Extended Data Fig. 2: Modulation of lesional neutrophil counts alters lesion stability.
From: Externalized histone H4 orchestrates chronic inflammation by inducing lytic cell death
a, Models of neutropenia and neutrophilia. b, c, Quantification of lesional neutrophils in indicated treatments. n = 79 sections (ctrl IgG), n = 25 sections (anti-Ly6G), n = 27 sections (PBS), n = 58 sections (AMD3100). Two-sided unpaired t-test. d, e, Lesion volumes of mice with induced neutropenia (d) or induced neutrophilia (e). n = 10 mice (ctrl IgG), n = 9 mice (anti-Ly6G), n = 15 mice (PBS), n = 14 mice (AMD3100). Two-sided unpaired t-test. f–h, Lesion characteristics of mice with antibody-induced neutropenia (anti-Ly6G, n = 10 mice, except in (g) n = 9 mice), genetic neutropenia (Ly6gcreMcl1flox/flox, Mcl1ΔN, n = 16 mice), induced neutrophilia (AMD3100, n = 15 mice) and genetic neutrophilia (Lyz2creCxcr4flox/flox, Cxcr4ΔN, n = 13 mice) compared with respective controls (isotype IgG (n = 10 mice), Ly6gcre (n = 18 mice), vehicle (n = 15 mice) or Lyz2cre (n = 11 mice)), (dashed line). Two-sided unpaired t-test; *P < 0.05; **P < 0.01. Displayed is the quantification of lesion size (f), fibrous cap (FC) thickness (g) and collagen area (h). i, j, Analysis of SMA+MYH11+ (i) and SMA-MYH11+ (j) SMCs in indicated treatments. n = 10 mice (ctrl IgG), n = 9 mice (anti-Ly6G), n = 7 mice (PBS), n = 8 mice (AMD3100). Two-sided unpaired t-test. k–s, Antibody-induced neutropenia generates atherosclerotic lesions with reduced signs of vulnerability in the brachiocephalic artery in Apoe−/− mice fed a HFD for 16 weeks. k, Representative micrographs showing the aortic arch of isotype IgG (ctrl-IgG) and anti-Ly6G-treated mice stained with H&E (top), Sirius red (middle) and antibodies (bottom) to SMA (red), CD68 (green). l, Experimental scheme. m–s, Analyses of lesion characteristics. Displayed are quantification of intima area (m), fibrous cap (FC) thickness (n), necrotic core area (o), collagen area (p), SMC area (SMA+, q), macrophage area (CD68+, r), and plaque vulnerability (s). n = 8 mice (ctrl IgG), n = 9 mice (anti-Ly6G) except for panel (m) n = 8 mice. Two-sided unpaired t-test. t, u, Proliferating SMCs (Ki67+SMA+) in lesions of indicated treatments. n = 8 mice (ctrl IgG), n = 8 mice (anti-Ly6G), n = 12 mice (PBS), n = 13 mice (AMD3100). Two-sided unpaired t-test. v, w, Percentage of SMC area (SMA+, n = 15 sections for 0.51<, n = 25 sections for >0.51, v) and overall vulnerability (n = 10 sections for 0.51<, n = 20 sections >0.51, w) of human atherosclerotic specimens grouped by the number of lesional neutrophils per mm2. Two-sided unpaired t-test. Data are mean ± s.d. BCA, brachiocephalic artery; ctrl, control; SMA, smooth muscle actin; LCA, left carotid artery; LSA, left subclavian artery; MYH11, Myosin heavy chain 11; RCA, right carotid artery.
