Journal: Physiological Reports

Article Title: CXCL10 is a novel anti‐angiogenic factor downstream of p53 in cardiomyocytes

doi: 10.14814/phy2.15304

Figure Lengend Snippet: p53‐induced CXCL10, secreted from cardiomyocytes, inhibited the tube formation of endothelial cells. Culture media were collected from p53‐ or β‐gal‐overexpressing NRCMs. RAOECs were cultured with the conditioned media in the presence of AMG487, an inhibitor of CXCR3. Angiogenesis assay was performed. (a) Representative images were shown. (b) The number of tube formation was measured ( n = 6). * p < 0.05, ** p < 0.01 by one‐way ANOVA followed by Tukey–Kramer test.

Article Snippet: After coating the wells with the extracellular matrix, 5 × 10 5 RAOECs were seeded to each well and cultured with the conditioned media contained AMG 487 (MedChemExpress), a selective antagonist of CXCR3.

Techniques: Cell Culture, Angiogenesis Assay

Journal: Journal of leukocyte biology

Article Title: Hyperforin down-regulates effector function of activated T lymphocytes and shows efficacy against Th1-triggered CNS inflammatory-demyelinating disease.

doi: 10.1189/jlb.0707469

Figure Lengend Snippet: Fig. 2. Hyp-DCHA down-regulates CXCR3 expression on activated T cells. Surface expression of CXCR3 on the membrane of 72 h PHA and IL-2- activated T cells as detected by FACS is shown. CXCR3 expression was evaluated on cells treated 24 h with 2.5 M Hyp-DCHA (upper panel); the gray peak shows the fluorescence of cells incubated with FITC-conjugated isotype control mAb (anti-IgG1). Unimodal distribution with a statistically significant shift (down-regulation) between a Hyp-DCHA-treated and untreated sample is observed. The down-regulation of CCR3 after 24 h of treatment is not significant (lower panel). Representative results of four independent experi- ments are shown.

Article Snippet: The cells were treated with commercially available FITC- or PE-conjugated mAb: anti-CD3, -CD4, and -CD8, isotype-matched controls (BD PharMingen, San Diego, CA, USA), anti-IL-4 and -IFN- (BD PharMingen), FITC-conjugated anti-human CXCR3 (clone 49801.111), and anti-human CCR3 (R&D Systems Inc., Minneapolis, MN, USA).

Techniques: Expressing, Membrane, Incubation, Control

Journal: Clinical Cancer Research

Article Title: Role of CXCR3 Ligands in IL-7/IL-7Rα-Fc–Mediated Antitumor Activity in Lung Cancer

doi: 10.1158/1078-0432.ccr-10-3346

Figure Lengend Snippet: Figure 4. CXCR3 pathway is required for IL-7/IL-7Ra-Fc–mediated antitumor activity. For in vivo neutralizations, mice bearing 5-day established subcutaneous tumors were treated with IL-7/IL7Ra-Fc on days 5 and 10. Twenty four hours prior to IL-7/IL7Ra-Fc treatment, and then 3 times per week, mice were injected i.p. individually with the respective specific or appropriate control antibodies for the duration of the experiment. A and B, neutralization of CXCL9 (A) or CXCL10 (B) or IFNg (B) reversed the antitumor benefit of IL-7/IL-7Ra-Fc [bottom, photograph of tumors from diluents, IL-7/IL-7Ra-Fc, and IL-7/IL-7Ra-Fc þ anti-CXCL9, or anti-CXCL10 or anti-IFNg (IL-7/IL-7Ra-Fc þ control Ab was the same as IL-7/IL-7Ra-Fc treated group, data not shown)]. C, CXCL9, CXCL10, and IFNg were reduced in tumors (day 15) following treatment of the tumor bearing mice with IL-7/IL-7Ra-Fc and the respective neutralizing antibodies: anti-CXCL9, or anti-CXCL10 or anti-IFNg antibody in comparison to control Abs. D, neutralization of CXCL9, CXCL10, or IFNg reduced the frequency of CXCR3 activated CD8 T cells in the tumor (day 15). Data; mean SEM; *, P < 0.01 for IL-7/IL-7Ra-Fc þ control Ab compared with IL-7/IL- 7Ra-Fc þ anti-cytokine Ab or Diluent groups (n ¼ 6/group).

Article Snippet: Anti-mouse mAbs to CXCR3 (220803), IL-7Ra-Fc chimeric molecule (747-MR; endotoxin level: less than 1.0 EU/mg of protein by LAL method), ELISA antibody pairs for murine IFNg , CXCL9, CXCL10, and IL12 were purchased from R&D Systems, sensitivity of 3 to 5 pg/mL.

Techniques: Activity Assay, In Vivo, Injection, Control, Neutralization, Comparison

Journal: The Korean journal of internal medicine

Article Title: Therapeutic effect of multiple functional minicircle vector encoding anti-CD25/IL-10/CXCR3 in allograft rejection model.

doi: 10.3904/kjim.2021.299

Figure Lengend Snippet: Figure 1. Diagram of the novel multiple target-directed agent (MTA), which was constructed as an anti-CD25 antibody conju- gated to interleukin-10 (IL-10) and C-X-C motif chemokine recep- tor 3 (CXCR3) in turn at the end of the Fab region. MTA structure designed by the fusion of the anti-CD25 antibody and the dimer form of IL-10 via short flexible linkers, called (G4S1)3 linker. CXCR3 was connected with IL-10 using the (G4S1)2 linker. (G4S1)3, (GG- GGS)3; (G4S1)2, (GGGGS)2; mIL-10, mouse intrleukin-10; mCXCR3, mouse C-X-C motif chemokine receptor 3; ECD1, extracellular domain 1.

Article Snippet: Briefly, a 96-well microtiter plate was coated with each commercial anti-mouse IL-10 (MAB417-100, R&D Systems, Minneapolis, MN, USA; at a concentration of 1 mg/mL) or anti-mouse CXCR3 (NBP2-41250, Novus Biologicals, Centennial, CO, USA; at a concentration of 1 mg/mL) in coating buffer and was incubated overnight at 4°C.

Techniques: Construct

Journal: The Korean journal of internal medicine

Article Title: Therapeutic effect of multiple functional minicircle vector encoding anti-CD25/IL-10/CXCR3 in allograft rejection model.

doi: 10.3904/kjim.2021.299

Figure Lengend Snippet: Figure 3. Detection of multiple target-directed agent (MTA) derived from minicircles (MCs) in vitro. (A) Fluorescence images of the ex- pression of red fluorescence protein (RFP). HEK293T cells were transfected with the indicated DNA, and RFP expression was analyzed using fluorescence microscopy at 3 days post-transfection. HEK293T cells that were not transfected with DNA were used as a nil group. Scale bar = 50 μm. (B, C, D) Concentrations of anti-CD25, interleukin-10 (IL-10), and C-X-C motif chemokine receptor 3 (CXCR3) in con- ditioned media of cells transfected with MC-mock or MC-MTA. Protein secretion from HEK293T cells was assessed using enzyme-linked immunosorbent assay (ELISA) at 3 days after transfection. Values are mean ± standard error. All results are representative of at least three independent experiments. DIC, differential interference contrast; PP, parent plasmid. ap < 0.05 vs. MC-mock.

Article Snippet: Briefly, a 96-well microtiter plate was coated with each commercial anti-mouse IL-10 (MAB417-100, R&D Systems, Minneapolis, MN, USA; at a concentration of 1 mg/mL) or anti-mouse CXCR3 (NBP2-41250, Novus Biologicals, Centennial, CO, USA; at a concentration of 1 mg/mL) in coating buffer and was incubated overnight at 4°C.

Techniques: Derivative Assay, In Vitro, Fluorescence, Transfection, Expressing, Microscopy, Enzyme-linked Immunosorbent Assay, Plasmid Preparation

Journal: iScience

Article Title: Naive and memory CD4 + T cell subsets can contribute to the generation of human Tfh cells

doi: 10.1016/j.isci.2021.103566

Figure Lengend Snippet: CyTOF antibody panel

Article Snippet: Vio Bright FITC anti-human CXCR3 , Miltenyi Biotec , Cat# 130-118-673; RRID:AB_2734057.

Techniques:

Journal: iScience

Article Title: Naive and memory CD4 + T cell subsets can contribute to the generation of human Tfh cells

doi: 10.1016/j.isci.2021.103566

Figure Lengend Snippet: Selected genes involved in Tfh cell biology

Article Snippet: Vio Bright FITC anti-human CXCR3 , Miltenyi Biotec , Cat# 130-118-673; RRID:AB_2734057.

Techniques: Activation Assay

Journal: iScience

Article Title: Naive and memory CD4 + T cell subsets can contribute to the generation of human Tfh cells

doi: 10.1016/j.isci.2021.103566

Figure Lengend Snippet: Distinct CD4 + T cell subsets contribute to the generation of Tfh with heterogeneous functional profiles (A) Mean fluorescence intensity of the CXCR5 marker expressed by CXCR5 + PD-1 + cells derived from (1) naive, (2) MemPD-1 neg , (3) MemPD-1 neg and Tfh. (B) Frequency of IL-21- and/or IFNγ-positive cells among CXCR5 + PD-1 + cells at day 3. (C – E) Representative flow plots showing CXCR3, ICOS, and CD40L expression by CXCR5 + PD-1 + cells (left panel) and frequency of CXCR3-, ICOS-, and CD40L-positive cells among CXCR5 + PD-1 + cells at day 3 (right panel). (F) Ex vivo cells or their respective Tfh D3 counterparts obtained after 3 days of splenocyte culture were co-cultured with autologous B cells for 7 days. (G) Box plots represent the frequency of CD27 + CD38 + cells among CD19 + cells, the concentration of total immunoglobulins and the absolute number of live B cells after co-culture. (H) Quantification of IgG1, IgG4, and IgA in the co-culture supernatants. Each symbol (A–H) represents an individual donor. (A–H) A Wilcoxon matched pairs test was performed, ∗, p < 0.05; ∗∗, p < 0.005; ∗∗∗, p < 0.001.

Article Snippet: Vio Bright FITC anti-human CXCR3 , Miltenyi Biotec , Cat# 130-118-673; RRID:AB_2734057.

Techniques: Functional Assay, Fluorescence, Marker, Derivative Assay, Expressing, Ex Vivo, Cell Culture, Concentration Assay, Co-Culture Assay

Journal: iScience

Article Title: Naive and memory CD4 + T cell subsets can contribute to the generation of human Tfh cells

doi: 10.1016/j.isci.2021.103566

Figure Lengend Snippet:

Article Snippet: Vio Bright FITC anti-human CXCR3 , Miltenyi Biotec , Cat# 130-118-673; RRID:AB_2734057.

Techniques: Cell Analysis, Purification, Virus, Recombinant, Blocking Assay, Antibody Labeling, Transfection, Software

Journal: Journal for immunotherapy of cancer

Article Title: Systemic and intravesical adoptive cell therapy of tumor-reactive T cells can decrease bladder tumor growth in vivo.

doi: 10.1136/jitc-2020-001673

Figure Lengend Snippet: Figure 5 CXCR3 mediates T cell infiltration into orthotopic ovalbumin expressing MB49 tumor cell line (MB49OVA) bladder tumor from intravesical space. (A) RNA was isolated from MB49 and MB49OVA tumor cells and analyzed for expression of chemokines CXCL9, CXCL10 and CXCL11 as determined by PCR. (B) OT-I T cells were isolated from spleen of transgenic mice and tumor-infiltrating lymphocytes (TILs) were isolated from MB49 orthotopic tumors. Expression of CXCR3 was determined on these cell populations by flow cytometry. (C) TIL samples expanded from patient bladder tumors were analyzed for CXCR3 expression by flow cytometry. In vivo, mice were treated with OT-I T cells coated with a CXCR3 blocking antibody or rat IgG (rIgG). (D) Tumors were collected 3 and 24 hours after infusion and analyzed for CD45.1+ T cell infiltration by flow cytometry. (E) Tumor volume was measured by ultrasound. Precoating OT-I T cells with αCXCR3 decreased T cell infiltration into the tumor and abrogated the decrease in tumor growth seen with OT-I T cells coated with a control rIgG antibody. N=5 per group. Repeated 2 times. ns, not significant.

Article Snippet: OT- I T cells were isolated as described and incubated for 30 min on ice with either normal rat IgG isotype control or CXCR3 blocking antibody (500 μg, catalog# BE0249, BioXcell).

Techniques: Expressing, Isolation, Transgenic Assay, Flow Cytometry, In Vivo, Blocking Assay, Control

Journal: BMC Cancer

Article Title: Intracellular coexpression of CXC- and CC– chemokine receptors and their ligands in human melanoma cell lines and dynamic variations after xenotransplantation

doi: 10.1186/1471-2407-14-118

Figure Lengend Snippet: Surface expression of chemokine receptors

Article Snippet: Mouse monoclonal antibodies (mAb) against human CXCR3, CXCR4, CXCR7, CCR7, CXCL12, CCL19, CCL27 were from R&D Systems (Minneapolis, MN, USA) and mouse mAb against human CXCL10 was from BD Biosciences (Franklin Lakes, NJ, USA).

Techniques: Expressing

Journal: BMC Cancer

Article Title: Intracellular coexpression of CXC- and CC– chemokine receptors and their ligands in human melanoma cell lines and dynamic variations after xenotransplantation

doi: 10.1186/1471-2407-14-118

Figure Lengend Snippet: Surface expression of chemokine receptors. Representative examples for the quantification of chemokine receptors surface expression by flow cytometry are shown. Overlaid histograms of PE fluorescence of specific anti-receptor monoclonal antibody (continuous red line) and correspondent isotypic control (discontinuous black line) show the control Hut-78 cell line with a high percentage of CXCR3+ cells (A) , WM-115 with no expression of CXCR4 (B) and IGR-39 with a small subpopulation of CXCR7+ cells (C) , which is further illustrated in a biparametric dotplot of FS vs PE fluorescence (D) .

Article Snippet: Mouse monoclonal antibodies (mAb) against human CXCR3, CXCR4, CXCR7, CCR7, CXCL12, CCL19, CCL27 were from R&D Systems (Minneapolis, MN, USA) and mouse mAb against human CXCL10 was from BD Biosciences (Franklin Lakes, NJ, USA).

Techniques: Expressing, Flow Cytometry, Fluorescence, Control

Journal: BMC Cancer

Article Title: Intracellular coexpression of CXC- and CC– chemokine receptors and their ligands in human melanoma cell lines and dynamic variations after xenotransplantation

doi: 10.1186/1471-2407-14-118

Figure Lengend Snippet: Intracellular expression of chemokine receptors

Article Snippet: Mouse monoclonal antibodies (mAb) against human CXCR3, CXCR4, CXCR7, CCR7, CXCL12, CCL19, CCL27 were from R&D Systems (Minneapolis, MN, USA) and mouse mAb against human CXCL10 was from BD Biosciences (Franklin Lakes, NJ, USA).

Techniques: Expressing