anti vla 4  (Bio X Cell)

Journal: bioRxiv

Article Title: Blockade of VCAM1 or VLA4 preserves cerebrovasculature and prevents cognitive decline late after stroke

doi: 10.1101/2025.06.25.661593

Figure Lengend Snippet: A) Escape latency on the Barnes maze and B) discrimination index on the novel object recognition task for 10 month old male mice treated with isotype control antibody (α-IC; n=10 sham and 11 stroke), anti-VCAM1 (α-VCAM1; n=11) or anti-VLA4 (α-VLA4; n=10) chronically. Statistics, 2-way repeated measures ANOVA with Bonferroni’s post-hoc test; Error bars, mean ± SEM; *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001.

Article Snippet: Adult (3 month old) and middle-aged (10 month old) C57BL/6J male and female mice received injections of anti-VCAM1 monoclonal antibody (α-VCAM1, BE0027, BioXCell), anti-VLA4 monoclonal antibody (α-VLA4, BE0071, BioXCell) or the appropriate isotype control antibody (α-IC, VCAM1: BE0088; VLA4: BE0090; BioXCell) at a dose of 9 mg per kg. For experiments with an acute dosing group, the first dose at 4 hours after surgery was retro-orbital, and subsequent doses were intraperitoneal.

Techniques: Control

Journal: bioRxiv

Article Title: Blockade of VCAM1 or VLA4 preserves cerebrovasculature and prevents cognitive decline late after stroke

doi: 10.1101/2025.06.25.661593

Figure Lengend Snippet: A) Quantification and representative bright field images of CD3+ T cells in the stroke core after isotype control (α-IC, n=8), anti-VCAM1 acute (Stroke-α-VCAM1 Acute, n=9) or chronic (Stroke-α-VCAM1 Chronic, n=9) treatment, or B) isotype control (α-IC, n=7), anti-VLA4 acute (Stroke-α-VLA4, n=5) or chronic (Stroke-α-VLA4 Chronic, n=8) antibody treatment 3 weeks after stroke. C-D) Quantification and representative bright field images of B220+ B cells in the stroke core after in the same mice. Scale bar, 300 µM (top row) or 50 µM (bottom row); statistics, 2-way ANOVA with Tukey’s post hoc test for group comparisons; Error bars, mean ± SEM; *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001.

Article Snippet: Adult (3 month old) and middle-aged (10 month old) C57BL/6J male and female mice received injections of anti-VCAM1 monoclonal antibody (α-VCAM1, BE0027, BioXCell), anti-VLA4 monoclonal antibody (α-VLA4, BE0071, BioXCell) or the appropriate isotype control antibody (α-IC, VCAM1: BE0088; VLA4: BE0090; BioXCell) at a dose of 9 mg per kg. For experiments with an acute dosing group, the first dose at 4 hours after surgery was retro-orbital, and subsequent doses were intraperitoneal.

Techniques: Control

Journal: bioRxiv

Article Title: Blockade of VCAM1 or VLA4 preserves cerebrovasculature and prevents cognitive decline late after stroke

doi: 10.1101/2025.06.25.661593

Figure Lengend Snippet: Anti-VCAM1 and anti-VLA4 treatment induces changes in endothelial cells which reflect generation of new and more mature blood vessels. Unbiased UMAP visualization of (A) immune and (B) endothelial cell populations isolated from the stroke scar of 10 month old male mice, 10 weeks after stroke, based on single cell RNA sequencing. Each sample represents a pool of 4-5 mice in four groups, sham and stroke mice treated with isotype control antibody (Sham-α-IC and Stroke-α-IC), and stroke mice treated chronically with anti-VCAM1 (Stroke-α-VCAM1) or anti-VLA4 (Stroke-α-VLA4) antibody. For all groups, treatment was every 3 days, beginning 4 days after surgery for a total of 10 weeks. C) A graphical representation of the number of differentially expressed genes in each subpopulation of endothelial cells, separated by the specific comparisons of interest. D-G) Over-representation pathway analysis on the differentially expressed genes in the artery and capillary-1 clusters. Comparisons are between Stroke-IgG and either Stroke-VCAM1 (D,F) or Stroke-VLA4 (E,G) treatment groups. The top pathways are included here, along with the combined score (x-axis), the percent of genes expressed within each pathway (dot size) and p value of pathway enrichment (dot color). Blue arrows identify pathways involved in vessel growth, while red arrows identify pathways involved in vessel maturation and blood brain barrier maintenance.

Article Snippet: Adult (3 month old) and middle-aged (10 month old) C57BL/6J male and female mice received injections of anti-VCAM1 monoclonal antibody (α-VCAM1, BE0027, BioXCell), anti-VLA4 monoclonal antibody (α-VLA4, BE0071, BioXCell) or the appropriate isotype control antibody (α-IC, VCAM1: BE0088; VLA4: BE0090; BioXCell) at a dose of 9 mg per kg. For experiments with an acute dosing group, the first dose at 4 hours after surgery was retro-orbital, and subsequent doses were intraperitoneal.

Techniques: Isolation, RNA Sequencing, Control

Journal: bioRxiv

Article Title: Blockade of VCAM1 or VLA4 preserves cerebrovasculature and prevents cognitive decline late after stroke

doi: 10.1101/2025.06.25.661593

Figure Lengend Snippet: A) Heat map comparing the Log 2 Fold Change of selected genes involved in cell-cell junction formation, pericyte recruitment or cytoskeleton from each endothelial cell cluster. Three different bioinformatic comparisons are depicted from left to right: Stroke-α-IC v. Sham-α-IC; Stroke-α-VCAM1 v. Stroke-α-IC; Stroke-α-VLA4 v Stroke-α-IC. A: Artery cell cluster; C1: Capillary-1 cluster; C2: Capillary-2 cluster; V1: Vein-1 cluster; V2: Vein-2 cluster. A star represents a significant Log 2 Fold Change (univariate p < 0.05), and a circle represents a change >10% of cells expressing the selected gene. Open circles represent decrease in the percent of cells expressing that gene, while filled circles represent an increase. B-D) Normalized gene expression (scaled transcript counts and percent of population expressing) of selected genes involved in cell-cell junction formation (B; Ctnnb1 ), pericyte recruitment (C; Edn3 ) and blood-brain barrier maintenance (D; Plod2 ) in the capillary-1 cell cluster, and how this changes with treatment. Dot size represents the % of cells within each clustering expressing the individual gene. E-G) Protein concentration from SomaLogic proteomics of Catenin B (E), Endothelin 3 (F), and Procollagen Lysyl Hydroxylase 2 (G). * p < 0.05, Mann-Whitney U test.

Article Snippet: Adult (3 month old) and middle-aged (10 month old) C57BL/6J male and female mice received injections of anti-VCAM1 monoclonal antibody (α-VCAM1, BE0027, BioXCell), anti-VLA4 monoclonal antibody (α-VLA4, BE0071, BioXCell) or the appropriate isotype control antibody (α-IC, VCAM1: BE0088; VLA4: BE0090; BioXCell) at a dose of 9 mg per kg. For experiments with an acute dosing group, the first dose at 4 hours after surgery was retro-orbital, and subsequent doses were intraperitoneal.

Techniques: Expressing, Gene Expression, Protein Concentration, MANN-WHITNEY

Journal: bioRxiv

Article Title: Blockade of VCAM1 or VLA4 preserves cerebrovasculature and prevents cognitive decline late after stroke

doi: 10.1101/2025.06.25.661593

Figure Lengend Snippet: A) Total vessel length in the peri-infarct cortex after 3 weeks of chronic isotype control (Stroke-α-IC) or anti-VCAM1 (Stroke-α-VCAM1) antibody treatment, using endothelial staining for CD31 to define vasculature. B) Representative confocal images and (C) quantification of pericyte (CD13+) coverage of CD31+ vasculature in the peri-infarct cortex or contralateral cortical region of stroked mice treated with isotype control (Stroke-α-IC) or anti-VCAM1 (Stroke-α-VCAM1) antibody. D) Total vessel length in the peri-infarct cortex after 3 weeks of chronic isotype control (Stroke-α-IC) or anti-VLA4 (Stroke-α-VLA4) antibody treatment. E) Representative confocal images and (F) quantification of pericyte (CD13+) coverage of CD31+ vasculature in the peri-infarct cortex or contralateral cortical region of stroked mice treated with isotype control (Stroke-α-IC) or anti-VLA4 (Stroke-α-VLA4) antibody. Scale bar, 20 µM; Statistics, Student’s t test or 2-way ANOVA with Tukey’s post hoc test for group comparisons; Error bars, mean ± SEM; *p < 0.05; **p < 0.01.

Article Snippet: Adult (3 month old) and middle-aged (10 month old) C57BL/6J male and female mice received injections of anti-VCAM1 monoclonal antibody (α-VCAM1, BE0027, BioXCell), anti-VLA4 monoclonal antibody (α-VLA4, BE0071, BioXCell) or the appropriate isotype control antibody (α-IC, VCAM1: BE0088; VLA4: BE0090; BioXCell) at a dose of 9 mg per kg. For experiments with an acute dosing group, the first dose at 4 hours after surgery was retro-orbital, and subsequent doses were intraperitoneal.

Techniques: Control, Staining

Journal: bioRxiv

Article Title: Blockade of VCAM1 or VLA4 preserves cerebrovasculature and prevents cognitive decline late after stroke

doi: 10.1101/2025.06.25.661593

Figure Lengend Snippet: A) Representative confocal images and B) quantification of the percent area covered by extravascular fibrinogen in the stroke core (top) and peri-infarct cortex (bottom) of mice treated with chronic isotype control (Stroke-α-IC) or anti-VCAM1 (Stroke-α-VCAM1) antibody for 3 weeks. C) Representative confocal images and D) quantification of the percent area covered by extravascular fibrinogen in the stroke core (top) and peri-infarct cortex (bottom) of mice treated with chronic isotype control (Stroke-α-IC) or anti-VLA4 (Stroke-α-VLA4) antibody for 3 weeks. White arrows point to extravascular fibrinogen. Scale bar, 20 µM; Statistics, Student’s t test; Error bars, mean ± SEM; *p < 0.05.

Article Snippet: Adult (3 month old) and middle-aged (10 month old) C57BL/6J male and female mice received injections of anti-VCAM1 monoclonal antibody (α-VCAM1, BE0027, BioXCell), anti-VLA4 monoclonal antibody (α-VLA4, BE0071, BioXCell) or the appropriate isotype control antibody (α-IC, VCAM1: BE0088; VLA4: BE0090; BioXCell) at a dose of 9 mg per kg. For experiments with an acute dosing group, the first dose at 4 hours after surgery was retro-orbital, and subsequent doses were intraperitoneal.

Techniques: Control

Journal: bioRxiv

Article Title: Blockade of VCAM1 or VLA4 preserves cerebrovasculature and prevents cognitive decline late after stroke

doi: 10.1101/2025.06.25.661593

Figure Lengend Snippet: The diagram depicts our proposed hypothesis of how infarct-induced neurodegeneration, a chronic process, can be prevented with chronic blockade of the VLA4/VCAM1 axis. We propose that infarct-induced neurodegeneration occurs when the initial inflammation due to stroke fails to resolve, resulting in chronic immune cell trafficking to the brain, chronic blood-brain barrier leakiness and subsequent cognitive decline. Chronic blockade of the VLA4/VCAM1 signaling axis restores blood-brain barrier integrity, reduces immune cell trafficking to the brain, and prevents chronic cognitive decline after stroke. Diagram created using BioRender.

Article Snippet: Adult (3 month old) and middle-aged (10 month old) C57BL/6J male and female mice received injections of anti-VCAM1 monoclonal antibody (α-VCAM1, BE0027, BioXCell), anti-VLA4 monoclonal antibody (α-VLA4, BE0071, BioXCell) or the appropriate isotype control antibody (α-IC, VCAM1: BE0088; VLA4: BE0090; BioXCell) at a dose of 9 mg per kg. For experiments with an acute dosing group, the first dose at 4 hours after surgery was retro-orbital, and subsequent doses were intraperitoneal.

Techniques: