(D) Representative FACS plots of CD11b+,Ly6G+Ly6C- neutrophils, Ly6C high Ly6G- monocytes, and F4/80+ macrophages in the blood of CVB3-infected mice treated with anti-Ly6G or anti-Gr1 on day7 p.i. (pre

2017-10-31 · Interestingly, resident Ly6C-Ly6G-CD11b+ macrophages were also infected, however they represented the least infected cells among the three populations (Fig. 1C–F).

Neutrophils (R1, Gr‐1 high F4/80 neg cells) express Ly6C and Ly6G; macrophages (R2, Gr‐1 int F4/80 + cells) express Ly6C but not Ly6G. Dotted line: Isotype control antibody. Solid line: Anti‐Ly6C or anti‐Ly6G mAb. The Ly6C+Ly6G- (top, open histograms) and Ly6C+Ly6G+ (bottom, open histograms) CD11b+ monocyte subpopulations were analyzed for the expression of various cytokines/effector molecules using antibodies (A-D) or a reactive dye triggered by exposure to ROS (E) and compared to CD11b- cells (filled histograms).

Most studies have focused on either CD11b+Ly6G+Ly6C+ granulocytic or polymorphonuclear myeloid-derived suppressor cells (G-MDSCs or PMN-MDSCs) or CD11b+Ly6G−Ly6C+ monocytic MDSCs (M-MDSCs), for which clear roles have been established. Ly6C hi monocytes digest damaged tissue, whereas Ly6C lo monocytes promote healing via myofibroblast accumulation, angiogenesis and deposition of collagen . It appears that Ly6C hi monocytes cooperate with M1 macrophages in inflammatory functions, whereas Ly6C lo monocytes work together with M2 macrophages to achieve angiogenic functions . 2014-03-13 Further staining with Ly6C and Ly6G revealed three populations of “L-DC candidates” with overlapping FSC and SSC profiles: Ly6C lo Ly6G + , Ly6C lo Ly6G − and Ly6C − Ly6G − . Gating strategies were based on fluorescence minus one control, and numbers in gates represent % specific binding. 2020-03-04 cells Article Distinct Role of CD11b+Ly6G Ly6C Myeloid-Derived Cells on the Progression of the Primary Tumor and Therapy-Associated Recurrent Brain Tumor Sheng-Yan Wu 1 and Chi-Shiun Chiang 1,2,3,* 1 Department of Biomedical Engineering and Environmental Sciences, National Tsing Hua University, 101 Sec. 2, Kuang-Fu Road, Hsinchu 30013, Taiwan; z2743216810@hotmail.com.tw 2019-12-24 2008-04-01 Purified CD11b + Ly6C hi Ly6G − and CD11b + Ly6C int Ly6G + cells were seeded into 24-well tissue culture plates at 1 × 10 5 cells per well using medium formulated for the ex vivo culture of bone marrow-derived macrophages (Dulbecco's modified Eagle medium [DMEM] with GlutaMAX-I [Invitrogen] supplemented with 20% heat-inactivated fetal bovine serum [Atlanta Biologicals], 20% L-cell (B) Colonic cell population from IELs and LPMCs of untreated WT mice (H 2 O) and DSS-treated WT and gp130 757F/F mice were characterized by flow cytometry for subpopulations of CD11c + F4/80 − Ly6G − Ly6C − dendritic cells (DC), CD11c − CD11b + Ly6G − F4/80 + monocytes (Mo), CD11b − F4/80 + Ly6C − Ly6G − macrophages (MΦ) and CD11b + Ly6G + Ly6C low F4/80 − CD11c 2011-09-01 2020-05-05 Inflammatory mediators can activate and sensitize nociceptors, specialized high-threshold nerve fibers that relay noxious signals to the spinal cord and brain to initiate pain. However, the contribution of specific immune cell types to pain in animal models of inflammation remains largely unknown.

1C–F).

(B) Colonic cell population from IELs and LPMCs of untreated WT mice (H 2 O) and DSS-treated WT and gp130 757F/F mice were characterized by flow cytometry for subpopulations of CD11c + F4/80 − Ly6G − Ly6C − dendritic cells (DC), CD11c − CD11b + Ly6G − F4/80 + monocytes (Mo), CD11b − F4/80 + Ly6C − Ly6G − macrophages (MΦ) and CD11b + Ly6G + Ly6C low F4/80 − CD11c

Eosinophils and monocytes are negative for this marker. Neutrophils: CD11c- CD11b+ Ly6G+ , Monocytes: CD11b+ CD11c- Ly6c + - ly6G -.

CD11b+Ly6C high (Ly6Chi) cells (inflammatory monocytes) and CD11b+Ly6C low cells (reparative monocytes) were recruited into the liver following I/R injury. Treatment with clodronate‑loaded liposomes, which transiently deplete systemic macrophages, alleviated hepatic damage.

Collectively, these data identify kinetic and functional differences in diabetic wound monocyte/macrophages and demonstrate that selective targeting of CD11b⁺Ly6C^Hi monocyte/macrophages is a viable therapeutic strategy for inflammation in diabetic wounds. Because Ly6C Lo [Lin − Ly6G − CD11b +] monocyte/macrophages represent more of a hybrid picture between in vitro M1 and M2 macrophages, it is possible that the use of traditional in vitro macrophage characterization for this cell population in wound healing is not practical, as has been previously suggested. 21 These findings are highly relevant as they will allow for more accurate immunophenotyping of wound monocyte/macrophages and identify new avenues for therapy. Ly6G is a glycosylphosphatidylinositol‐linked cell‐surface protein that is expressed predominantly on murine neutrophils .

In skin, the inflammatory Mo/MΦ population was defined as Live Ly6G-CD11b+Ly6C+F4/80lo/-, while mature MΦ were gated as Live Ly6G-CD11b+ Ly6C- F4/80+. Mo were defined as Ly6G-cKit-CD11b+CD115+Ly6Chi/lo in bone marrow and Ly6G-CD11b+CD115+Ly6Chi/lo in peripheral blood. The Ly6C Int Ly6G-SSC Hi population consisted almost entirely of eosinophils with very rare monocytes/macrophages (Figures 3a and 3b, Gate B). Ly6C Lo-neg Ly6G-SSCLo and Ly6C + Ly6G-SSC Lo cells were all monocytes and macrophages.
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Ear pinnae were harvested at 5 days post-infection. The Ly6C+Ly6G- (top, open histograms) and Ly6C+Ly6G+ (bottom, open histograms) CD11b+ monocyte subpopulations were analyzed for the expression of various cytokines/effector molecules using … 2017-10-31 2019-03-29 2010-07-15 2019-03-29 2017-12-29 We sequenced RNA in mouse CD11b + Ly6G − Ly6C high and CD11b + Ly6G − Ly6C low monocytes sorted from WT whole blood. Sorted cells (200 000/monocyte subset) were collected in 1400 μL QIAzol Lysis Reagent (Qiagen, Germantown, MD) and total RNA (50–100 ng) was isolated manually per manufacturer’s protocol.

Using flow-sorted tdTomato -labeled Ly6C Hi monocyte/macrophages, we show Ly6C Hi cells transition to a Ly6C Lo phenotype in normal wounds, whereas in diabetic wounds, there is a late, second influx of Ly6C Hi cells that fail transition to Ly6C Lo. Flow cytometry and fluorescence‐activated cell sorting (FACS) analysis demonstrated that Ly6G/Ly6C markers are superior to Gr‐1 for identifying splenic neutrophils, eosinophils, and subsets of monocytes/macrophages. Ly6G is a glycosylphosphatidylinositol‐linked cell‐surface protein that is expressed predominantly on murine neutrophils [ 1 ]. The function of Ly6G has still not been fully clarified, although a recent study implicates Ly6G in neutrophil migration [ 2 ]. There are 2 mAb, i.e., RB6‐8C5 and 1A8, which recognize Ly6G [ 3 ].
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Because Ly6C Lo [Lin − Ly6G − CD11b +] monocyte/macrophages represent more of a hybrid picture between in vitro M1 and M2 macrophages, it is possible that the use of traditional in vitro macrophage characterization for this cell population in wound healing is not practical, as has been previously suggested. 21 These findings are highly relevant as they will allow for more accurate immunophenotyping of wound monocyte/macrophages and identify new avenues for therapy.

Histograms: Wound cells were stained with Gr‐1, F4/80, and Ly6C or Ly6G. Neutrophils (R1, Gr‐1 high F4/80 neg cells) express Ly6C and Ly6G; macrophages (R2, Gr‐1 int F4/80 + cells) express Ly6C but not Ly6G. Dotted line: Isotype control antibody. Solid line: Anti‐Ly6C or anti‐Ly6G mAb. both Ly6G and Ly6C), monocytes (expressing Ly6C but not Ly6G), macrophages, dendritic cells, and myeloid suppressor cells (20, 22). We observed a marked enrichment (ﬁve- to six-fold) in Ly6C+Ly6G+ granulocytes and a modest accumulation of Ly6C+Ly6G- monocytes and F4/80+ macrophages in the premetastatic lungs of 4T1 tumor-bearing mice (Fig. 1D Ly6C hi MHC II − cells (gate 1: Fig. 1A) were F4/80 + CX 3 CR1 low CCR2 hi CD62L +, did not express the granulocyte markers Ly6G or CCR3, and had a small size and granularity (FSC low SSC low), indicating that they were Ly6C hi monocytes (Fig.