Protocols and Video Articles Authored by Julia Peusquens - JoVE

Fria radikalproducerande myeloid-härledda regulatoriska celler

We show in this communication that although isolated 2018-11-20 · Heterogeneity of Ly6G+ Ly6C+ Myeloid-Derived Suppressor Cell Infiltrates during Staphylococcus aureus Biofilm Infection. Heim CE (1), West SC (2), Ali H (2), Kielian T (3). (1)Department of Pathology and Microbiology, University of Nebraska Medical Center, Omaha, Nebraska, USA. (2)College of Information Science and Technology, University of Differential Induction of Ly6G and Ly6C Positive Myeloid Derived Suppressor Cells in Chronic Kidney and Liver Inflammation and Fibrosis a Representative images of DNA release from Ly6G-positive cells (at 24 h post-Loxo stimulation) resolved by confocal microscopy. b Quantitation of DNA release from Ly6G-positive cells in blood of mice ( n = 4/group) at 24 h after agonist stimulation ( p = 0.016, df = 6). Myeloid-derived suppressor cells (MDSCs) represent a heterogeneous population of CD11b+ cells. According to the surface molecules Ly6G and Ly6C (where Ly6G and Ly6C are lymphocyte antigen 6, locus G and C, respectively), MDSCs are further divided into monocytic (Mo-MDSCs, CD11b+ /Ly6C(high) /Ly6G-) and 2016-10-01 · Whilst positivity for CD11c, Ly6G and NK1.1 discriminates dendritic cells, neutrophils and NK cells respectively, circulating monocytes can be characterized into two populations using cell surface expression of Ly6C.

Ly6g and ly6c positive cells

We did not include the F4/80 marker because it is dispensable when identifying myeloid cells in the mouse (Rose et al., 2012). Differential induction of Ly6G and Ly6C positive myeloid derived suppressor cells in chronic kidney and liver inflammation and fibrosis PLoS One . 2015 Mar 4;10(3):e0119662. doi: 10.1371/journal.pone.0119662. Markers such as CD11b, CD11c, F4/80, Gr-1, Ly6C, and Ly6G have long been used to identify various splenic cell myeloid populations. 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. 2015-03-04 · Ly6C and Ly6G positive myeloid cells accumulate during inflammation and fibrosis in liver and kidney.

Fria radikalproducerande myeloid-härledda regulatoriska celler

Biological context of Ly6g. However, cells expressing the innate markers Gr-1 and CD11c were affected to a greater degree by increasing dose than lymphocytes of the adaptive immune response (Th1, CD4+, CD8+, CD19+), resulting in a change in the balance of innate and adaptive cell numbers to favor innate cells at higher infecting doses . After the indicated times liver, kidney, spleen or bone marrow cells were isolated and analysed by flow cytometry. Histograms depict viable (Hoechst negative), non-parenchymal cells stained with CD11b and Gr-1 (A) or viable, CD11bpos cells stained for Ly6G and Ly6C (B).

Glykolys reglerar expansionen av myeloida härledda

Monocytes typically express Ly6G transiently during development while mature granulocytes and peripheral neutrophils retain expression making Ly6G a good cell surface marker for these populations.

In order to characterise MDSC arising in vivo after chronic inflammation we induced liver fibrosis via bile-duct ligation (BDL) and kidney fibrosis by feeding mice an adenine rich diet . Markers such as CD11b, CD11c, F4/80, Gr‐1, Ly6C, and Ly6G have long been used to identify various splenic cell myeloid populations. 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. This hallmark makes Ly6G a good marker for these particular cell populations.
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Myeloid-derived suppressor cells (MDSCs) represent a heterogeneous population of CD11b+ cells. According to the surface molecules Ly6G and Ly6C (where Ly6G and Ly6C are lymphocyte antigen 6, locus G and C, respectively), MDSCs are further divided into monocytic (Mo-MDSCs, CD11b+ /Ly6C(high) /Ly6G-) and 2016-10-01 · Whilst positivity for CD11c, Ly6G and NK1.1 discriminates dendritic cells, neutrophils and NK cells respectively, circulating monocytes can be characterized into two populations using cell surface expression of Ly6C. OT‐I cells were depleted of Ly6C + cells (histograms, left), then transferred into naïve C57BL/6 mice and 10 days later tetramer (H2K b Tet) positive cells (OT‐I) were isolated from the LNs of recipient mice and analyzed for Ly6C expression. Only cells strongly positive for H2K b Tet were However, when Ly6G + Ly6C + cells were assessed for their levels of CD11b expression, two populations were evident, namely, CD11b high Ly6G + Ly6C + and CD11b low Ly6G + Ly6C + cells . To demonstrate the uniqueness of the CD11b high Ly6G + Ly6C + population in biofilm-infected tissue, we compared staining patterns with those in peripheral blood, which revealed distinct differences ( Fig. 1B ).

S100A9, Arginase 1 and iNOS gene expression in the various CD11b + cell populations was analyzed using Q-PCR.
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Fria radikalproducerande myeloid-härledda regulatoriska celler

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. Ly6c (Left) and Ly6g (Right) overexpression cells were surface stained with ab238132 and anti-HA tag conjugated to Alexa Fluor ® 647. ab238132 does not cross-react with Ly6c. Gated on viable cells. 2017-04-06 · Cells negative for CD3, NK1.1, and CD19 were further analyzed to identify Ly6G + Ly6C int neutrophils and Ly6G lo Ly6C hi CD11b + monocytes.