anti cxcl16 neutralizing antibody  (R&D Systems)

Journal: Genome Medicine

Article Title: Single-cell RNA transcriptome analysis of CNS immune cells reveals CXCL16/CXCR6 as maintenance factors for tissue-resident T cells that drive synapse elimination

doi: 10.1186/s13073-022-01111-0

Figure Lengend Snippet: Clustering of CNS cells following WNV infection by scRNA-seq. a Experimental design for scRNA-sequencing. Mice were infected (i.c.) with 1×10 4 p.f.u. WNV-NS5-E218A and harvested 25 DPI. N= 2 per group, each N includes 4 mice pooled. b tSNE plot of all immune cells analyzed in both mock- and WNV-infected animals, showing 9 clusters, colored by density clustering, and annotated by cell-type identity and number of cells in parentheses. c tSNE plots of all cells analyzed in mock (left panel)- and WNV (right panel)-infected mice, separated by treatment group, colored by density clustering. d Heatmap of single cells representing the mRNA levels of the top three well-known genes used for cellular identification of each cluster. e tSNE plot of all immune cells with mRNA of Cxcr6 (left panel) and mRNA of Cxcl16 (right panel). f Violin plots showing immune cells with Cxcr6 mRNA (left panel) or Cxcl16 mRNA (right panel) in clusters 0–5. Each dot represents a cell

Article Snippet: Monoclonal rat IgG 2A CXCL16 neutralizing antibody (R&D Systems, Clone # 142417, cat. no. MAB503) or monoclonal Rat IgG 2A isotype control (R&D Systems, Clone # 54447 cat. no. MAB006) was administered via retro-orbital injection at a dose of 25μg for 4 consecutive days on days 7, 8, 9, and 10 post-infection.

Techniques: Infection, Sequencing

Journal: Genome Medicine

Article Title: Single-cell RNA transcriptome analysis of CNS immune cells reveals CXCL16/CXCR6 as maintenance factors for tissue-resident T cells that drive synapse elimination

doi: 10.1186/s13073-022-01111-0

Figure Lengend Snippet: CXCL16 levels increase on IBA1 + cells during acute infection and return to baseline levels during recovery. a Representative microscopy images of RNA in situ/immunohistochemistry (ISH/IHC) for Cxcl16 and IBA1 in the cortex of mock- or WNV-infected mice at 7, 25, ad 52 DPI. The middle panel is the inset of the white box in 7 DPI. b – d Quantification of the ISH/IHC for Cxcl16 + area ( b ); IBA1 + Cxcl16 + area normalized to the total IBA1 + area, represented as fold change over mock ( c ); or IBA1 + Cxcl16 + area normalized to the total Cxcl16 + area ( d ) in the cortex or hippocampus of mock- or WNV-infected mice at 7, 25, and 52 DPI. e ELISA for CXCL16 in the cortex, hippocampus, cervical lymph nodes, meninges, and blood at 7 and 25 DPI. f Representative immunostaining of the cortex for CXCR6-GFP, CD3, IBA1, and DAPI. Scale bars, 50 μm. Data represent the mean±s.e.m. and were analyzed by one-way ANOVA or unpaired Student’s t -test. * P <0.05, ** P <0.005, *** P < 0.001, **** P < 0.0001

Article Snippet: Monoclonal rat IgG 2A CXCL16 neutralizing antibody (R&D Systems, Clone # 142417, cat. no. MAB503) or monoclonal Rat IgG 2A isotype control (R&D Systems, Clone # 54447 cat. no. MAB006) was administered via retro-orbital injection at a dose of 25μg for 4 consecutive days on days 7, 8, 9, and 10 post-infection.

Techniques: Infection, Microscopy, In Situ, Immunohistochemistry, Enzyme-linked Immunosorbent Assay, Immunostaining

Journal: Genome Medicine

Article Title: Single-cell RNA transcriptome analysis of CNS immune cells reveals CXCL16/CXCR6 as maintenance factors for tissue-resident T cells that drive synapse elimination

doi: 10.1186/s13073-022-01111-0

Figure Lengend Snippet: CXCL16 neutralization leads to decreased percentage of T R M cells in the CNS after viral clearance. a . Schematic depicting experimental design for CXCL16 neutralizing an-tibody experiment. Mice were infected (i.c.) with 1×10 4 p.f.u. WNV-NS5-E218A and administered CXCL16 antibody or isotype control via retro-orbital injection, and harvested 15 DPI. b . Quantifi-cation of percentage of CD45 high cells that are CD8 + in the cortex (left) and hippocampus (right) in mice that received isotype or CXCL16 neutralizing antibody. c . Gating strategy and quantifi-cation of percentage of CD8 + T cells that are CD103 + in the cortex and hippocampus in mice that received isotype or CXCL16 neutralizing antibody. Data represent the mean±s.e.m. and were analyzed by unpaired Student’s t -test. *P<0.05

Article Snippet: Monoclonal rat IgG 2A CXCL16 neutralizing antibody (R&D Systems, Clone # 142417, cat. no. MAB503) or monoclonal Rat IgG 2A isotype control (R&D Systems, Clone # 54447 cat. no. MAB006) was administered via retro-orbital injection at a dose of 25μg for 4 consecutive days on days 7, 8, 9, and 10 post-infection.

Techniques: Neutralization, Infection, Control, Injection

Journal: The FASEB Journal

Article Title: Pathogenic Tconvs promote inflammatory macrophage polarization through GM‐CSF and exacerbate abdominal aortic aneurysm formation

doi: 10.1096/fj.202101576R

Figure Lengend Snippet: The CXCL16/ CXCR6 axis mediates the recruitment of Tconvs in abdominal aortic aneurysm (AAA). (A) Scatter plots comparing gene expression quantified by RNA‐sequencing of aortic aneurysm Tconvs versus splenic Tconvs. The Cxcr6 gene is highlighted in red. (B) Representative flow cytometry plots (left) and quantification (right) of CXCR6 expression of Tconvs in aortic aneurysms and spleens 7 days after AAA induction. n = 5 per group. **** p < .0001 according to unpaired two‐tailed t test. (C) CXCL16 transcripts in aortic aneurysms were quantified by RT‐PCR on day 0, day 1, day 3, day 7, and day 14 after AAA induction. n = 8–12 per group. * p < .05, *** p < .001, **** p < .0001 versus day 0 according to Kruskal–Wallis test. (D) CXCL16 protein levels in aortic aneurysms were quantified by western blotting at day 0, day 1, day 3, day 7, and day 14 after AAA induction. Left: representative immunoblot. Right: summary data. n = 6 per group. ** p < .01, *** p < .001, **** p < .0001 versus day 0 according to one‐way ANOVA. (E) Proportion (left) and number (right) of Tconvs in aortic aneurysms after treatment with IgG (control) or CXCL16 antibody. n = 4–5 per group. * p < .05 according to Mann–Whitney test on proportion and unpaired two‐tailed t test on number. (F) Representative flow cytometry plots (left) and quantification (right) of CXCR6 + Tconvs in aortic aneurysms. n = 4–5 per group. * p < .05 according to unpaired two‐tailed t test. (G) Proportion (left) and number (right) of Tconvs in aortic aneurysms of CXCR6 GFP/+ and CXCR6 GFP/GFP mice at day 7 after AAA induction. n = 6 per group. * p < .05, ** p < .01 according to unpaired two‐tailed t test. (H) Representative pictures (left) of abdominal aortic fragments and quantification of maximal diameters (right) from CXCR6 GFP/+ and CXCR6 GFP/GFP mice 2 weeks after porcine pancreatic elastase –induced AAA. n = 6 per group. *** p < .001 according to unpaired two‐tailed t test

Article Snippet: To block CXCL16, mice were treated with intraperitoneal injections of 100 μg of monoclonal rat anti‐mouse CXCL16 neutralizing antibody every 3 days (R&D Systems, Clone #142417, Germany), and the control group was treated with an isotype control Ab.

Techniques: Gene Expression, RNA Sequencing, Flow Cytometry, Expressing, Two Tailed Test, Reverse Transcription Polymerase Chain Reaction, Western Blot, Control, MANN-WHITNEY