Extended Data Fig. 6: NICIs in PNS ganglia and nerves in atherosclerosis.

a, OR/H-staining shows the presence of epineural tertiary lymphoid cluster (TLC) surrounding PvaGs in aged Apoe^−/− mice, but not in aged WT mice (dashed line, media) (n = 12 WT, 20 Apoe^−/−). b-f, Cellularity and structures of TLCs. b, TH⁺ sympathetic PvaG-TLCs harbor CD45⁺ leukocytes including CD68⁺ macrophages (arrow), CD11c⁺ MHC-II⁺ dendritic cells (arrow), CD3e⁺ T cells (open triangle), B220⁺ B cells (filled triangle), CD138⁺ plasma cells (arrow). c, Foxp3⁺ T regulatory cells (arrow), Ki67⁺ proliferating centrocytes in germinal centre (PNA⁺) (filled triangle), Ki67⁺ proliferating B cells (arrow), IgM⁺ plasma cells (filled triangle). d, PNAd⁺ high endothelial venules (HEV) (arrow), Coll-IV⁺ or Meca32⁺ blood vessels (open arrow, arrow), Lyve1⁺ lymph vessels (open triangle), ER-TR7⁺ conduits (open arrow) or ER-TR7⁺ epineurium (filled triangle) and their connection with HEVs (arrow). e, Nerve-TLCs contain CD45⁺ leukocytes including CD68⁺ macrophages, CD3e⁺ T cells (open triangle), B220⁺ B cells (filled triangle), CD138⁺ plasma cells (arrow), PNAd⁺ HEVs (arrow), Meca 32⁺ blood vessels (arrow), Lyve1⁺ lymph vessels (open triangle), and ER-TR7⁺ conduits (open arrow). f, DRG-TLCs around epineuria adjacent to spinal meninges (arrow head) contain CD68⁺ macrophages (arrow) and B220⁺ B cells (f). n = PvaG: 19 Apoe^−/−; nerves: 12 Apoe^−/−; DRGs: 7 Apoe^−/−. g, Morphometry of epineural clusters in PvaG (n = 12 WT, 20 Apoe^−/−), nerves (n = 8 WT, 12 Apoe^−/−) and DRGs (n = 5 WT, 7 Apoe^−/−) in aged mice. Each sphere represents the total number of clusters per mouse. h, Pearson correlation coefficient of PvaG-TLC sizes (TLC/PvaG area) with both plaque sizes (intima/media area) and ATLO sizes (adventitia/media area) (n = 15 PvaG-TLCs). One symbol represents the mean value of one individual variable. i, TLO stages of epineural clusters in PvaG (n = 19 Apoe^−/−), nerves (n = 12 Apoe^−/−) and DRGs (n = 7 Apoe^−/−). Each sphere represents TLO stages per tissue. j, Heatmaps of LCM-derived PvaG microarrays show differentially regulated genes in respective immuno-inflammation-related GO terms in aged WT versus Apoe^−/− PvaGs. Analyses were performed using two-sided unpaired Student’s t test. n = 5 WT PvaGs, 6 Apoe^−/− PvaGs. k,l, Quantitative comparisons of differentially expressed up-regulated genes for cytokine activity, mast cell activation, complement activation, nervous system development, and axonogenesis in WT versus Apoe^−/− PvaGs. (n = 5 WT PvaGs, 6 Apoe^−/− PvaG). Signal intensities and statistics are reported in supplementary Table 6. m, Sympathetic gene expression in LCM-derived PvaG. n = 8 PvaGs. p, Detection of CXCL13 expression in WT and Apoe^−/− PvaGs in B cell follicles (open triangle) and in PvaG neuronal cell bodies (filled triangle). o, Schematic choreographies of PvaG-TLCs, N-TLCs, and DRG-TLCs. n, Enumeration of infiltrating intraganglionic CD68⁺ macrophages, CD3e⁺ T cells, and Giemsa-stained mast cells within PvaGs, and thoraco-lumbar DRGs (i). n = PvaG: 6 WT, 8 Apoe^−/−; DRGs: 3 WT, 5 Apoe^−/−. Scale bars, 50 µm. Data are means ± s.e.m. n represents biologically independent animals. Two-sided Mann-Whitney U-test (g); Pearson bivariate correlation (h); Kruskal-Wallis H test (i); multiple unpaired t-test corrected for multiple comparisons (Bonferroni) (k,l); one way ANOVA with Bonferroni post hoc test (m); Two-sided unpaired Student´s t-test (n)

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