blue) and the other chemokines in the lower chamber (red) in the Boyden two-chamber assay. ( Right ) The chemotaxis assays were performed with the five cell lines when the placement of the chemokines in the Boyden chambers was reversed. Bar diagrams display mean and SEM (n = 3-4). Statistical analysis by Student’s t test: *** P < 0.001; **** P < 0.0001; ns, not significant.

Article Snippet: The concentration of each chemokine used is as following: CXCL8 (R&D Systems, 208-IL), 20 ng/mL; CXCL10 (R&D Systems, 266-IP), 1,000 ng/mL; CXCL13 (R&D Systems, 801-CX), 1,000 ng/mL; CXCL16 (R&D Systems, 976-CX), 50 ng/mL; CCL2 (R&D Systems, 279-MC), 200 ng/mL

Techniques: Migration, Flow Cytometry, Staining, Boyden Chamber Assay, Chemotaxis Assay

Journal: The Journal of investigative dermatology

Article Title: Constitutive expression and regulated release of the transmembrane chemokine CXCL16 in human and murine skin.

doi: 10.1038/sj.jid.5700751

Figure Lengend Snippet: Figure 1. Expression of human CXCL16 in normal human skin. For chromogenic staining of CXCL16 and ADAM10 in healthy human skin, acetone-fixed sections were incubated with (a, d) rabbit anti-hCXCL16 (b) rabbit anti-ADAM10 or an irrelevant (c) rabbit IgG control antibody followed by incubation with POD-coupled goat anti-rabbit IgG antibody, enzymatic staining and counterstaining with hemalumn. In (a–c) epidermal skin and in (d) dermal skin containing a blood vessel (bottom right) is shown. Bar ¼ 50 mm.

Article Snippet: Recombinant human CXCL16 extracellular domain and chemokine domain, recombinant human IFN-g, recombinant human tumor necrosis factor-a, goat anti-human CXCL16 (hCXCL16) antibody, rat anti-murine CXCL16 (mCXCL16) mAb (clone 142417), phycoery- thrin-conjugated and unconjugated mouse anti-human CXCR6 (hCXCR6) mAb (clone 56811), and rat anti-murine CXCR6 (mCXCR6) mAb (clone 221002) were obtained from R&D Systems (Wiesbaden, Germany).

Techniques: Expressing, Staining, Incubation, Control

Journal: The Journal of investigative dermatology

Article Title: Constitutive expression and regulated release of the transmembrane chemokine CXCL16 in human and murine skin.

doi: 10.1038/sj.jid.5700751

Figure Lengend Snippet: Figure 2. In situ expression and in vivo-release of murine CXCL16. (a) Paraformaldehyde-fixed fresh frozen sections of murine skin were incubated with radioactively labeled antisense cRNA probes (as) for mCXCL16 and sense- controls (s), respectively. Hybridized cRNA probes were subsequently visualized by autoradiography. Bar ¼ 20mm. (b) Wound fluid from injured mice was collected every 24hours over a period of 15 days, cleared by centrifugation and subsequently analyzed for the presence of CXCL16 by an ELISA specific for mCXCL16. Data are given as mean and SD (n ¼ 5). Statistically significant release of mCXCL16 (Po0.05) is indicated by asterisks.

Article Snippet: Recombinant human CXCL16 extracellular domain and chemokine domain, recombinant human IFN-g, recombinant human tumor necrosis factor-a, goat anti-human CXCL16 (hCXCL16) antibody, rat anti-murine CXCL16 (mCXCL16) mAb (clone 142417), phycoery- thrin-conjugated and unconjugated mouse anti-human CXCR6 (hCXCR6) mAb (clone 56811), and rat anti-murine CXCR6 (mCXCR6) mAb (clone 221002) were obtained from R&D Systems (Wiesbaden, Germany).

Techniques: In Situ, Expressing, In Vivo, Incubation, Labeling, Autoradiography, Centrifugation, Enzyme-linked Immunosorbent Assay

Journal: The Journal of investigative dermatology

Article Title: Constitutive expression and regulated release of the transmembrane chemokine CXCL16 in human and murine skin.

doi: 10.1038/sj.jid.5700751

Figure Lengend Snippet: Figure 3. Surface expression and release of CXCL16 by cultured keratinocytes. (a) HaCaT cells were harvested and analyzed for CXCL16 surface expression by flow cytometry using a purified rabbit anti-hCXCL16 antibody that was detected by a phycoerythrin-conjugated secondary antibody. The fluorescence signal of the cells stained for hCXCL16 is shown in comparison to that of unstained cells or cells receiving an isotype control antibody. (b) HaCaT cells were incubated in serum-free medium for different periods of time in the presence or absence of marimastat (5 mM). Subsequently, conditioned media were harvested and analyzed for released CXCL16 by a specific ELISA for hCXCL16. (c) PAM212 cells were incubated with a rat anti-mCXCL16 antibody, an isotype control antibody, or left unstained. After incubation with a FITC-conjugated secondary antibody cells were analyzed by flow cytometry. (d) PAM212 cells were incubated in serum-free medium for 4 hours in the presence or absence of marimastat (5 mM). Subsequently, released CXCL16 in the conditioned media was determined by a specific ELISA for mCXCL16. Data are representative for three experiments and shown as mean and SD (n ¼ 3). Asterisks indicate statistically significant differences (Po0.05) between inhibitor-treated cells and untreated cells.

Article Snippet: Recombinant human CXCL16 extracellular domain and chemokine domain, recombinant human IFN-g, recombinant human tumor necrosis factor-a, goat anti-human CXCL16 (hCXCL16) antibody, rat anti-murine CXCL16 (mCXCL16) mAb (clone 142417), phycoery- thrin-conjugated and unconjugated mouse anti-human CXCR6 (hCXCR6) mAb (clone 56811), and rat anti-murine CXCR6 (mCXCR6) mAb (clone 221002) were obtained from R&D Systems (Wiesbaden, Germany).

Techniques: Expressing, Cell Culture, Flow Cytometry, Purification, Fluorescence, Staining, Comparison, Control, Incubation, Enzyme-linked Immunosorbent Assay

Journal: The Journal of investigative dermatology

Article Title: Constitutive expression and regulated release of the transmembrane chemokine CXCL16 in human and murine skin.

doi: 10.1038/sj.jid.5700751

Figure Lengend Snippet: Figure 4. Effect of ADAM10/ADAM17-inhibitors on CXCL16-release from cultured keratinocytes. (a) Cell lysate of cultured HaCaT cells was investigated for the presence of immature and processed forms of the metallproteinaseses ADAM10 and ADAM17. For Western blotting purified rabbit antibodies against the C-terminus of ADAM10 and ADAM17, respectively were used. (b–d) (b) HaCaT cells (c) human primary keratinocytes, (d) WT-ECV304 and CXCL16-ECV304 cells and were incubated for 4 hours in the presence or absence of 5 mM GI254023X or GW280264X. Subsequently, conditioned media were harvested and cell lysates were prepared. Released and cell-associated CXCL16 in media and lysates, respectively, was then quantified by ELISA. Statistically significant differences (Po0.05) between inhibitor-treated cells and untreated cells are indicated by asterisks.

Article Snippet: Recombinant human CXCL16 extracellular domain and chemokine domain, recombinant human IFN-g, recombinant human tumor necrosis factor-a, goat anti-human CXCL16 (hCXCL16) antibody, rat anti-murine CXCL16 (mCXCL16) mAb (clone 142417), phycoery- thrin-conjugated and unconjugated mouse anti-human CXCR6 (hCXCR6) mAb (clone 56811), and rat anti-murine CXCR6 (mCXCR6) mAb (clone 221002) were obtained from R&D Systems (Wiesbaden, Germany).

Techniques: Cell Culture, Western Blot, Purification, Incubation, Enzyme-linked Immunosorbent Assay

Journal: The Journal of investigative dermatology

Article Title: Constitutive expression and regulated release of the transmembrane chemokine CXCL16 in human and murine skin.

doi: 10.1038/sj.jid.5700751

Figure Lengend Snippet: Figure 5. Effect of ADAM10/ADAM17 downregulation by siRNA on CXCL16-release from kreatinocytes. Cultured keratinocytes prepared from human foreskin were transfected with different heteroduplexed siRNA oligonucleotide constructs for downregulation of ADAM10 (A10-1, 2, 3) or ADAM17 (A17-1, 2, 3), an irrelevant siRNA control or without siRNA. At 48 hours after transfection mRNA and protein was extracted. (a and b) The mRNA expression level of ADAM10 and ADAM17 was determined by real-time RT-PCR and expressed in relation to that of glyceraldehyde-3-phosphate dehydrogenase. (c) Lysates of siRNA-transfected cells were analyzed for expression of the pro- and mature form of ADAM10 and controlled for b-actin content by Western blotting. (d and e) For CXCL16 release experiments siRNA-treated cells were washed and incubated for 4 hours in fresh medium. (c and e) Subsequently, cells were harvested for analysis of ADAM10 and ADAM17 mRNA expression and media were collected for quantification of released CXCL16 by ELISA. Data are representative for three experiments and shown as mean and SD (n ¼ 3). Asterisks indicate statistically significant differences (Po0.05) between cells receiving specific siRNA and control cells treated with irrelevant siRNA.

Article Snippet: Recombinant human CXCL16 extracellular domain and chemokine domain, recombinant human IFN-g, recombinant human tumor necrosis factor-a, goat anti-human CXCL16 (hCXCL16) antibody, rat anti-murine CXCL16 (mCXCL16) mAb (clone 142417), phycoery- thrin-conjugated and unconjugated mouse anti-human CXCR6 (hCXCR6) mAb (clone 56811), and rat anti-murine CXCR6 (mCXCR6) mAb (clone 221002) were obtained from R&D Systems (Wiesbaden, Germany).

Techniques: Cell Culture, Transfection, Construct, Control, Expressing, Quantitative RT-PCR, Western Blot, Incubation, Enzyme-linked Immunosorbent Assay

Journal: The Journal of investigative dermatology

Article Title: Constitutive expression and regulated release of the transmembrane chemokine CXCL16 in human and murine skin.

doi: 10.1038/sj.jid.5700751

Figure Lengend Snippet: Figure 8. Activity of keratinocyte-derived CXCL16 on CXCR6-transfected HEK293 cells. CXCR6-expressing HEK293 cells were treated with recombinant CXCL16 or concentrated HaCaT-conditioned (HaCaT-CM) medium for 30 minutes at either (a) 371C or (b) 41C. In a separate experiment recombinant CXCL16 or concentrated HaCaT-conditioned medium were preincubated with a neutralizing antibody to (c) CXCL16 or an (d) isotype control and then added to the CXCR6-expressing HEK293 cells for 30 minutes at 371C. Subsequently, cells were assayed for CXCR6 surface expression using a phycoerythrin-labeled mAb to CXCR6. As a control, WT-HEK293 cells expressing no CXCR6 were stained in parallel. The mean intensity of the fluorescence signal from CXCL16 or HaCaT-CM-treated cells was calculated as percentage of that from untreated cells and is shown as insets. Data are representative for three experiments.

Article Snippet: Recombinant human CXCL16 extracellular domain and chemokine domain, recombinant human IFN-g, recombinant human tumor necrosis factor-a, goat anti-human CXCL16 (hCXCL16) antibody, rat anti-murine CXCL16 (mCXCL16) mAb (clone 142417), phycoery- thrin-conjugated and unconjugated mouse anti-human CXCR6 (hCXCR6) mAb (clone 56811), and rat anti-murine CXCR6 (mCXCR6) mAb (clone 221002) were obtained from R&D Systems (Wiesbaden, Germany).

Techniques: Activity Assay, Derivative Assay, Transfection, Expressing, Recombinant, Control, Labeling, Staining, Fluorescence

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: International journal of oncology

Article Title: Evaluation of CXCL10 as a novel serum marker for predicting liver metastasis and prognosis in colorectal cancer.

doi: 10.3892/ijo.2011.1247

Figure Lengend Snippet: Figure 2. (a), Scattergram of serum CXCL10 in stage I-IV colorectal cancer patients and control subjects (p<0.0001). (b), Scattergram of serum CXCL10 subdivided by pathological T stage (p=0.0101). (c), Scattergram of serum CXCL10 subdivided by clinical H stage (p=0.0004). (d), The relationships between serum CXCL10 and serum CEA level (r=0.2853, p<0.0001).

Article Snippet: Primary anti-CXCL10 goat polyclonal antibody (clone AF976, R&D Systems, MN, USA) and anti-CXCR3 rabbit polyclonal antibody (clone ab8023, Abcam, Cambridge, UK) at a dilution of 1:25 were used for the labeled streptavidinbiotin method (LASB2 kit/HRP, Dako Cytomation).

Techniques: Control

Journal: International journal of oncology

Article Title: Evaluation of CXCL10 as a novel serum marker for predicting liver metastasis and prognosis in colorectal cancer.

doi: 10.3892/ijo.2011.1247

Figure Lengend Snippet: Figure 3. (a), Cancer specific survival curves, for 218 patients with colorectal cancer, according to serum CXCL10 level. Patients with higher serum CXCL10 had significantly poorer prognoses than those with lower serum CXCL10 levels (p<0.0001, log-rank test); cut-off value = 199 pg/ml. (b), Cancer specific survival curves for 163 patients in stage I-III with curative intent, according to serum CXCL10 level. Patients with higher serum CXCL10 also had significantly poorer prognoses than those with lower serum CXCL10 levels (p=0.0372, log-rank test); cut-off value = 153 pg/ml.

Article Snippet: Primary anti-CXCL10 goat polyclonal antibody (clone AF976, R&D Systems, MN, USA) and anti-CXCR3 rabbit polyclonal antibody (clone ab8023, Abcam, Cambridge, UK) at a dilution of 1:25 were used for the labeled streptavidinbiotin method (LASB2 kit/HRP, Dako Cytomation).

Techniques:

Journal: International journal of oncology

Article Title: Evaluation of CXCL10 as a novel serum marker for predicting liver metastasis and prognosis in colorectal cancer.

doi: 10.3892/ijo.2011.1247

Figure Lengend Snippet: Figure 4. CXCL10 protein expression was assessed using immunohistochemistry in normal mucosa with primary CRC (a), primary colorectal cancer (b) and liver metastasis tissue (c). Magnification, x40. Scale bar = 500 µm.

Article Snippet: Primary anti-CXCL10 goat polyclonal antibody (clone AF976, R&D Systems, MN, USA) and anti-CXCR3 rabbit polyclonal antibody (clone ab8023, Abcam, Cambridge, UK) at a dilution of 1:25 were used for the labeled streptavidinbiotin method (LASB2 kit/HRP, Dako Cytomation).

Techniques: Expressing, Immunohistochemistry

Journal: Frontiers in immunology

Article Title: Metabolic Syndrome in Psoriasis Is Associated With Upregulation of CXCL16 on Monocytes and a Dysbalance in Innate Lymphoid Cells.

doi: 10.3389/fimmu.2022.916701

Figure Lengend Snippet: FIGURE 1 | Expression of CXCL16 on monocytes is associated with enhanced clinical disease activity and cardiovascular risk. (A) Flow cytometry analysis of CXCL16 expression on monocyte subpopulations defined by gating for CD14 and CD16 expression in PBMC: classical monocytes (CD14++ CD16-, R1 green), intermediate monocytes (CD14++ CD16+, R2 blue) and non-classical monocytes (CD14low CD16++, R3, red), mean and SD of healthy controls (n= 14; unfilled dots) and patients with psoriasis vulgaris (n= 39; filled dots), Welch’s t test. (B) Correlation of CXCL16 expression on all monocytes of psoriatic patients (n=39) and clinical disease activity (PASI), Pearson’s correlation. (C) Correlation for each monocyte subset defined by color indicated in (A) and PASI score, Pearson’s correlation. (D) Mean and SD of PASI (unpaired t test) and PROCAM (Mann-Whitney test) of normal-weight (unfilled triangles; n=19) and obese psoriatic patients (filled triangles; n=20). (E) Correlation of chemokine expression with cardiovascular risk (PROCAM score); and body-mass-index (BMI). Each dot represents an individual patient, Pearson correlation. (F) CXCL16 expression on monocytes in psoriatic patients without atherosclerosis (n= 6; unfilled trapezoid) or with atherosclerosis (n= 9; filled trapezoid), mean and SD, unpaired t test. *p < 0.05; **p < 0.01; ***< 0.001. ns, not significant.

Article Snippet: CD14 -APC BD Bioscience, San Jose, USA #555399 CD16 -PerCP BioLegend, San Diego, USA #302029 CXCL16 -FITC R&D Systems, Abingdon, UK #FAB976G normal goat IgG isotype -FITC R&D Systems, Abingdon, UK #Ab-108-C June 2022 | Volume 13 | Article 916701

Techniques: Expressing, Activity Assay, Flow Cytometry, MANN-WHITNEY

Journal: Frontiers in immunology

Article Title: Metabolic Syndrome in Psoriasis Is Associated With Upregulation of CXCL16 on Monocytes and a Dysbalance in Innate Lymphoid Cells.

doi: 10.3389/fimmu.2022.916701

Figure Lengend Snippet: FIGURE 2 | Phagocytosis of oxLDL by monocytes increases with CXCL16 expression. (A) PBMC were incubated with red fluorescing oxLDL for 3 h and subsequently stained for CD14, CD16 and CXCL16 expression. (B) The mean fluorescence intensity (MFI) of CXCL16 and oxLDL is shown. Every psoriasis patient is represented by three dots (n= 15). Colors represent monocyte subsets as defined in Figure 1A, Spearman correlation. (C) oxLDL concentrations in monocytes subsets of healthy controls (unfilled dots; n=5) and patients with psoriasis (filled dots; n=15), mean and SD, unpaired t test. (D) intracellular oxLDL concentrations in monocytes of normal-weight psoriatic patients (unfilled triangles; n=7) and obese patients (filled triangles; n=8), mean and SD, unpaired t test. *p < 0.05; **p < 0.01; ***p < 0.001. ns, not significant.

Article Snippet: CD14 -APC BD Bioscience, San Jose, USA #555399 CD16 -PerCP BioLegend, San Diego, USA #302029 CXCL16 -FITC R&D Systems, Abingdon, UK #FAB976G normal goat IgG isotype -FITC R&D Systems, Abingdon, UK #Ab-108-C June 2022 | Volume 13 | Article 916701

Techniques: Expressing, Incubation, Staining

Journal: Scientific reports

Article Title: Discovery of an enzyme and substrate selective inhibitor of ADAM10 using an exosite-binding glycosylated substrate.

doi: 10.1038/s41598-016-0013-4

Figure Lengend Snippet: Figure 5. Inhibition of shedding of HER2, CXCL16 and syndecan by CID 3117694. (A) Effect of CID 3117694 on viability of BT474 breast cancer cells and HEK293 cells. Please note that only 100 µM of CID 3117694 has moderate effect on HEK293 cell viability. (B) Inhibition of HER2 shedding in BT474 cells by CID 3117694. One-way analysis of variance (ANOVA) was used followed by Dunnett post hoc test. The data shown are the mean ± SEM, n = 3 from 3 independent experiments. *****p value < 0.0001, ***p value < 0.001, **p value < 0.01, *p value < 0.05; GI–control inhibitor of ADAM10 and MMPs GI254023X; (C) Results of CXCL16 ELISA assay in A549 lung cancer cells. The data shown are the mean ± SEM, n = 3 from 3 independent experiments. One-way analysis of variance (ANOVA) was used followed by Dunnett post hoc test. *****p value < 0.0001, ***p value < 0.001, **p value < 0.01, *p value < 0.05; all compared to DMSO TI. (D) Results of syndecan-4 dot blot assay (blot representative of three independent experiments shown).

Article Snippet: Serum levels of CXCL16 were measured using the DuoSet human CXCL16 Elisa Kit (R&D systems, Wiesbaden, Germany).

Techniques: Inhibition, Control, Enzyme-linked Immunosorbent Assay, Dot Blot