anti mouse cxcl16 antibodies  (Bio X Cell)

Journal: Nature Communications

Article Title: Single cell transcriptomics of human psoriasis and epidermal specific Ube2l3 deficient mice highlight CXCL16/CXCR6 involvement in psoriasis development

doi: 10.1038/s41467-025-64106-6

Figure Lengend Snippet: a UMAP visualization of 398188 cells epidermal cells across control (CE) and Ube2l3 △Epi (UE) mice, classified into 6 immune/non-immune clusters via Louvain algorithm. Cell cluster: cyc-T, cycling T cell, Krt, keratinocytes, Mel, melanocytes, NK/T, natural kill T cells and T cells, SG, sebaceous gland. b Marker expression dot plot for the 6 cell populations. Dot size = expressing cell fraction; color intensity = mean scaled expression. c Compositional changes of epidermal cell states in CE vs. UE. d Predicted ligand-receptor interactions between clusters. Red arrow: upregulated Cxcl16 (myeloid)→ Cxcr6 (NK/T) axis in UE. e Signaling activity of CXCR6-CXCL16 and top ligand-receptor pairs across cell types. f Dot plot showing the differentially expressed Vγ2 and Vγ4 specified marker ( Tcrg-V4, 5830411N06Rik and Cd163l ) in T-cell subsets. g Flow cytometry: Increased IL-17A⁺CXCR6⁺CD3⁺ T cells in Ube2l3 △Epi mice epidermis. n = 6/group. h Flow cytometry plots and quantification of the percentage of Vγ2 and Vγ3 cells in γδT cells in Epi-Ctrl and Epi -Ube2l3 △Epi groups. Epi-Ctrl, n = 5, Epi -Ube2l3 △Epi , n = 6. i Flow cytometric analysis of the percentage of TNFa + and IL-17A + Vγ2 + cells in Vγ2 + γδT cells between Epi-Ctrl and Epi -Ube2l3 △Epi groups. Epi-Ctrl, n = 5, Epi -Ube2l3 △Epi , n = 6. j Schematic experimental design showing Ube2l3 △Epi mice treated with or without anti-Vγ2mAb (10ug/g/week) intraperitoneally in four weeks. k Representative phenotype of Ube2l3 △Epi mice treated with or without anti-Vγ2mAb before (Week 0) and after 4-week treatment (Week 4). Histological analysis of H&E staining was calculated and showed a decrease in Vγ2 antibody group. ( n = 5 per group). Immunofluorence analysis of Vγ2 (red) and CD3 (green) was showed. white triangle, double stained Vγ2 and CD3. l Proportion of L-17A + γδT cells in γδT in Epi-Ctrl, Epi -Ube2l3 △Epi and the epidermis of mouse Ube2l3 △Epi group treated with anti-Vγ2mAb (Epi- Ube2l3 △Epi - Vγ2Ab). Epi-Ctrl, n = 5, Epi -Ube2l3 △Epi , n = 6, Epi- Ube2l3 △Ep i- Vγ2Ab, n = 3. m FPKM of CXCR6 in GSE53552 (left, NL, n = 23, LS, n = 25, LS-treated, n = 20), GSE117239 (middle, NL, n = 34, LS, n = 34, LS-treated, n = 30), and GSE117239 (right, NL, n = 50, LS, n = 49, LS-treated, n = 45). n UMAP plots of CXCR6 expression in human lymphoid cells in NE and PE group. o Heatmap results showed the average expression of CXCR6 and other markers in 8 groups in human lymphoid cells. p Immunofluorescence staining of psoriatic skin for CD3 (green), CXCR6 (red), CD8 (purple), and DAPI (blue). White triangle indicated CXCR6 + and CD8 + T cells (Scale bar: 50 um). q Representative flow cytometric plots and percentage of IL-17A + CXCR6 + T cells in CD3 + T cells in Epi-HD ( n = 6) and Epi-PSO ( n = 3). The proportion of CD4 + and CD8 + in IL-17A + CXCR6 + T cells. Data are presented as mean ± SEM, and p -values were calculated by unpaired, two-tail student’s t -test ( g , h , i , k and q ), one-way ANOVA with Tukey’s multiple comparisons ( l , m ). The Figure j was created in BioRender. Chen, X. (2025) https://BioRender.com/854rwn9 .

Article Snippet: The experimental design is shown in Fig. . Anti-CXCL16: Mice received subcutaneous injections (s.c.) of 4 μg/g/week goat anti-mouse CXCL16 polyclonal antibody (AF503, R&D Systems) or goat IgG isotype control.

Techniques: Control, Marker, Expressing, Activity Assay, Flow Cytometry, Staining, Immunofluorescence

Journal: Nature Communications

Article Title: Single cell transcriptomics of human psoriasis and epidermal specific Ube2l3 deficient mice highlight CXCL16/CXCR6 involvement in psoriasis development

doi: 10.1038/s41467-025-64106-6

Figure Lengend Snippet: a The single cell profile of 28,725 Keratinocytes (Krt) about nine classes conserved epidermis across UE and CE, delineated by Louvain clustering. Basal, basal keratinocytes ( Krt14 and Krt5 ), spinous cell ( Krt1 and Krt10 ), granular cell ( Spink5 and Calm5 , Krt17 , Krt79 ), hair follicle stem cells (HFSC) ( Cd34 , Postn , Ptn and Fst ). b Proportional abundance of KC states in CE vs. UE. c Scatter plots defining KC clusters. d Top upregulated genes in krt subset in scRNA-seq of UE vs CEs. e Dot plot of cytokines/ chemokines ( Cxcl16 , Ccl20 , Il23a , Tnf and Il1b) compared in each group in Krt subset. red arrow is Prolif.cell group. f Flow cytometric plots and percentage of CXCL16 expression in CD45 - cells in Epi-Ctrl ( n = 6), Epi-IMQ ( n = 6) and Epi- Ube2l3 △Epi group ( n = 5). g The immunoblotting of UBE2L3, CXCL16 and β -actin in lysates, soluble CXCL16 (sCXCL16) in the supernatants (super.) of cultured primary mouse keratinocytes in control group (KC-Ctrl) and in Ube2l3 knockout group (KC- Ube2l3 △Epi ). h Quantification of ( g ). n = 3 biological replicates. i The immunoblotting results of UBE2L3, Pro-IL1β and β -actin in lysates, IL-1β p35 and IL-1β p17 in supernatants of KC-Ctrl and KC- Ube2l3 △Epi . j Quantification of ( i ). n = 3 biological replicates. k Mouse control keratinocytes were stimulated with IL-1β (100 ng/ml), IL-17A (100 ng/ml) for 24 hours. Immunoblotting of STAT3, pSTAT3, CXCL16 and β -actin was carried out. l Quantification of ( k ). ( n = 3 biological replicates). m The R&D Luminex assay showed that the protein level of CXCL16 was increased in Epi- Ube2l3 △Epi group. n = 5. n , o Flow cytometric plots and percentage of TNFα and CXCL16 in the epidermis of healthy donors (Epi-HD, n = 3) and psoriatic patients (Epi-PSO, n = 3). p , q Normal human epidermal keratinocytes (NHKEs) were stimulated with rhIL1β (100 ng/ml) and rhIL-17A(100 ng/ml) for 24 hours and the STAT3, pSTAT3 and CXCL16 in lysates were measured by western blot. ( n = 3 biological replicates). Data are presented as mean ± SEM, and p -values were calculated by unpaired, two-tail student’s t -test ( h , j , m , and o ), one-way ANOVA with Tukey’s multiple comparisons ( m , q ).

Article Snippet: The experimental design is shown in Fig. . Anti-CXCL16: Mice received subcutaneous injections (s.c.) of 4 μg/g/week goat anti-mouse CXCL16 polyclonal antibody (AF503, R&D Systems) or goat IgG isotype control.

Techniques: Expressing, Western Blot, Cell Culture, Control, Knock-Out, Luminex

Journal: Nature Communications

Article Title: Single cell transcriptomics of human psoriasis and epidermal specific Ube2l3 deficient mice highlight CXCL16/CXCR6 involvement in psoriasis development

doi: 10.1038/s41467-025-64106-6

Figure Lengend Snippet: a UMAP of 1647 myeloid cells from control (Mouse-CE) and Ube2l3 △Epi (Mouse-UE) epidermis, clustered into 9 subsets: mast cells/basophils (MC_Bas), neutrophils (Neu), Langerhans cells (LC), macrophages (Mac), type 1 conventional DCs (cDC1), type 2 conventional DCs (cDC2). b Proportional abundance of myeloid subsets in CE vs. UE. c Expression of cytokines ( Il1b ) and chemokine ( Cxcl16 , Cxcl10 and Adam10 ) in nine defined classes. Red arrow: Cxcl16 in mDC_cDC2 cluster. d Pseoudotime analysis of Cxcl16 gene in nine cluster above in myeloid cells. Blue circles indicate the enrichment of Cxcl16 in mDC_cDC2 and Mac. e Flow cytometry and percentage of CXCL16 in mouse DC group in Epi-ctrl and Epi- Ube2l3 △Epi . Among CXCL16 + DC, the proportion of CD11b + DC (cDC2) and CD103 + DC (cDC1) in all CXCL16 + DC. n = 6. f Schematic strategy for generating Ube2l3 Epi△ - cDC -/- mice . g The phenotype of Ube2l3 Epi△ mice and Ube2l3 Epi△ - cDC -/- mice at week 0, 4, and 8. H&E staining (week 8). Epidermal thickness qualification in each group was calculated in Ube2l3 Epi△ mice ( n = 3) and Ube2l3 Epi△ - cDC -/- mice ( n = 4). Immunofluorescence of CD11C (red), K14 (green) and DAPI (blue), was done in two groups. scale bar= 50 um. h Flow cytometric plots and the percentage of IL17A + in CD3 + T, γδT and αβT in the epidermis of each group were showed. n = 3 per group. i UMAP visualization of 3237 human myeloid cells states found in human normal epidermis (NE) and human psoriatic epidermis (PE). pDC, Representative markers used in classifying mouse myeloid cells. pDC, plasmacytoid dendritic Cells. LC Langerhans cell cluster. Prolif.T, proliferated associated T cell. cDC2_mDC, type 2 DC and mature DC. Mac, macrophage. Pre-DC_cDC1, pre-DC and type 1 DC, B cell, B lymphocyte. j Heatmap of CXCL16 expression across human myeloid clusters. k Flow cytometry of IL-23p19 and CXCL16 expression in the epidermis of healthy donors (HD) and psoriatic patients (PSO). n = 3 per group. Data are presented as mean ± SEM, and p -values were calculated by unpaired, two-tail student’s t -test ( h , e , g , h , and k ). Figure f was created in BioRender. Chen, X. (2025) https://BioRender.com/riyjlc8 .

Article Snippet: The experimental design is shown in Fig. . Anti-CXCL16: Mice received subcutaneous injections (s.c.) of 4 μg/g/week goat anti-mouse CXCL16 polyclonal antibody (AF503, R&D Systems) or goat IgG isotype control.

Techniques: Control, Expressing, Flow Cytometry, Staining, Immunofluorescence

Journal: Nature Communications

Article Title: Single cell transcriptomics of human psoriasis and epidermal specific Ube2l3 deficient mice highlight CXCL16/CXCR6 involvement in psoriasis development

doi: 10.1038/s41467-025-64106-6

Figure Lengend Snippet: a The percentage of Vγ2 + γδT in γδT in Der-Ctrl and Der- Ube2l3 △Epi group. b Schematic strategy for studying the migration of Vγ2 + γδT cells with or without Cxcl16 for 24 hours. c The migrated cell was viewed by immunofluorence staining with DAPI (blue), five independent views of lower chambers of insects were calculated between Der-Ctrl ( n = 5) and Der- Ube2l3 △Epi group ( n = 5). d Next, epidermal Vγ2 + γδT cells were sorted and stimulated with rmCXCL16 for 48 hours, and flow cytometry to be carried out to analyze the IL-17A secretion percentage. e Percentage of Vγ2 + IL-17A + T in CD3 + T in isotype and rmCXCL16 group. n = 3 per group. f , g Ube2l3 △Epi mice were treated with isotype antibodies or anti-CXCL16 mAb. H&E staining of back skin and ear skin was performed. h Epidermal single cells were stimulated with or without recombinant human CXCL16 and flow cytometric analysis of IL-17A was showed. i The percentage of IL-17A + CD3 + T /CD3 + Tcells, CD8 + IL-17A + /IL-17A + T cells, CD4 + IL-17A + /IL-17A + T cells in isotype cytokine and rhCXCL16 cytokine. j In wild-type (WT) mice, intradermal injection of recombinant mouse CXCL16 (rmCXCL16) and rmIL-23 every other day for 14 days. k H&E staining of and ear skin was performed. l Skin thickness measurements (using vernier calipers) of ear skin in control ear, rmCXCL16 ear, n = 5 per group, and control ear, rmIL-23 group, n = 3 per group. m – o Flow cytometry analysis of Vγ2 + γδT cell and Vγ3 + γδT cells in γδT, IL-17A in CD3 + T cells, and CXCR6 + IL-17A + in CD3 + T cells. n = 4 per group. Data are presented as mean ± SEM, and p -values were calculated by unpaired, two-tail student’s t -test ( a , c , e , g , l ), paired, two-tail student’s t -test ( i , k , m , n , o ). The Figure d , f , h , j was created in BioRender. Chen, X. (2025) https://BioRender.com/a73juv2 .

Article Snippet: The experimental design is shown in Fig. . Anti-CXCL16: Mice received subcutaneous injections (s.c.) of 4 μg/g/week goat anti-mouse CXCL16 polyclonal antibody (AF503, R&D Systems) or goat IgG isotype control.

Techniques: Migration, Staining, Flow Cytometry, Recombinant, Injection, Control

Journal: Nature Communications

Article Title: Single cell transcriptomics of human psoriasis and epidermal specific Ube2l3 deficient mice highlight CXCL16/CXCR6 involvement in psoriasis development

doi: 10.1038/s41467-025-64106-6

Figure Lengend Snippet: A psoriasis-like lesion mouse model was generated by conditional knock out Ube2l3 in epidermis ( Ube2l3 △Epi ), which was compared with human psoriasis scRNA-seq data and facilitate cross-species comparisons of differentiation dynamics and ligand-receptor pathways in epidermis. In particular, IL-17A was regulated by CXCR6 + Vγ2 + γδT in mouse while CXCR6 + CD8 + T in human. Ube2l3 reduction in keratinocytes activated IL-1β and then promote CXCL16 expression through STAT3 signaling. CXCR6 + γδT17/ Tc17cells are prevalent within the epidermal immune microenvironment of psoriasis. Neutralization of CXCL16 inhibits the progression of psoriasis-like lesions in Ube2l3 △Epi mice. (Created in BioRender. Chen, X. (2025) https://BioRender.com/c9dlkqv ).

Article Snippet: The experimental design is shown in Fig. . Anti-CXCL16: Mice received subcutaneous injections (s.c.) of 4 μg/g/week goat anti-mouse CXCL16 polyclonal antibody (AF503, R&D Systems) or goat IgG isotype control.

Techniques: Generated, Knock-Out, Expressing, Neutralization

Journal: Journal of Neuroinflammation

Article Title: NLRP3 activation induces BBB disruption and neutrophil infiltration via CXCR2 signaling in the mouse brain

doi: 10.1186/s12974-025-03468-6

Figure Lengend Snippet: NLRP3-activated neutrophil-induced CXCL1 and CXCL2 expression reduces extracellular Claudin-5 and VE-Cadherin on brain endothelial cells. ( A ) Total RNA was isolated from neutrophils in the bone marrow of mice, and Cxcl1 , Cxcl2 , Mmp2 , and Mmp9 expression levels were analyzed using quantitative real-time PCR (qPCR). ND: not detectable. ( B ) ELISA analysis for CXCL1 and CXCL2 in the neutrophils. ( C ) Representative scatter plots and percentage of dead cells and early apoptotic cells in the brain endothelial cells. ( D ) Representative histograms and mean fluorescence intensity (MFI) of extracellular Claudin-5 (left), intracellular ZO-1 (center), and extracellular VE-Cadherin (right) in brain endothelial cells. ( E ) Experimental scheme for co-culturing neutrophils and bEnd.3 cells. Neutrophils were isolated from the bone marrow of control and active mutant mice. bEnd.3 cells were then stimulated with the isolated neutrophils in the presence or absence of anti-CXCL1 or anti-CXCL2 blocking antibodies. ( F ) Representative histograms and mean fluorescence intensity (MFI) of Claudin-5 (upper panels) and VE-Cadherin (lower panels). Data represent the results of at least three independent experiments. Mean values are shown with error bars representing the SEM. ns: non-significant. * P < 0.05, ** P < 0.01

Article Snippet: In some experiments, bEnd.3 cells were incubated with mouse neutrophils (1 × 107 cells/well) [ ] for 24 h in the presence or absence of anti-mouse CXCL1 (0.2 μg/mL, 48415, R&D Systems, MN, USA) or anti-mouse CXCL2 (2 μg/mL, 40605, R&D Systems, MN, USA) antibodies to block CXCL1 or CXCL2 [ ].

Techniques: Expressing, Isolation, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Fluorescence, Control, Mutagenesis, Blocking Assay

Journal: Journal of Neuroinflammation

Article Title: NLRP3 activation induces BBB disruption and neutrophil infiltration via CXCR2 signaling in the mouse brain

doi: 10.1186/s12974-025-03468-6

Figure Lengend Snippet: CXCL1 and CXCL2 are produced by astrocytes, pericytes, neutrophils, and endothelial cells during neuroinflammation. Immunofluorescence staining of the cerebral cortex region in coronal brain sections from ( A and B ) LPS-injected WT mice and ( C and D ) PBS-injected WT mice. Representative fluorescent images of ( A and C ) CXCL1 and ( B and D ) CXCL 2 in astrocytes, pericytes, neutrophils, and endothelial cells in the brain. Fluorescence images were acquired by staining for GFAP, NG2, LysM, or CD31 (green), CXCL1 or CXCL2 (red), and nuclei (blue). Scale bar: 10 μm. Data represent the results from the cortex region, based on the analysis of at least 15 brain slices per group across three independent experiments. Quantification of ( E ) CXCL1 and ( F ) CXCL2 levels in these cells. * P < 0.05, *** P < 0.001, **** P < 0.0001

Article Snippet: In some experiments, bEnd.3 cells were incubated with mouse neutrophils (1 × 107 cells/well) [ ] for 24 h in the presence or absence of anti-mouse CXCL1 (0.2 μg/mL, 48415, R&D Systems, MN, USA) or anti-mouse CXCL2 (2 μg/mL, 40605, R&D Systems, MN, USA) antibodies to block CXCL1 or CXCL2 [ ].

Techniques: Produced, Immunofluorescence, Staining, Injection, Fluorescence