cxcl13 level  (OriGene)

Journal: Theranostics

Article Title: Kupffer cells promote T-cell hepatitis by producing CXCL10 and limiting liver sinusoidal endothelial cell permeability

doi: 10.7150/thno.44960

Figure Lengend Snippet: Dynamic changes of immune cell infiltration, and cytokine & chemokine profile in the liver after the KC depletion in steady condition. (A) Numbers of infiltrating immune cells (monocytes, T cells, B cells, NKT cells and NK cells) in livers before ILY injection (0 h) or at 14 and 24 h after 3 ILY injections (100 ng/g, i.p. at 2 h intervals) to ihCD59 + and LysM-Cre + ihCD59 + mice (n=4 at 0 h and n=6 for all other time points). (B-D) RT2 Profiler PCR Array analysis of the hepatic cytokine and chemokine transcripts in the LysM-Cre + ihCD59 + and ihCD59 + mice at 2 h after 1 ILY injection (B, 100 ng/g, i.p., n = 2), and at 14 h (C) and 24 h (D) after 3 ILY injections (100 ng/g, i.p. at 2 h intervals, n = 2 for each time point). Scatter plot represents the relative transcript levels for each gene obtained from LysM-Cre + ihCD59 + mice plotted against the same gene from ihCD59 + mice. Genes with fold changes >2.0 are shown in the table on the right. (E) ELISA analysis of CXCL13 in serum collected from ihCD59 + and LysM-Cre + ihCD59 + mice (n = 3) at 8 h after 3 ILY injections (100 ng/g, i.p. at 2 h intervals). Data are shown as mean ± SEM. * P<0.05; ** P<0.01, as determined by unpaired two-tailed Student's t-test.

Article Snippet: In some experiments, serum CXCL13 level was measured using a mouse CXCL13/BLC/BCA-1 DuoSet Elisa Kit (R&D DY470, Minneapolis, MN).

Techniques: Injection, Enzyme-linked Immunosorbent Assay, Two Tailed Test

Journal: Medicine

Article Title: Study on correlation between CXCL13 and prognosis and immune characteristics of ovarian cancer

doi: 10.1097/MD.0000000000040272

Figure Lengend Snippet: (A) Protein–protein intersection (PPI) network of differentially expressed genes (DEGs); (B) twenty core genes in the PPI network; (C) prognostic DEGs; (D) core genes CXCL13 and CD3E on the Venn diagram; (E) meta-analysis for OS based on expression levels of CXCL13 in OC patients.

Article Snippet: The Human Protein Atlas (HPA) ( http://proteinatlas.org ) [ ] was used to analyze CXCL13 protein levels.

Techniques: Expressing

Journal: Medicine

Article Title: Study on correlation between CXCL13 and prognosis and immune characteristics of ovarian cancer

doi: 10.1097/MD.0000000000040272

Figure Lengend Snippet: TCGA and GETx datasets show expression of CXCL13 is statistical different between OC and normal tissue (A) and in different pathological stages of OC (B). (C) Oncomine database verifies the expression of CXCL13 ; (D) the negative staining of CXCL13 in 2 normal tissues; (E) the negative (0), low (1), and medium (2) immunohistochemical staining of CXCL13 in 3 OC patients.

Article Snippet: The Human Protein Atlas (HPA) ( http://proteinatlas.org ) [ ] was used to analyze CXCL13 protein levels.

Techniques: Expressing, Negative Staining, Immunohistochemical staining, Staining

Journal: Medicine

Article Title: Study on correlation between CXCL13 and prognosis and immune characteristics of ovarian cancer

doi: 10.1097/MD.0000000000040272

Figure Lengend Snippet: Survival curves of OC patients in CXCL13 high- and low-expression groups.

Article Snippet: The Human Protein Atlas (HPA) ( http://proteinatlas.org ) [ ] was used to analyze CXCL13 protein levels.

Techniques: Expressing

Journal: Medicine

Article Title: Study on correlation between CXCL13 and prognosis and immune characteristics of ovarian cancer

doi: 10.1097/MD.0000000000040272

Figure Lengend Snippet: Gene set enrichment analysis enrichment pathway of CXCL13 expression profile in OC patients.

Article Snippet: The Human Protein Atlas (HPA) ( http://proteinatlas.org ) [ ] was used to analyze CXCL13 protein levels.

Techniques: Expressing

Journal: Medicine

Article Title: Study on correlation between CXCL13 and prognosis and immune characteristics of ovarian cancer

doi: 10.1097/MD.0000000000040272

Figure Lengend Snippet: Correlation between the expression of CXCL13 and infiltration of various immune cell components in the tumor microenvironment. (A) CD8 + T cell; (B) activated CD4 + memory T cell; (C) memory B cell; (D) M1 macrophage cell; (E) follicular helper T cell; (F) resting CD4 + memory T cell; (G) naive B cell; (H) M2 macrophage cell; (I) activated myeloid dendritic cell; (J) neutrophils.

Article Snippet: The Human Protein Atlas (HPA) ( http://proteinatlas.org ) [ ] was used to analyze CXCL13 protein levels.

Techniques: Expressing

Journal: Medicine

Article Title: Study on correlation between CXCL13 and prognosis and immune characteristics of ovarian cancer

doi: 10.1097/MD.0000000000040272

Figure Lengend Snippet: (A) Correlation between the expression of CXCL13 and PD-1 of TCGA-OC samples; (B) correlation between the expression of CXCL13 and CR/PR, SD/PD in mUC samples after receiving PD-L1 blocking therapy (CR = complete response, PR = partial response, PD = progressive disease, SD = stable disease). Survival curves of mUC patients with high- and low- CXCL13 expression in SD/PD (C) and CR/PR (D) group.

Article Snippet: The Human Protein Atlas (HPA) ( http://proteinatlas.org ) [ ] was used to analyze CXCL13 protein levels.

Techniques: Expressing, Blocking Assay

Journal: Frontiers in Immunology

Article Title: Role of CXCL5 in Regulating Chemotaxis of Innate and Adaptive Leukocytes in Infected Lungs Upon Pulmonary Influenza Infection

doi: 10.3389/fimmu.2021.785457

Figure Lengend Snippet: CXCL5 regulated CXCL13 expression in pulmonary CD64+ macrophages/monocytes via the CXCL1/2/5-CXCR2 signaling pathway. (A–C) The protein levels of CXCL1/2 (A) , CXCL12 (B) , and CXCL13 (C) in the BALF of infected mice were measured by ELISA, and the level of CXCL5 was used as a reference (n=5). (D) RT-qPCR for IFN-α and IFN-β mRNA was performed in lung homogenates from WT and CXCL5 -/- mice before and after (3 d.p.i.) influenza infection. The fold changes in the mRNA expression levels of these genes were calculated using the 2 -ΔΔCt method of relative quantification with GAPDH as the endogenous reference gene. The relative fold changes in IFN-α and IFN-β expression in infected mice compared to normal mice are presented (n=4). (E, F) Percentages and numbers of CXCL13-expressing cells among single lung cells from WT and CXCL5 -/- mice at 3 d.p.i. (E) and 8 d.p.i. (F) as assessed by intracellular staining of CXCL13 protein (APC-CXCL13) and flow cytometry (n=4). (G, H) Percentages and numbers of CXCL13-expressing cells among pulmonary CD64+ cells from WT and CXCL5 -/- mice at 3 d.p.i. (G) and 8 d.p.i. (H) as assessed by intracellular staining of CXCL13 protein (APC-CXCL13) and flow cytometry (n=4). (I) Percentages and numbers of CXCL13-expressing cells among pulmonary CD11c+ cells from WT and CXCL5 -/- mice at 8 d.p.i. as assessed by intracellular staining of CXCL13 protein (APC-CXCL13) and flow cytometry (n=4). (J–L) Percentages and numbers of CD206 (J) , CD44 (K) , and CD274 (L) surface markers among pulmonary CD64+ cells from WT and CXCL5-/- mice at 3 d.p.i. (n=4). (M) Detection of the expression of mouse CXCR2 on total AMs and CD64+ AMs by western blotting. (N) RT-qPCR was performed on cell lysates of cultured CD64+ macrophages after 10 h of influenza challenge or unstimulated (control) conditions, and the fold changes were calculated as described in the MATERIALS AND METHODS section (n=4). (O) Expression levels of the indicated chemokines from supernatants of cultured CD64+ macrophages after 20 h of influenza challenge or unstimulated (control) conditions as determined by ELISA (n=4). (P) ELISA was used to determine the expression levels of CXCL13 protein in supernatants of cultured CD64+ macrophages after 20 h of influenza challenge and treatment with the indicated recombinant chemokine proteins, the CXCR2 antagonist SB225002, the PI3K inhibitor LY294002, and the MEK inhibitor PD98059 (n=3). The error bars represent the SDs. * P < 0.05 based on Student’s t -test.

Article Snippet: CXCL5 (MX000), CXCL1 (MKC00B), CXCL2 (MM200), CXCL12 (MCX120), and CXCL13 (MCX130) levels were determined using ELISA kits from R&D Systems (MN, USA) in accordance with the manufacturer’s instructions.

Techniques: Expressing, Infection, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Quantitative Proteomics, Staining, Flow Cytometry, Western Blot, Cell Culture, Control, Recombinant

Journal: Frontiers in Immunology

Article Title: Role of CXCL5 in Regulating Chemotaxis of Innate and Adaptive Leukocytes in Infected Lungs Upon Pulmonary Influenza Infection

doi: 10.3389/fimmu.2021.785457

Figure Lengend Snippet: CXCL5-CXCR2 signaling regulated pulmonary neutrophil and B cell recruitment and CXCL13 production from lung macrophages upon H1N1 infection. (A–E) WT mice were infected with H1N1 virus (1000 CCID 50 ) and treated with either CXCR2 antagonist or PBS as a control during 1-3 d.p.i. Microscopic view of BALF leukocytes from the infected mice collected at 3 d.p.i. and cytospun (×400 magnification) (A) . The protein level of CXCL13 in the BALF of the infected mice was measured at 3 d.p.i. by ELISA (n=4) (B) . Percentages and numbers of CXCL13-expressing cells among pulmonary CD64+ cells (C) , CD19+ B cells (D) , and CD44+/CD274+ CD64+ cells (E) from the infected mice at 3 d.p.i. by flow cytometry and counting (n=4). (F–J) WT mice were infected with H1N1 virus (1000 CCID 50 ) and orotracheally instilled with either CXCL5 recombinant protein or PBS as a control during 5-8 d.p.i. Microscopic view of BALF leukocytes from the infected mice collected at 8 d.p.i. and cytospun (×400 magnification) (F) . The protein level of CXCL13 in the BALF of the infected mice was measured at 8 d.p.i. by ELISA (n=4) (G) . Percentages and numbers of CD19+ B cells from the infected mice at 8 d.p.i. by flow cytometry and counting (n=4) (H) . Measurements of H1N1-specific IgG (I) and IgA (J) in the BALF of the CXCL5 treatment and control mice at 8 d.p.i. (n = 4). The error bars represent the SDs of 4 samples. * P < 0.05 based on Student’s t -test.

Article Snippet: CXCL5 (MX000), CXCL1 (MKC00B), CXCL2 (MM200), CXCL12 (MCX120), and CXCL13 (MCX130) levels were determined using ELISA kits from R&D Systems (MN, USA) in accordance with the manufacturer’s instructions.

Techniques: Infection, Virus, Control, Enzyme-linked Immunosorbent Assay, Expressing, Flow Cytometry, Recombinant

Journal: Molecular Therapy Oncolytics

Article Title: CXCR5 guides migration and tumor eradication of anti-EGFR chimeric antigen receptor T cells

doi: 10.1016/j.omto.2021.07.003

Figure Lengend Snippet: Restricted normal T cell expression of CXCR5 and upregulation of CXCL13 in non-small cell lung cancer (NSCLC) (A) The expression of CXCL13 in patients with lung adenocarcinoma (LUAD) and lung squamous cell carcinoma (LUSC) using the online tool of GEPIA. (B) CXCL13 protein expressions in NSCLC tissues were confirmed by immunohistochemistry on two tissue microarray slides (NSC157 and LC20813b). The intensity of immunostaining was graded as follows: −, negative; +, weak; ++, moderate; or +++, strong. (C) Expression of CXCL13 by immunohistochemistry. The subpanels show negative expression of CXCL13 (−), weak (+), moderate (++), and strong (+++) expressions of CXCL13 in tumor tissues ( ×400). (D) ELISA quantification of the level of CXCL13 protein in plasma samples (healthy donors n = 34, NSCLC patient donors n = 95). Single dot represents individual plasma sample. Error bars represent mean ± SD. ∗∗∗p < 0.001. (E) FACS analysis of the expression of different chemokine receptors from resting and activated T cells. Single dot represents individual sample. Error bars represent mean ± SD for each T cell population (n = 12).

Article Snippet: The CXCL13 level in the blood plasma was quantified by the CXCL13 ELISA kit from OriGene (catalog number [cat. no.] EA800062) according to the manufacturer’s instructions.

Techniques: Expressing, Immunohistochemistry, Microarray, Immunostaining, Enzyme-linked Immunosorbent Assay

Journal: Molecular Therapy Oncolytics

Article Title: CXCR5 guides migration and tumor eradication of anti-EGFR chimeric antigen receptor T cells

doi: 10.1016/j.omto.2021.07.003

Figure Lengend Snippet: Evaluation of the antitumor efficacy and chemotaxis migration of EGFR-CXCR5-CAR-T cells in vitro (A) Analysis of the cytotoxicity of EGFR-CXCR5-CAR-T cells against PC9, A549, and K562 cells. Tumor cell killing was measured via an IncuCyte assay over 48 h. SYTOX Green and CellTrace Far Red double-positive tumor cells (yellow) were calculated. Error bars represent mean ± SD for each time point. (B) Real-time cell killing image. Target cells were red, and CAR-T cells were green. (C) Cytokine production by CAR-T cells co-cultured with PC9 tumor cells. CAR-T cells were co-cultured 10:1 with tumor cells in 96-well plates for 20 h. Levels of IFN-γ and IL-2 in supernatant were determined by ELISA. Error bars represent mean ± SD for each group. (D) Chemotaxis migration of CAR-Ts toward various concentrations of recombinant human CXCL13 at different time courses of 4 h, 8 h, and 16 h. Error bars represent mean ± SD for each group (n = 3). ∗p < 0.05 derived via unpaired t test. (E) CAR-T cell proliferation assay with indicated CAR-T cells cocultured with various concentrations of recombinant human CXCL13.

Article Snippet: The CXCL13 level in the blood plasma was quantified by the CXCL13 ELISA kit from OriGene (catalog number [cat. no.] EA800062) according to the manufacturer’s instructions.

Techniques: Chemotaxis Assay, Migration, In Vitro, Cell Culture, Enzyme-linked Immunosorbent Assay, Recombinant, Derivative Assay, Proliferation Assay

Journal: Molecular Therapy Oncolytics

Article Title: CXCR5 guides migration and tumor eradication of anti-EGFR chimeric antigen receptor T cells

doi: 10.1016/j.omto.2021.07.003

Figure Lengend Snippet: In vivo tracking of the migration of 89 Zr-oxine-labeled CAR-T to A549 and A549-CXCL13 tumors using micro-PET/CT scan (A) EGFR expression in the A549 cell line stably expressing the CXCL13 gene (A549-CXCL13) after lentiviral transduction and selection. (B) Increased secretion of CXCL13 generated by A549-CXCL13 cells. ∗∗∗p < 0.001. (C) The effects of 89 Zr-oxine labeling on T cell proliferation. (D) Whole-body PET imaging, quantitative PET analysis, and biodistribution of 89 Zr-labeled T cells in tumor-bearing mice. 89 Zr-labeled mock T cells, 89 Zr-EGFR-CAR-T cells, or 89 Zr-EGFR-CXCR5-CAR-T cells were tail-vein injected into NSG mice inoculated with A549 tumor cells at the left and A549-CXCL13 tumor cells at the right side. Isotopic distribution of 89 Zr was quantified and plotted in a coronal horizon map at different time points of 2, 24, 72, and 168 h post-injection. The red and green circles represent the A549 tumor region and A549-CXCL13 tumor region, respectively. (E) Accumulated isotope signaling in A549 tumor region (green circle) and A549-CXCL13 tumor region (red circle). The percentage injection dose rate ([%ID]/g value) was calculated. Error bars represent mean ± SD for each group (n = 3).

Article Snippet: The CXCL13 level in the blood plasma was quantified by the CXCL13 ELISA kit from OriGene (catalog number [cat. no.] EA800062) according to the manufacturer’s instructions.

Techniques: In Vivo, Migration, Labeling, Micro-PET, Computed Tomography, Expressing, Stable Transfection, Transduction, Selection, Generated, Imaging, Injection

Journal: Molecular Therapy Oncolytics

Article Title: CXCR5 guides migration and tumor eradication of anti-EGFR chimeric antigen receptor T cells

doi: 10.1016/j.omto.2021.07.003

Figure Lengend Snippet: Antitumor efficacy of CAR-T cells in vivo (A) Serial bioluminescence imaging of NSG mice injected subcutaneously with A549 luc cells on the left flank and A549 luc -CXCL13 cells on the right flank. 10 days after tumor engraftment, the mice were injected with 5 × 10 5 CAR + T cells as indicated. n = 5 mice per group. Error bars represent mean ± SD for each time point (n = 5). (B) The tumor volume of the left tumors (A549 luc ) and right tumors (A549 luc -CXCL13) over 28 days was measured. Error bars represent mean ± SD for each time point (n = 5). (C) The copy number of CAR gene in the left and right tumor tissues was analyzed. ∗∗p < 0.01.

Article Snippet: The CXCL13 level in the blood plasma was quantified by the CXCL13 ELISA kit from OriGene (catalog number [cat. no.] EA800062) according to the manufacturer’s instructions.

Techniques: In Vivo, Imaging, Injection

Journal: Molecular Therapy Oncolytics

Article Title: CXCR5 guides migration and tumor eradication of anti-EGFR chimeric antigen receptor T cells

doi: 10.1016/j.omto.2021.07.003

Figure Lengend Snippet: Addition of CXCR5 facilitates T cell migration The chemokine CXCL13 is highly expressed in various tumors including lung carcinoma, whereas the classical CAR-T does not effectively infiltrate into tumor regions due to the absence of CXCR5 receptor expression. Chemotactic movement is a taxis in response to a chemical concentration gradient. When CAR-T cells are modified with the CXCR5 receptor, the motorized CAR-T cells could infiltrate into the tumor site along the gradient of CXCL13 to further clear the tumor cells.

Article Snippet: The CXCL13 level in the blood plasma was quantified by the CXCL13 ELISA kit from OriGene (catalog number [cat. no.] EA800062) according to the manufacturer’s instructions.

Techniques: Migration, Expressing, Concentration Assay, Modification

Journal: Frontiers in Immunology

Article Title: Ischemia Reperfusion Injury Triggers CXCL13 Release and B-Cell Recruitment After Allogenic Kidney Transplantation

doi: 10.3389/fimmu.2020.01204

Figure Lengend Snippet: Cytokine levels in serum samples and kidney tissue. Serum levels of pro-inflammatory cytokines were measured in models of uni- and bilateral renal IRI. CXCL13 levels significantly increased within 24 h in 30 min bilateral IRI as compared to 15 min IRI or baseline (BL) levels prior to IRI (A) . In unilateral IRI for 35 and 45 min a time-dependent increase of serum CXCL13 levels were observed 24 h after IRI (B) . To longitudinally determine the kinetics of CXCL13 release in 45 min unilateral IRI blood samples were taken at the indicated times. A maximum was measured 24 h after IRI (C) . MCP-1 (D) and IL-6 (E) were measured in comparison to sham surgery prior to IRI and 24 h after bilateral IRI. Both markers were significantly increased in comparison to baseline. 24 h after unilateral IRI a significant increase of CXCL13 (F) and of IL-6 (G) mRNA expression in renal tissue was observed in the 45 min unilateral IRI model ( n = 6–10 mice per group, one-way ANOVA, * p < 0.05; ** p < 0.01, *** p < 0.001). BL, baseline.

Article Snippet: CXCL13 serum levels were analyzed by ELISA (R&D Systems, MCX130) as described previously ( ).

Techniques: Comparison, Expressing

Journal: Frontiers in Immunology

Article Title: Ischemia Reperfusion Injury Triggers CXCL13 Release and B-Cell Recruitment After Allogenic Kidney Transplantation

doi: 10.3389/fimmu.2020.01204

Figure Lengend Snippet: Serum CXCL13 levels in mouse ktx. Post ktx levels of serum CXCL13 at day 1 were significantly increased compared to baseline. A higher increase was observed in longer cold ischemia time (30 vs. 60 min cold ischemia time). Isogenic ktx with prolonged cold ischemia time of 60 min had significantly lower CXCL13 levels compared to allogenic ktx (A) . PAS stain at day 7 revealed enhanced cell infiltration in allogenic compared to isogenic ktx (B) . Double staining for CD3+ T-lymphocytes (green) and CD45R+ B-cells (red) was performed at day 7. More interstitial CD3+ T-lymphocytes were observed in allografts compared to isografts. Allografts exhibited scattered B-cells in interstitial tissue, but also clusters of CD45R+ cells. Isografts showed only few B cells in the interstitium at day 7 (B) (bar: 100 μm, n = 6 per group, one-way ANOVA * p < 0.05; ** p < 0.01; *** p < 0.001). BL, baseline.

Article Snippet: CXCL13 serum levels were analyzed by ELISA (R&D Systems, MCX130) as described previously ( ).

Techniques: Staining, Double Staining