Additional file 1 of Peripheral immune cell dysregulation following diffuse traumatic brain injury in pigs
Creators: name: Wofford, Kathryn L.
name: Browne, Kevin D.
name: Loane, David J.
name: Meaney, David F.
name: Cullen, D. Kacy
Publisher: name: figshare
Year: 2024
Subjects: subject: Immunology
subject: FOS: Clinical medicine

description: Supplementary Material 1: Supp. Figure 1. Flow cytometry gating strategy to characterize PBMCs. Samples were gated to exclude debris (A), doublets (B), and dead cells (C). Live cells were then gated on CD52 and CD172 to segregate myeloid and lymphoid cell populations (D). Lymphoid cells were separated into CD3 + T cells or CD3- NK cells (E). CD3 + T cells were gated for CD4a and CD8a signals (F). Blue, red, and orange populations are FMO or unstained controls. Supp. Figure 2. Changes to circulating immune cell demographics over time after a sham or a moderate rotational velocity TBI. Representative H&amp;E images after a sham (A) or moderate rotational velocity TBI (F). The circulating PBMC concentration (B, G), myeloid cell fraction (C, H), T cell fraction (D, I), and NK cell fraction (E, J) after a sham or a moderate rotational velocity TBI. Supp. Figure 3. Percent change in circulating myeloid, T, and NK cells. Circulating PBMCs were collected after a high rotational velocity TBI (A), moderate rotational velocity TBI (B), or a sham procedure (C). Repeated measures subtracted pre-injury baseline levels to reduce animal-to-animal variability in myeloid cells, T cells, and NK cell populations. Supp. Figure 4. T cell subtype dynamics over time after a sham or a moderate rotational velocity TBI. T cell subsets over time after a sham (A) or moderate rotational velocity TBI (F). The percentage of circulating CD8+ (B, G), CD4+ (C, H), double positive (CD8+/CD4+; D, I), and double negative (CD8−/CD4−; E, J) T cell subset after a sham or a moderate rotational velocity TBI. Supp. Figure 5. The percentage of T cell subtypes within T cells. The percentage of circulating CD8+, CD4+, double positive (CD8+/CD4+), and double negative (CD8−/CD4−) cells within the CD3 + T cells after a high rotational velocity TBI (A), moderate rotational velocity TBI (B), or sham procedure (C). Supp. Figure 6. CD8 to CD4 T cell ratios. The ratio of CD8 to CD4 T cells over time across sham (A), moderate rotational velocity TBI (B), or high rotational velocity TBI (C). Supp. Figure 7. ROS production from stimulated PBMCs. ROS production from stimulated PBMCs that were extracted over time after a sham (A), moderate rotational velocity TBI (B), or high rotational velocity TBI (C). Supp. Figure 8. Phagocytosis changes over time. Changes in phagocytosis uptake across targets (A) of extracted PBMCs after a sham procedure (B) or a moderate rotational velocity TBI (C). Supp. Figure 9. Target-loaded PBMC population changes over time. Polystyrene particles (A) were phagocytosed by PBMCs at all timepoints (B). The number of bead + cells (C) and the fluorescence of the bead + cell population (D) trends after a high rotational velocity TBI. Supp. Figure 10. Changes to cytokines over time. Plasma cytokine levels over time following a sham, moderate rotational velocity TBI, or high rotational velocity TBI. Supp. Figure 11. Changes to spleen and thymus composition across injuries. Outcome measures for the spleen (A-F) and the thymus (G-L) were not significantly different across injury velocity. Sham and high rotational velocity TBI data is repeated from Figs. 7 and 8 to provide context for moderate rotational velocity TBI outcomes.