Immature macrophages derived from mouse bone marrow produce large amounts of IL-12p40 after LPS stimulation

Oliveira, Milton Adriano Pelli de; Lima, Guilherme Meira; Shio, Marina Tiemi; Leenen, Pieter J. M.; Abrahamsohn, Ises A.

doi:10.1189/jlb0302124

Cited by 23 publications

(10 citation statements)

References 47 publications

(34 reference statements)

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“…It has been reported that enteric bacteria play essential roles in onset and development of colitis in IL-10 Ϫ/Ϫ mice, similar to human IBDs (15) In the present study, only stimuli from whole bacteria, but not from PAMPs could induce the production of IL-12p70, IL-12 bioactive form consisted of p35-p40 heterodimer, in differentiated M from BM CD11b ϩ cells. In general, TLR ligands such as LPS only induce IL-12p40 subunits, but fail to induce IL-12p70 production from M without IFN-␥ costimulation (36,37). Consistent with this, we also demonstrated that GM-M and M-M produced none or just low levels of IL-12p70 in response to stimulation with various TLR ligands.…”

Section: Discussionsupporting

confidence: 74%

Abnormally Differentiated Subsets of Intestinal Macrophage Play a Key Role in Th1-Dominant Chronic Colitis through Excess Production of IL-12 and IL-23 in Response to Bacteria

Kamada

Hisamatsu

Okamoto

et al. 2005

The Journal of Immunology

192

185

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Disorders in enteric bacteria recognition by intestinal macrophages (Mφ) are strongly correlated with the pathogenesis of chronic colitis; however the precise mechanisms remain unclear. The aim of the current study was to elucidate the roles of Mφ in intestinal inflammation by using an IL-10-deficient (IL-10−/−) mouse colitis model. GM-CSF-induced bone marrow-derived Mφ (GM-Mφ) and M-CSF-induced bone marrow-derived Mφ (M-Mφ) were generated from bone marrow CD11b+ cells. M-Mφ from IL-10−/− mice produced abnormally large amounts of IL-12 and IL-23 upon stimulation with heat-killed whole bacteria Ags, whereas M-Mφ from wild-type (WT) mice produced large amounts of IL-10 but not IL-12 or IL-23. In contrast, IL-12 production by GM-Mφ was not significantly different between WT and IL-10−/− mice. In ex vivo experiments, cytokine production ability of colonic lamina propria Mφ (CLPMφ) but not splenic Mφ from WT mice was similar to that of M-Mφ, and CLPMφ but not splenic Mφ from IL-10−/− mice also showed abnormal IL-12p70 hyperproduction upon stimulation with bacteria. Surprisingly, the abnormal IL-12p70 hyperproduction from M-Mφ from IL-10−/− mice was improved by IL-10 supplementation during the differentiation process. These results suggest that CLPMφ and M-Mφ act as anti-inflammatory Mφ and suppress excess inflammation induced by bacteria in WT mice. In IL-10−/− mice, however, such Mφ subsets differentiated into an abnormal phenotype under an IL-10-deficient environment, and bacteria recognition by abnormally differentiated subsets of intestinal Mφ may lead to Th1-dominant colitis via IL-12 and IL-23 hyperproduction. Our data provide new insights into the intestinal Mφ to gut flora relationship in the development of colitis in IL-10−/− mice.

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Section: Discussionsupporting

confidence: 74%

Abnormally Differentiated Subsets of Intestinal Macrophage Play a Key Role in Th1-Dominant Chronic Colitis through Excess Production of IL-12 and IL-23 in Response to Bacteria

Kamada

Hisamatsu

Okamoto

et al. 2005

The Journal of Immunology

192

185

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“…The result showed that after 72 hours exposure to 0.5 μ g/mL of rSj16, the nuclear: cytoplasmic ratio of the cells reduced, the nuclear chromatin became condensed and eccentrically placed, the cytoplasmic borders were protruded, and cytoplasmic vacuoles were often present (Figure 3(e)). These morphological features were similar to those described for mature tissue macrophages [15–17]. WEHI-3B JCS cells treated with rGST did not exhibit significant morphological changes (Figure 3(d)).…”

Section: Resultssupporting

confidence: 85%

Anti-Inflammatory Protein ofSchistosoma japonicumDirects the Differentiation of the WEHI-3B JCS Cells and Mouse Bone Marrow Cells to Macrophages

Mak

et al. 2010

Journal of Biomedicine and Biotechnology

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Sj16 is an anti-inflammatory protein identified from Schistosoma japonicum. Our previous studies showed that recombinant Sj16 (rSj16) could suppress host's inflammatory responses and inhibit macrophage maturation. In the present study, the effects of rSj16 on the differentiation of the murine myeloid leukemia WEHI-3B JCS cell line and on mouse hematopoiesis were investigated. Our data demonstrated that rSj16 expressed and purified from Escherichia coli could suppress the proliferation of the WEHI-3B JCS cells in a time- and concentration-dependent manner, while not affect the viability of the cells. Further studies indicated that rSj16 induced macrophage differentiation of the WEHI-3B JCS cells, and arrested the cell cycle in the G1/G0 and G2/M phases. The macrophage differentiation of the rSj16-treated WEHI-3B JCS cells was confirmed by their expression of macrophage specific antigen F4/80 and phagocytic activity. Furthermore, our results revealed that rSj16 biased the colony formation of mouse bone marrow cells towards macrophage linage.

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“…One way to test the effect of stromal cells in BMDM maturation would be to co-culture bone marrow stromal cell line such as M2-10B4 with the myeloid progenitor cells at different ratios and characterizing the final BMDM population. In regards to the relationship between the maturation states of a monocytic cell to their polarization propensity, an earlier study had indicated that immature macrophages secrete more IL-12 [9]. However, a maturation status of a macrophage might be more complex than just defined by cell surface markers or morphology.…”

Section: Resultsmentioning

confidence: 99%

Cell density during differentiation can alter the phenotype of bone marrow-derived macrophages

Lee

2013

Cell Biosci

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BackgroundBone marrow-derived macrophages (BMDMs) are widely used primary cells for studying macrophage function. However, despite numerous protocols that are currently available, lack of a notable consensus on generating BMDMs may obscure the reliability in comparing findings from different studies or laboratories.FindingsIn this study, we addressed the effect of cell density on the resulting macrophage population. With reference to previously published methods, bone marrow cells from wild type C57BL/6 mice were plated at either 4 × 105 cells or 5 × 106 cells per 10 cm and cultured in 20% L-cell conditioned media for 7 days, after which they were analyzed for cell surface markers, production of proinflammatory cytokines, and responsiveness to polarizing signals. Reproducibly, cells plated at lower density gave a pure population of CD11b+F4/80+ macrophages (97.28 ± 0.52%) with majority being Ly-6C-Ly-6G- and c-Fms+, while those plated at higher density produced less CD11b+F4/80+ cells and a considerably higher proportion of CD11b+F4/80+CD11c+ (68.72 ± 2.52%) and Ly-6C-Ly-6G+ (71.10 ± 0.90%) cells. BMDMs derived from higher plating density also secreted less proinflammatory cytokines such as IL-6, IL-12 and TNF-α and were less phagocytic, and had a different pattern of expression for M1- and M2-related genes upon LPS or IL-4 stimulation.ConclusionsOverall, our findings indicate that altering cell density during BMDM differentiation can give rise to distinct macrophage populations that could vary the outcome of a functional study.

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Immature macrophages derived from mouse bone marrow produce large amounts of IL-12p40 after LPS stimulation

Cited by 23 publications

References 47 publications

Abnormally Differentiated Subsets of Intestinal Macrophage Play a Key Role in Th1-Dominant Chronic Colitis through Excess Production of IL-12 and IL-23 in Response to Bacteria

Abnormally Differentiated Subsets of Intestinal Macrophage Play a Key Role in Th1-Dominant Chronic Colitis through Excess Production of IL-12 and IL-23 in Response to Bacteria

Anti-Inflammatory Protein ofSchistosoma japonicumDirects the Differentiation of the WEHI-3B JCS Cells and Mouse Bone Marrow Cells to Macrophages

Cell density during differentiation can alter the phenotype of bone marrow-derived macrophages

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