mouse cxcl16 antibody  (Bio-Techne corporation)

Journal: Nature Communications

Article Title: Modeling NK-cell lymphoma in mice reveals its cell-of-origin and microenvironmental changes and identifies therapeutic targets

doi: 10.1038/s41467-024-53376-1

Figure Lengend Snippet: a UMAP plot of subclustering of myeloid cells. b The distribution of myeloid subclusters in each group in the extranodal tissues and SP revealed by R o/e analysis. +++ indicates R o/e ≥ 3; ++, 2 ≤ R o/e < 3; +, 1.5 ≤ R o/e < 2; +/−, 0.67 ≤ R o/e < 1.5; −, 0 ≤ R o/e < 0.67. c GSEA analysis of scRNA-seq data comparing each myeloid cell subcluster from pre-tumor samples (SG and SP from WT and Trp53 − / − mice) and Trp53 − / − tumor samples (top) and from Trp53 − / − and Trp53 − / − LMP1 + tumor samples (bottom). Signatures significant (FDR < 0.1) across at least two subclusters are shown. d Comparison of outgoing and incoming signals between Trp53 − / − and Trp53 − / − LMP1 + tumors in CellChat analysis. e Relative IFN-γ expressions in Lin − CD122 + cells from Trp53 − / − (n = 4) and Trp53 − / − LMP1 + (n = 3) tumors by intracellular flow cytometry. f Chord diagram of Ifng -( Ifngr1 + Ifngr2 ) (left), Cxcl16 - Cxcr6 (middle), and Cxcl9 - Cxcr3 (right) signaling network across subclusters in Trp53 − / − and Trp53 − / − LMP1 + tumors. g Relative CXCR6 (left) and CXCR3 (right) expressions in Lin − CD122 + cells from Trp53 − / − (n = 4) and Trp53 − / − LMP1 + (n = 4) tumors by flow cytometry. h CXCR6 (left) and CXCR3 (right) gene expressions in human normal NK cells (n = 4) and ENKTCL tumors (n = 41) by RNA-seq. i Representative images of CXCR6 (left) and CXCR3 (right) immunostaining in a human ENKTCL sample. j Relative cell number of Trp53 − / − and Trp53 − / − LMP1 + tumor cells 48 h after CXCL16 or CXCL9 treatment at indicated concentrations (n = 4). k The number of Trp53 − / − LMP1 + tumor cells after one week of co-culture with and without cDC (n = 6). l Kaplan–Meier survival curves of mice transplanted with 5 × 10 5 Trp53 − / − LMP1 + tumor cells and administered with anti-CXCL16 antibody (n = 6) or isotype control (n = 8). Log-rank test. m Kaplan–Meier survival curves of WT or Cxcl16 knockout mice transplanted with 5 × 10 5 Trp53 − / − LMP1 + tumor cells and administered with control vehicle or anti-KLRG1 antibody (n = 7 per group). Log-rank test. g , j lines show means. h , k Box plots show medians (lines), IQRs (boxes), and ±1.5× IQR (whiskers). e , g , h , j , and k NS: not significant, *P < 0.05, **P < 0.005, ***P < 0.0005, two-sided Welch’s t-test. No adjustments were made for multiple comparisons. e , g , h , and j – m Source data are provided as a Source Data file.

Article Snippet: Five mg/kg of anti-CXCL16 antibody (R&D Systems) or their isotype Rat IgG2A antibody (R&D Systems) was administered intraperitoneally every one week beginning from day 5 post-transplantation.

Techniques: Comparison, Flow Cytometry, RNA Sequencing Assay, Immunostaining, Co-Culture Assay, Control, Knock-Out

Journal: bioRxiv

Article Title: Nasal tissue-resident memory CD4 + T cells persist after influenza A virus infection and provide heterosubtypic protection

doi: 10.1101/2024.07.06.602325

Figure Lengend Snippet: NT CD4 TRM rely on CXCR6-CXCL16 axis for their formation A) Box plot showing the expression of Cxcr6 mRNA among CD4 TRM of the NT and lungs. Data are presented as median and interquartile range. NS, not significant, **** P <0.0001; *** P <0.001; ** P <0.01; * P <0.05 by unpaired two-tailed t-test. B) Box plot showing the expression of Cxcr6 mRNA among different cell clusters of lungs and NT. Data are presented as median and interquartile range. NS, not significant, **** P <0.0001; *** P <0.001; ** P <0.01; * P <0.05 by Wilcoxon rank sum test with Benjamini-Hochberg correction C) Volcano plot for differential expressed genes in the NT in comparison to the lungs of the Th17 cluster. The dotted lines indicate fold change and adjusted p value cutoffs. D) Bar plot for the expression of CXCR6 (each Median Fluorescence Intensity (MFI) normalized to mean MFI from CD4 TEM iv + of NT) in CD4 TRM and iv + CD4 TEM from the lungs and NT of naïve mice and PR8 IAV infected mice (30 dpi) as indicated. The experiment was repeated thrice and the results (mean ± s.e.m.) are pooled. NS, not significant, **** P <0.0001; *** P <0.001; ** P <0.01; * P <0.05 by two-way ANOVA, with Tukey’s multiple comparison test. E-F) Expression of CXCR6 on OT-II CD4 TRM of NT and lung on d30 following PR8-Ova infection as indicated. E) Representative histograms for CXCR6 expression from OT-II CD4 TRM of NT and lung are shown. F) Scatter plot showing the expression of CXCR6 (each Median Fluorescence Intensity (MFI) normalized to mean MFI of NT OT-II CD4 TRM). The experiment was repeated thrice and the results (mean ± s.e.m.) are pooled. NS, not significant, **** P <0.0001; *** P <0.001; ** P <0.01; * P <0.05 by unpaired two- tailed t-test. G-J) Frequency of different cell populations within different organs derived from WT- Cxcr6 -/- BM chimeric mice on day 30 post infection with PR8 IAV as indicated. G) Schematic representation of the experimental setup. H) Representative flow cytometry plots showing the percentage of WT and Cxcr6 -/- CD4 TRM in NT and lungs. I) Scatter plot for the percentage of WT and Cxcr6 -/- CD4 TRM in NT and lungs as indicated. J) Scatter plot for the percentage of I-A b NP306-322 tetramer - specific WT and Cxcr6 -/- CD4 TRM in NT and lungs as indicated. The experiment was repeated thrice and the results (mean ± s.e.m.) are pooled. NS, not significant, **** P <0.0001; *** P <0.001; ** P <0.01; * P <0.05 by unpaired two-tailed t-test. K) A representative microscopic image of CXCL16 expression (magenta) and OT-II CD4 T cells (red and green) in the olfactory epithelium of the murine NT. NT is isolated on day 30 post PR8-Ova infection from mice that received OT-II CD4 T cells. L-N) Antigen-specific CD4 TRM in the lungs and NT of mice treated with isotype control or anti-CXCL16 antibody on day 10 post PR8 IAV infection. L) Schematic representation of the experimental setup. M) Representative flow cytometry plots indicating the percentage of I-A b NP306-322 tetramer - specific CD4 TRM are shown. N) Scatter plot indicating the absolute number of I-A b NP306-322 tetramer - specific CD4 TRM. The experiment was performed twice and the results (mean ± s.e.m.) are pooled. NS, not significant, **** P <0.0001; *** P <0.001; ** P <0.01; * P <0.05 by unpaired two-tailed t- test.

Article Snippet: Mouse CXCL16 antibody (MAB503) was purchased from R&D systems and was dissolved in 1X PBS.

Techniques: Expressing, Two Tailed Test, Comparison, Fluorescence, Infection, Derivative Assay, Flow Cytometry, Isolation, Control