The macrophage heterogeneity: Difference between mouse peritoneal exudate and splenic F4/80⁺ macrophages

Abstract

Macrophages isolated from various tissues manifest differences in cell shape, the expression of surface markers, as well as metabolic and functional activities. However, the heterogeneity of macrophages expressing the same marker in different tissues has not been fully addressed. In the present study, mouse F4/80+ peritoneal exudate macrophages (PEMs) and splenic macrophages (SPMs) appeared similar in most respects. But the percentages of cells expressing CD80, CD40, MHC-II, TLR2, or TLR4, but not CD11c, CD54, or CD23, in freshly isolated F4/80+ SPMs were significantly higher than those in PEMs, whereas the levels of CD86+ cells in F4/80+ SPMs were markedly lower than those in PEMs. After lipopolysaccharide (LPS) stimulation, F4/80+ SPMs expressed significantly higher levels of CD86, CD40, or MHC-II than F4/80+ PEMs, but not CD11c, CD80, CD54, or CD23. F4/80+ SPMs had remarkably lower non-opsonic phagocytotic capacity against chicken RBCs or allo-T cells than PEMs as determined by two-photon microscopes and flow cytometry. SPMs produced markedly more NO than PEMs when cultured with LPS or allo-T cells. Furthermore, SPMs exhibited stronger immunogenicity than PEMs, as determined by the ability to stimulate T cell proliferation, delayed type hypersensitivity, and IFN-gamma production. The data showed the differences between SPMs and PEMs with regard to the phenotypes, phagocytosis, and immunogenicity, which may offer important information for us to better understand the distinguished immune responses of macrophages in spleens and the peritoneal cavity.

Chemokine-mediated rapid turnover of myeloid-derived suppressor cells in tumor-bearing mice

“…Furthermore, it has been reported that mouse monocytes can differentiate into dendritic cells in the presence of GM-CSF and IL-4, 43 and that monocyte-derived macrophages themselves express DC markers such as CD11c, MHC class II, and CD86 after activation. 44 Together with our results showing that CD11b ϩ Gr-1 ϩ MDSCs in the peripheral tissues of tumor-bearing mice were composed primarily of shortlived, nonproliferating macrophages and neutrophils, it seems more likely that the differentiation of myeloid lineage cells, except for the conventional DCs, were primarily occurred in the BM but not spleen even in the tumor-bearing state.…”

Inflammatory monocytes recruited after skeletal muscle injury switch into antiinflammatory macrophages to support myogenesis

“…Later during muscle regeneration (day 7), CX3CR1 hi /Ly-6C − MO/MP proliferation slowed down simultaneously with the complete diff erentiation into MPs, as shown by their strong F4/80 expression (59). Almost all F4/80 + cells were also positive for CD11c and MHCII, as previously observed after infl ammation in various organs (60,61), and were DEC-205 − . In the blood, Ly-6C − MOs, which come from Ly-6C + cells, are also CD11c + (12).…”

Induction of M2-like macrophages in recipient NOD-scid mice by allogeneic donor CD4+CD25+ regulatory T cells

“…Next, the cells were stained with either 0.25 mg PElabeled anti-IFN-c (GIR-208), IL-4 (11B11), IL-10 (JES5-16E3), IL-12 (C17.8), TNF-a (MP6-XT22) or IL-17A (eBio17B7) mAb for 30 min at room temperature in the dark and washed three times, and 10 000 F4/ 80 1 cells were then analyzed by FCM. 26,28,29 Arginase assay The arginase activity assay was performed as previously described. 30,31 Briefly, the cells were lysed in 0.1% Triton X-100.…”

“…The nitrite concentration was calculated with a sodium nitrite standard curve as reported previously. 28 …”

“…CD4 1 T responders (2310 5 ) and macrophage stimulators (1310 5 ), which were pre-treated with 50 mg/mL mitomycin C, were incubated in triplicate in 0.2 mL medium in Ubottomed 96-well microplates (Costar) at 37 uC in 5% CO 2 . 18,28,34,35 The plates were pulsed with 1 mCi of 3 H-labeled thymidine (radioactivity, 185 GBq/mM; Atomic Energy Research Establishment, Beijing, China) per well on day 3, and after 18 h of further incubation, the cells were harvested onto glass fiber filters with an automatic cell harvester (Tomtec, Toku, Finland). The samples were assayed in a .…”