Dataset Information

Frontline Science: Cytokine-mediated developmental phenotype of mouse eosinophils: IL-5-associated expression of the Ly6G/Gr1 surface Ag.

ABSTRACT: Eosinophils have broad and extensive immunomodulatory capacity; recent studies have focused on the roles of distinct eosinophil subsets in specific tissue microenvironments. Ly6G is a GPI-linked leukocyte surface Ag understood primarily as a marker of mouse neutrophils, although its full function is not known. Here, we show that Ly6G/Gr1, detected by mAbs 1A8 (anti-Ly6G) and RB6-8C5 (anti-Gr1), is detected prominently on a significant fraction of eosinophils from mouse bone marrow and bone marrow-derived culture, with fractions expressing this Ag increasing in IL-5-enriched microenvironments. Among our findings, we identified SiglecF⁺ Gr1⁺ eosinophils in bone marrow from naïve, allergen-challenged and IL-5 transgenic mice; SiglecF⁺ Gr1⁺ eosinophils were also prominent ex vivo in bone marrow-derived eosinophils (bmEos) in IL-5-enriched culture. Reducing the IL-5 concentration 20-fold had no impact on the rate of generation of SiglecF⁺ bmEos but did result in a marked increase in the Gr1^- fraction (from 17.4 ± 2% to 30 ± 2.3%, ***P < 0.005). Reducing the IL-5 concentration also enhanced chemotaxis; SiglecF⁺ Gr1^- bmEos were considerably more responsive to eotaxin-1 than were their SiglecF⁺ Gr1⁺ counterparts. These results suggest that (i) IL-5 regulates the expression of Ly6G/Gr1, either directly or indirectly, in cells of the eosinophil lineage, (ii) eosinophils generated in response to high concentrations of IL-5 can be distinguished from those generated under homeostatic conditions by expression of the Ly6G/Gr1 cell surface Ag, and (iii) expression of Ly6G/Gr1 may have an impact on function, directly or indirectly, including the potential to undergo chemotaxis in response to eotaxin-1.

SUBMITTER: Limkar AR

PROVIDER: S-EPMC7044020 | biostudies-literature | 2020 Mar

REPOSITORIES: biostudies-literature

ACCESS DATA

Publications

Frontline Science: Cytokine-mediated developmental phenotype of mouse eosinophils: IL-5-associated expression of the Ly6G/Gr1 surface Ag.

Limkar Ajinkya R AR Mai Eric E Sek Albert C AC Percopo Caroline M CM Rosenberg Helene F HF

Journal of leukocyte biology 20191101 3

Eosinophils have broad and extensive immunomodulatory capacity; recent studies have focused on the roles of distinct eosinophil subsets in specific tissue microenvironments. Ly6G is a GPI-linked leukocyte surface Ag understood primarily as a marker of mouse neutrophils, although its full function is not known. Here, we show that Ly6G/Gr1, detected by mAbs 1A8 (anti-Ly6G) and RB6-8C5 (anti-Gr1), is detected prominently on a significant fraction of eosinophils from mouse bone marrow and bone marro ...[more]

PMID: 31674692

Similar Datasets

Project description:Bone marrow is a hematopoietic site harboring multiple populations of myeloid cells in different stages of differentiation. Murine bone marrow eosinophils are traditionally identified by Siglec-F(+) staining using flow cytometry, whereas neutrophils are characterized by Ly6G(+) expression. However, using flow cytometry to characterize bone marrow hematopoietic cells in wild-type mice, we found substantial gray areas in identification of these cells. Siglec-F(+) mature eosinophil population constituted only a minority of bone marrow Lin(+)CD45(+) pool (5%). A substantial population of Siglec-F(-) cells was double positive for neutrophil marker Ly6G and eosinophil lineage marker, IL-5Rα. This granulocyte population with mixed neutrophil and eosinophil characteristics is typically attributable to neutrophil pool based on neutral granule staining and expression of Ly6G and myeloid peroxidase. It is distinct from Lineage(-) myeloid progenitors or Siglec-F(+)Ly6G(+) maturing eosinophil precursors, and can be accurately identified by Lineage(+) staining and positive expression of markers IL-5Rα and Ly6G. At 15-50% of all CD45(+) hematopoietic cells in adult mice (percentage varies by sex and age), this is a surprisingly dominant population, which increases with age in both male and female mice. RNA-seq characterization of these cells revealed a complex immune profile and the capacity to secrete constituents of the extracellular matrix. When sorted from bone marrow, these resident cells had neutrophilic phenotype but readily acquired all characteristics of eosinophils when cultured with G-CSF or IL-5, including expression of Siglec-F and granular proteins (Epx, Mbp). Surprisingly, these cells were also able to differentiate into Ly6C(+) monocytes when cultured with M-CSF. Herein described is the discovery of an unexpected hematopoietic flexibility of a dominant population of multipotent myeloid cells, typically categorized as neutrophils, but with the previously unknown plasticity to contribute to mature pools of eosinophils and monocytes.

Dataset Information

Frontline Science: Cytokine-mediated developmental phenotype of mouse eosinophils: IL-5-associated expression of the Ly6G/Gr1 surface Ag.

Publications

Frontline Science: Cytokine-mediated developmental phenotype of mouse eosinophils: IL-5-associated expression of the Ly6G/Gr1 surface Ag.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets