goat polyclonal anti cxcl13 antibody  (R&D Systems)

Journal: Frontiers in Immunology

Article Title: CXCL13 as a simple and promising blood biomarker for differentiating Sézary syndrome from mycosis fungoides and other confounding chronic inflammatory skin diseases

doi: 10.3389/fimmu.2026.1804103

Figure Lengend Snippet: CXCL13 mRNA expression in PBMCs from patients with Sézary syndrome and other confounding skin diseases. (A) Box-and-whisker plots showing individual CXCL13 ΔCt values in PBMCs from patients with Sézary syndrome (SS), mycosis fungoides (MF), atopic dermatitis (AD), psoriasis (PS) and healthy donors (HD). ΔCt values were calculated using GAPDH as housekeeping gene. Statistical significance was assessed using the Kruskal–Wallis test followed by Dunn’s multiple comparisons test. ***p ≤ 0.001; *p ≤ 0.05. (B) Relative CXCL13 expression levels expressed as fold change (RQ) for each disease group relative to HD, calculated using the mean ΔCt value of HD as reference. RQmin and RQmax represent the minimum and maximum relative quantities obtained by adding or subtracting, respectively, the standard deviation of ΔCt values to ΔΔCt.

Article Snippet: Sections were then incubated overnight at 4 °C with a goat polyclonal anti-CXCL13 antibody (AF801; R&D Systems) at a concentration of 2.5 μg/mL in 2% bovine serum albumin (BSA).

Techniques: Expressing, Whisker Assay, Standard Deviation

Journal: Frontiers in Immunology

Article Title: CXCL13 as a simple and promising blood biomarker for differentiating Sézary syndrome from mycosis fungoides and other confounding chronic inflammatory skin diseases

doi: 10.3389/fimmu.2026.1804103

Figure Lengend Snippet: CXCL13 immunohistochemical expression in skin biopsies from Sézary syndrome, mycosis fungoides and inflammatory skin diseases. Representative immunohistochemical staining for CXCL13 in skin lesions from patients with Sézary syndrome [SS; (A) ] and mycosis fungoides [MF; (B) ], showing CXCL13 expression in endothelial cells and neoplastic lymphocytes infiltrating the dermis, frequently displaying a dot-like staining pattern. In atopic dermatitis [AD; (C) ], psoriasis [PS; (D) ], eczema [EC; (E) ] and healthy donor skin [HD; (F) ], CXCL13 immunoreactivity is mainly confined to endothelial cells and scattered non-neoplastic lymphocytes. Sections were counterstained with haematoxylin. Original magnification: ×10; inserts ×40.

Article Snippet: Sections were then incubated overnight at 4 °C with a goat polyclonal anti-CXCL13 antibody (AF801; R&D Systems) at a concentration of 2.5 μg/mL in 2% bovine serum albumin (BSA).

Techniques: Immunohistochemical staining, Expressing, Staining

Journal: Frontiers in Immunology

Article Title: CXCL13 as a simple and promising blood biomarker for differentiating Sézary syndrome from mycosis fungoides and other confounding chronic inflammatory skin diseases

doi: 10.3389/fimmu.2026.1804103

Figure Lengend Snippet: Plasma CXCL13 levels and diagnostic performance in Sézary syndrome. (A) Box-and-whisker plots showing individual plasma CXCL13 concentrations in patients with Sézary syndrome (SS), mycosis fungoides (MF), atopic dermatitis (AD), psoriasis (PS), eczema (EC) and healthy donors (HD). Statistical significance was assessed using the Kruskal–Wallis test followed by Dunn’s multiple comparisons test. (B) Plasma CXCL13 concentrations in MF patients stratified according to disease stage at sampling (stage IB/IIA vs . advanced stages IIB and III). Statistical significance was assessed using the Mann–Whitney U test. (C) Receiver operating characteristic (ROC) curve analysis evaluating the ability of plasma CXCL13 to discriminate SS patients from all non-SS individuals (MF, AD, PS, EC and HD). The red cross indicates the selected cut-off value (151.1 pg/mL) maximising sensitivity (87%) and specificity (88%). (D) Box-and-whisker plots showing individual plasma CXCL13 concentrations in SS (right) and non-SS (left) patients. The dotted line indicates the ROC-derived cut-off value. Statistical significance was assessed using the Mann–Whitney U test. ****p ≤ 0.0001; **p ≤ 0.01; *p ≤ 0.05.

Article Snippet: Sections were then incubated overnight at 4 °C with a goat polyclonal anti-CXCL13 antibody (AF801; R&D Systems) at a concentration of 2.5 μg/mL in 2% bovine serum albumin (BSA).

Techniques: Clinical Proteomics, Diagnostic Assay, Whisker Assay, Sampling, MANN-WHITNEY, Derivative Assay

Journal: The Journal of Clinical Investigation

Article Title: Microenvironmental network of clonal CXCL13 + CD4 + T cells and Tregs in pemphigus chronic blisters

doi: 10.1172/JCI166357

Figure Lengend Snippet: ( A ) Bulk RNA-Seq of tertiary lymphoid structure–positive (TLS-positive) and –negative samples ( n = 5 each). ( B ) Gene ontology (GO) analysis and ( C ) volcano plot depicting upregulated (red dots) and downregulated DEGs (blue dots). ( D ) Representative immunofluorescence staining for CXCL13 (green) and CD20 (red) in skin TLSs from a patient with pemphigus. Nuclei were stained with DAPI (light gray). Scale bar: 100 μm. ( E ) Representative immunofluorescence staining for coexpression of CD4 (red) and CXCL13 (green) in skin TLSs. White arrowheads indicate CXCL13 + CD4 + cells. Nuclei were stained with DAPI (light gray). Scale bar: 50 μm. ( F ) Percentage of CD4 + , CD8 + , CD20 + , CD138 + , FDC + , and HLA-DR + cells in CXCL13 + cells from immunofluorescence images ( n = 5). Data are shown as the mean ± SD. ( G ) Gene set enrichment analysis of Th1, Th2, Th17, and Tfh cell gene signatures using the transcriptome of TLS-positive versus TLS-negative samples. ( H ) Percentage of top 10 most frequently occurring clones of TCRβ chain from bulk TCR-Seq in skin TLSs from 5 patients (no. 1–5).

Article Snippet: Tissues were stained overnight at 4°C using the following primary antibodies: mouse anti-human CD20 (L26, Abcam); 6X His-recombinant human DSG1 and 6X His-recombinant human DSG3 (both from Cusabio); goat anti-human CXCL13 (R&D Systems); rabbit anti-human CCL5 (P230E, Thermo Fisher Scientific); rabbit anti-human CD138 (EP201), rabbit anti-human FDC (CNA.42), and mouse anti-human CD8 (C8/144B) (all from Cell Marque); rat anti-human CD4 (YNB 46.1.8, Santa Cruz); and mouse anti-human HLA-DR (L243, BioLegend).

Techniques: RNA Sequencing, Immunofluorescence, Staining, Clone Assay

Journal: The Journal of Clinical Investigation

Article Title: Microenvironmental network of clonal CXCL13 + CD4 + T cells and Tregs in pemphigus chronic blisters

doi: 10.1172/JCI166357

Figure Lengend Snippet: ( A ) Schematic of scRNA-Seq and scTCR-Seq for skin lesions with tertiary lymphoid structures (TLSs) in patients with pemphigus ( n = 4). ( B ) UMAP visualization of 2,141 TCRβ + T cells. ( C ) Violin plot showing the expression of the indicated marker genes in T cell subsets by scRNA-Seq. CD4, CD4 + T cells; CD8, CD8 + T cells. ( D ) Heatmap of the Morisita-Horn index quantifying overlapping TCRs among clusters. ( E ) The Shannon entropy calculation of the diversity of the TCR repertoire in each cluster. One-way ANOVA and Student’s t tests were used to compare means for 2 groups. ** P < 0.005; *** P < 0.0001. Data are shown as the mean ± SD. ( F ) UMAP visualization of shared TCRs between T cells in skin TLSs and DSG3-specific, activation-induced marker (AIM) + Tfh and non-Tfh memory CD4 + T cells in PBMCs from a patient with pemphigus vulgaris. ( G ) Volcano plot showing upregulated (orange dots) and downregulated (purple dots) in cluster 5 DEGs compared with cluster 1. ( H ) UMAP visualization showing gene signature of tissue-resident memory T (Trm) cells. ( I ) Dot plot showing expression of genes in the categories of costimulatory or coinhibitory receptors and glycolysis in each cluster. ( J ) Linear regression analyses of the expression of CXCL13 and the correlation with expression of TNFRSF18 , TPI1 , PGAM1 , LAG3 , and TIGIT in clusters 1 and 5. Pearson’s correlation analysis was used to measure the strength of relationships between variables. ( K ) Gene ontology analysis in cluster 5 compared with cluster 1. ( L ) Dot plot depicting the TCR-mediated gene set in each cluster. ( M ) Linear regression analysis of the correlation between CXCL13 and LCK in clusters 1 and 5. Pearson’s correlation analysis was used to measure the strength of relationships between variables.

Article Snippet: Tissues were stained overnight at 4°C using the following primary antibodies: mouse anti-human CD20 (L26, Abcam); 6X His-recombinant human DSG1 and 6X His-recombinant human DSG3 (both from Cusabio); goat anti-human CXCL13 (R&D Systems); rabbit anti-human CCL5 (P230E, Thermo Fisher Scientific); rabbit anti-human CD138 (EP201), rabbit anti-human FDC (CNA.42), and mouse anti-human CD8 (C8/144B) (all from Cell Marque); rat anti-human CD4 (YNB 46.1.8, Santa Cruz); and mouse anti-human HLA-DR (L243, BioLegend).

Techniques: Expressing, Marker, Activation Assay

Journal: The Journal of Clinical Investigation

Article Title: Microenvironmental network of clonal CXCL13 + CD4 + T cells and Tregs in pemphigus chronic blisters

doi: 10.1172/JCI166357

Figure Lengend Snippet: ( A ) Workflow for CODEX imaging and analysis of skin tertiary lymphoid structures (TLSs) in patients with pemphigus ( n = 9). ( B ) Six representative markers (left) for FoxP3 (red), CD4 (magenta), CD20 (cyan), HLA-DR (green), CXCL13 (white), and CD8 (blue) and a representative Voronoi diagram (right) of the TLSs after cell mapping. ( C ) Frequency of PD-1 + and ICOS + cells in CXCL13 + versus CXCL13 – CD4 + Tm cells. Paired t tests were used to compare values for 2-variable plots. * P < 0.05. ( D ) Densities of Tregs, HLA-DR + cells, CD8 + T cells, and B cells based on their distance from the center of CXCL13 + versus CXCL13 – CD4 + Tm cells. Wilcoxon matched-pairs signed-rank test. * P < 0.05. ( E ) Frequencies of Tregs, HLA-DR + cells, CD8 + T cells, and B cells adjacent to CXCL13 + versus CXCL13 – CD4 + Tm cells in TLSs. Paired t tests were used to compare values for 2-variable plots. * P < 0.05. ( F ) Representative figures highlighting CXCL13 + CD4 + Tm cells (red) and Tregs (yellow) in the Voronoi diagram. ( G ) Frequencies of marker-positive cells in Tregs adjacent to CXCL13 + versus CXCL13 – CD4 + Tm cells. Data are shown as the mean ± SD.

Article Snippet: Tissues were stained overnight at 4°C using the following primary antibodies: mouse anti-human CD20 (L26, Abcam); 6X His-recombinant human DSG1 and 6X His-recombinant human DSG3 (both from Cusabio); goat anti-human CXCL13 (R&D Systems); rabbit anti-human CCL5 (P230E, Thermo Fisher Scientific); rabbit anti-human CD138 (EP201), rabbit anti-human FDC (CNA.42), and mouse anti-human CD8 (C8/144B) (all from Cell Marque); rat anti-human CD4 (YNB 46.1.8, Santa Cruz); and mouse anti-human HLA-DR (L243, BioLegend).

Techniques: Imaging, Marker

Journal: The Journal of Clinical Investigation

Article Title: Microenvironmental network of clonal CXCL13 + CD4 + T cells and Tregs in pemphigus chronic blisters

doi: 10.1172/JCI166357

Figure Lengend Snippet: ( A – F ) CXCL13 + CD4 + T cells were differentiated in with and without Treg conditions in vitro. Conventional T cells were stained with CellTrace Far Red (CTFR), and sorted CD25 + CD127 lo CD4 + Tregs were stained with Cell Trace Violet (CTV). ( A ) Representative plots and ( B ) graph of the relative frequencies of CXCL13 + cells in CTFR + CD4 + and CTFR + GITR + CD4 + T cells ( n = 8). Paired t tests were used to compare values for 2-variable plots. *** P < 0.0001. ( C ) Frequencies of CTFR + conventional T cells and CTV + Tregs in CXCL13 + GITR + CD4 + T cells. ( D ) Gene ontology analysis using downregulated DEGs and ( E ) gene set enrichment analysis of IL-2 pathway gene signatures from the bulk RNA-Seq of CD4 + T cells in the Treg-undepleted condition compared with the Treg-depleted condition ( n = 5). ( F ) Dot plot showing expression of genes involved in the IL-2 pathway, TGF-β pathway, and IL-10 pathway in each cluster, as assessed by scRNA-Seq. ( G ) CXCL13 + CD4 + T cells were differentiated in the presence or absence of neutralizing anti–IL-2 antibody and/or TGF-β. Relative frequencies of CXCL13 + cells in GITR + CD4 + T cells ( n = 5). Paired t tests were used to compare values for 2-variable plots. * P < 0.05. ( H ) Differentiated CXCL13 + CD4 + T cells were cocultured with or without induced Tregs the in presence or absence recombinant IL-2 protein and TGF-β–blocking antibody. Relative frequencies of CXCL13 + cells in GITR + CD25 –/lo CD4 + T cells ( n = 8). Paired t tests were used to compare values for 2-variable plots. ** P < 0.005.

Article Snippet: Tissues were stained overnight at 4°C using the following primary antibodies: mouse anti-human CD20 (L26, Abcam); 6X His-recombinant human DSG1 and 6X His-recombinant human DSG3 (both from Cusabio); goat anti-human CXCL13 (R&D Systems); rabbit anti-human CCL5 (P230E, Thermo Fisher Scientific); rabbit anti-human CD138 (EP201), rabbit anti-human FDC (CNA.42), and mouse anti-human CD8 (C8/144B) (all from Cell Marque); rat anti-human CD4 (YNB 46.1.8, Santa Cruz); and mouse anti-human HLA-DR (L243, BioLegend).

Techniques: In Vitro, Staining, RNA Sequencing, Expressing, Recombinant, Blocking 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: 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: Primary polyclonal goat anti-human CXCL13 (cat. no. AF801, 1:20; R&D Systems) antibodies were applied and incubated in a humidified box overnight at 4°C.

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

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: Primary polyclonal goat anti-human CXCL13 (cat. no. AF801, 1:20; R&D Systems) antibodies were applied and incubated in a humidified box overnight at 4°C.

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: Primary polyclonal goat anti-human CXCL13 (cat. no. AF801, 1:20; R&D Systems) antibodies were applied and incubated in a humidified box overnight at 4°C.

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: Primary polyclonal goat anti-human CXCL13 (cat. no. AF801, 1:20; R&D Systems) antibodies were applied and incubated in a humidified box overnight at 4°C.

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: Primary polyclonal goat anti-human CXCL13 (cat. no. AF801, 1:20; R&D Systems) antibodies were applied and incubated in a humidified box overnight at 4°C.

Techniques: Migration, Expressing, Concentration Assay, Modification