12 Dec 2012 CD11b. +. Ly6C. ++. Ly6G. - cells show distinct function in mice with chronic inflammation or tumor burden. Eva Källberg, Martin Stenström,
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. 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 Methods: CD11b(+)Ly6C(++) and Ly6G(+) cells were isolated from spleen, tumor tissue or inflammatory granulomas. S100A9, Arginase 1 and iNOS gene expression in the various CD11b(+) cell populations was analyzed using Q-PCR. The suppressive activity of the CD11b(+) cell populations from different donors was studied in co-culture experiments.
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Anti-F4/80. MicroBeads. UltraPure. CD8+ Dendritic Cell Isolation Kit Ly6C+/–. Siglec-F+/–. Ly-6G+/–. CD11c+/–.
Ly-6G+/–. Siglec-F+/– CD11b–. Ly -6Chigh.
26 May 2020 CD11bhiLy6ChiLy6Glo cells were isolated from BM cells cultured for 5 days under GM-CSF incubation (40 ng/mL) but without MSC coculture.
CD11b + /Ly6C med /Ly6G +, but not CD11b + /Ly6C high /Ly6G − cells, were reduced when MCP-1 was neutralized in HCM but not NCM-treated mice (Supplementary Fig. S3A and B). Thus, accumulation of granulocytic CD11b + /Ly6C med /Ly6G + myeloid cells, driven by hypoxic tumor cell–derived MCP-1, regulates the prometastatic properties of the premetastatic niche. 2018-11-20 · CD11bhigh Ly6G+ Ly6C+ MDSCs suppressed T cell proliferation throughout the 28-day infection period, whereas CD11blow Ly6G+ Ly6C+ PMNs had no effect early (day 3 postinfection), although this population acquired suppressive activity at later stages of biofilm development.
Cd11b+Gr1+ cells and identify subpopulations accumulating in the premetastatic lungs (Fig. 1D). Because the anti-Gr1 antibody Ly6G-Ly6C- cells, suggesting that the primary tumor secretes fac-tors that speciﬁcally up-regulate Bv8 expression in Ly6G+Ly6C+ granulocytes
The suppressive activity of the CD11b(+) cell populations from different donors was studied in co-culture experiments. The myeloid differentiation antigen Gr-1 consists of two epitopes, recognized by anti-lymphocyte antigen (Ly) 6G and anti-Ly6C antibodies, which divide CD11b + Gr-1 + MDSCs into Ly6G + granulocytes and Ly6C + monocytes . These two subpopulations may have different functions in infectious diseases and cancer (34–36). 2017-07-10 · Previous studies have focused on Gr1 hi (Ly6C hi or Ly6G +) myeloid cells (26, 28, 35 – 43). In this study, we clearly discriminate between Ly6C hi and Ly6G + myeloid cell subsets based on their immunophenotype (i.e., Ly6C hi monocytes and Ly6G + neutrophils, respectively) (gated on CD45 + Lin – F4/80 – CD11c – CD11b +).
Most studies have focused on either CD11b+Ly6G+Ly6C+ granulocytic or polymorphonuclear myeloid-derived suppressor cells (G-MDSCs or PMN-MDSCs) or CD11b+Ly6GLy6C+ monocytic MDSCs (M-MDSCs), for which clear roles have been established. On the other hand, CD11b+Ly6GLy6C myeloid-derived cells (MDCs) have been less well studied. CD11b + Ly6C ++ and Ly6G + cells were isolated from spleen, tumor tissue or inflammatory granulomas. S100A9, Arginase 1 and iNOS gene expression in the various CD11b + cell populations was analyzed using Q-PCR.
The RB6-8C5 monoclonal antibody reacts strongly with mouse Ly6G and weakly with mouse Ly6C previously referred to as GR-1.
(B) CD11b + Ly6G − myeloid cells can be separated into three populations based on Ly6C expression, with Ly6C low myeloid cells constituting the bulk of these cells in the naïve animal. CD11b + Ly6G − Ly6C low cells show a biphasic response after CFA injection, peaking at 24 h and again at 14 d, whereas they make up the majority of cells between 3 and 10 d after plantar incision.
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Ly6G (Lymphocyte antigen 6 complex locus G6D) is a 21-25kD glycosylphosphatidylinositol (GPI)-linked differentiation antigen that is expressed by myeloid-derived cells in a tightly developmentally-regulated manner in the bone marrow. Monocytes express Ly6G transiently during bone marrow development, while Ly6G expression in granulocytes and peripheral neutrophils directly correlates with the
The efficiency of the depletion needs to be carefully analysed as flow cytometry plots may be misinterpreted. For example, the staining intensity of GR1 on neutrophils (CD11b + GR1 hi) drops upon anti-Ly6G administration Colorectal immunocytes were stained with anti-CD11b+ Ly6C+ (101206, 128008, Biolegend) to determine monocytic myeloid-derived suppressor cells (M-MDSCs), anti-CD11b+ Ly6G+ (101206, 1127627, Biolegend) to granulocytic myeloid-derived suppressor cells (G-MDSC).