Fig 2: The effects of granulocyte depletion on BsAb directed T cell trafficking and in vivo anti-tumor response. A Luciferase transduced T cells [Luc(+) T cells] were administered with GD2-BsAb and anti-GR1 or anti-Ly6G antibody to treat small cell lung cancer cell line (NCI-N417) xenografts. Luc(+) T cells were administered intravenously (iv) on day 0, and GD2-BsAbs were iv administered on day 0 and day 5 separately. Anti-Ly6G or anti-GR1 antibodies were administered on day -1 and day 4 intraperitoneally. B Bioluminescence of Luc(+) T cells infiltrated into tumors were monitored. The bioluminescence images on day 4 and quantification of the bioluminescence in the lesions of tumor. Control BsAb, Luc(+) T cells plus control BsAb; GD2-BsAb, Luc(+) T cells plus GD2-BsAb; GD2-BsAb plus anti-GR1, Luc(+) T cells plus GD2-BsAb plus anti-GR1 antibody; GD2-BsAb plus anti-Ly6G, Luc(+) T cells plus GD2-BsAb plus anti-Ly6G antibody. C In vivo anti-tumor response by GD2-BsAb and granulocyte depleting antibodies was tested against melanoma cell line (M14) xenografts. Control BsAb, Luc(+) T cells plus control BsAb; GD2-BsAb, Luc(+) T cells plus GD2-BsAb; GD2-BsAb+ anti-GR1, Luc(+) T cells plus GD2-BsAb plus anti-GR1 antibody; GD2-BsAb+ anti-Ly6G, Luc(+) T cells plus GD2-BsAb plus anti-Ly6G antibody. D In vivo anti-tumor effect of GD2-EATs with anti-Ly6G antibody was tested against neuroblastoma patient-derived xenografts (PDXs), and long-term survival was analyzed

Fig 3: Limited accumulation of myeloid cells in AAA of IL-27R-deficient mice. Single-cell suspensions of suprarenal aortas/AAA of Apoe-/-Il27ra+/- (n = 6) or Apoe-/-Il27ra-/- (n = 8) mice fed with WD and infused with Ang II were stained for Live/Dead, CD45, CD11b, CD11c, Ly6G, Ly6C, and analyzed by FACS. a Left: representative FACS plots of CD45+ cells. Numbers indicate percentage of cells in each gate. a Right: absolute number of live CD45+ cells. b Representative FACS plots and absolute number of CD45+ live CD11b+, CD11b+CD11c+, and CD11c+ cells. c Representative FACS plots and absolute number of live CD11b+Ly6Chi, CD11b+Ly6G+ (Neutrophils), and CD11b+Ly6Clow cells. **p < 0.01, ***p < 0.005, unpaired Student’s t-test (two-tailed). d Immunofluorescent confocal analysis of localization and abundance of CD11b+ and Ly6G+ (Neutrophils) cells in suprarenal aortas/AAA of Apoe-/-Il27ra+/- and Apoe-/-Il27ra-/- mice infused with Ang II. L-lumen, M-media, A-adventitia, AAA-abdominal aortic aneurysm. Representative images from three independent experiments. Relative gene expression of chemokines (e), cytokines (f), and MMPs (g) in suprarenal aortas/AAA of Apoe-/-Il27ra-/- mice were normalized to Rpl32 expression and then normalized to gene expression in control Apoe-/-Il27ra+/- mice. **p < 0.01, ***p < 0.005, Wilcoxon’s signed-rank test. h Leukocytes, monocytes, and neutrophils counts were evaluated by VetScan HM5 in the blood of Apoe-/-Il27ra+/- (n = 5) or Apoe-/-Il27ra-/- (n = 5) mice prior to and on day 5 after pump implantation. Data are mean ± SEM from at least three independent experiments. *p < 0.05. **p < 0.01, ***p < 0.005, unpaired Student’s t-test (two-tailed)

Fig 4: JHQG attenuates neutrophil accumulation in the lungs of LPS-induced ALI mice. (A) Neutrophil accumulation for lung tissues were detected by immunofluorescent staining for the neutrophil marker Ly6G (n = 3) (original magnification of whole field, 200 ×; enlarged box magnification, 630×). Ly6G: red; DAPI: blue. (B) The number of neutrophils stained for Ly6G per section are presented as the mean ± SD. (C) Total cells in BALF were determined using a Cell Counter (n = 6). MPO activity in lung tissues (D) and MIP-2 level in BALF (E) were measured at 24 h after LPS stimulation (n = 6). Data are expressed as the mean ± SD. Comparison to CON group, #p < 0.05; comparison to LPS group, *p < 0.05; One-way ANOVA. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Fig 5: IFN-? regulates macrophage inflammatory profile during normal tissue repair.(A) IFN-?–KO and control wound cell isolates were processed for flow cytometry using the following gating strategy selecting for single cells, live (viability dye–), CD45+, Ly6G-, CD11b+, Ly6Chi, or Ly6Clo. (B and C) Flow cytometry quantification of Ly6Chi and Ly6Clo cells in wounds (n = 4–6 per group). (D) Wound monocyte/macrophages (MF) (CD3–CD19–Ly6G–CD11b+) were isolated from WT and IFN-?–KO mice on day 3; n = 3 per group, repeated in triplicate. Gene expression of inflammatory cytokines Tnf, Il1b, and Il12 was measured via qPCR. Data were analyzed for variances, and 2-tailed Student’s t test or 1-way ANOVA was performed. *P < 0.05, **P < 0.01, and ***P < 0.001. Data are presented as mean ± SEM.