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Nocardia brasiliensis induces formation of foamy macrophages and dendritic cells in vitro and in vivo.

Meester I, Rosas-Taraco AG, Salinas-Carmona MC - PLoS ONE (2014)

Bottom Line: CFSE-labeled BMDM or BMDC were tested for phagocytosis and CD11c/CD11b receptors markers expression before being transferred into the actinomycetoma lesion site of infected mice.Interestingly, many of the transferred BMDM and BMDC were stained with the lipid-droplet fluorophore Nile Red.In conclusion, macrophages and DC cells can be differentiated into foamy cells in vitro and in vivo during N. brasiliensis infection.

View Article: PubMed Central - PubMed

Affiliation: Department of Immunology, Faculty of Medicine, Universidad Autónoma de Nuevo León, Nuevo León, México.

ABSTRACT
Foamy cells have been described in various infectious diseases, for example in actinomycetoma induced by Nocardia brasiliensis. These cells are generally considered to be macrophages, although they present dendritic cell (DC)-specific surface markers. In this study, we determined and confirmed the lineage of possible precursors of foamy cells in vitro and in vivo using an experimental actinomycetoma model in BALB/c mice. Bone marrow-derived macrophages (BMDM) or DC (BMDC) were infected in vitro with N. brasiliensis or labeled with carboxyfluorescein diacetate succinimidyl ester (CFSE). Both, macrophages and DC, differentiated into foamy cells after in vitro infection. CFSE-labeled BMDM or BMDC were tested for phagocytosis and CD11c/CD11b receptors markers expression before being transferred into the actinomycetoma lesion site of infected mice. In vivo studies showed that BMDM and BMDC were traced at the site where foamy cells are present in the experimental actinomycetoma. Interestingly, many of the transferred BMDM and BMDC were stained with the lipid-droplet fluorophore Nile Red. In conclusion, macrophages and DC cells can be differentiated into foamy cells in vitro and in vivo during N. brasiliensis infection.

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Related in: MedlinePlus

Morphology and immunophenotype of bone marrow-derived macrophages (BMDM) and dendritic cells (BMDC).Five-day cultures of BMDM generated with 30% L929-conditioned medium (A) or BMDC generated with 20 ng/mL GM-CSF and IL-4 (B); images were taken at 200x+digital zoom. Over 98% of the BMDM were F4/80+ (C), but less than 4.5% of the macrophage-depleted BMDC were F4/80+ (D).
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pone-0100064-g001: Morphology and immunophenotype of bone marrow-derived macrophages (BMDM) and dendritic cells (BMDC).Five-day cultures of BMDM generated with 30% L929-conditioned medium (A) or BMDC generated with 20 ng/mL GM-CSF and IL-4 (B); images were taken at 200x+digital zoom. Over 98% of the BMDM were F4/80+ (C), but less than 4.5% of the macrophage-depleted BMDC were F4/80+ (D).

Mentions: Murine BMDM are easily generated in vitro with L929-conditioned medium as can be verified by their homogeneous morphology (fig. 1 a) and F4/80 expression in over 98% of the harvested cells (fig. 1c). On the other hand, murine bone marrow cells differentiated with GM-CSF and IL-4 generated a mixture of semi-adherent DC, adherent macrophages, and neutrophils in suspension [25]. The latter were easily removed by elimination of the medium and washing of the adherent cells. However, considering the aim of the study, it was most important to deplete all macrophages from the DC cell cultures. We did not rely on the commonly applied technique of positive selection of CD11c-labeled or CD205-labeled cells because macrophages tend to be CD11c+ [26], [27], and CD205+ macrophages have been described [21], [28]. Therefore, we chose for depletion of F4/80+ cells. F4/80 is considered a macrophage marker, although a subpopulation of DC, for example epidermal Langerhans cells and kidney DC, might also be F4/80+ [29], [30], [31]. To ensure DC (fig. 1b), we preferred losing some DC above having a macrophage contamination. Figure 1d demonstrates that less than 5% of the macrophage-depleted BMDC were F4/80+.


Nocardia brasiliensis induces formation of foamy macrophages and dendritic cells in vitro and in vivo.

Meester I, Rosas-Taraco AG, Salinas-Carmona MC - PLoS ONE (2014)

Morphology and immunophenotype of bone marrow-derived macrophages (BMDM) and dendritic cells (BMDC).Five-day cultures of BMDM generated with 30% L929-conditioned medium (A) or BMDC generated with 20 ng/mL GM-CSF and IL-4 (B); images were taken at 200x+digital zoom. Over 98% of the BMDM were F4/80+ (C), but less than 4.5% of the macrophage-depleted BMDC were F4/80+ (D).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4061056&req=5

pone-0100064-g001: Morphology and immunophenotype of bone marrow-derived macrophages (BMDM) and dendritic cells (BMDC).Five-day cultures of BMDM generated with 30% L929-conditioned medium (A) or BMDC generated with 20 ng/mL GM-CSF and IL-4 (B); images were taken at 200x+digital zoom. Over 98% of the BMDM were F4/80+ (C), but less than 4.5% of the macrophage-depleted BMDC were F4/80+ (D).
Mentions: Murine BMDM are easily generated in vitro with L929-conditioned medium as can be verified by their homogeneous morphology (fig. 1 a) and F4/80 expression in over 98% of the harvested cells (fig. 1c). On the other hand, murine bone marrow cells differentiated with GM-CSF and IL-4 generated a mixture of semi-adherent DC, adherent macrophages, and neutrophils in suspension [25]. The latter were easily removed by elimination of the medium and washing of the adherent cells. However, considering the aim of the study, it was most important to deplete all macrophages from the DC cell cultures. We did not rely on the commonly applied technique of positive selection of CD11c-labeled or CD205-labeled cells because macrophages tend to be CD11c+ [26], [27], and CD205+ macrophages have been described [21], [28]. Therefore, we chose for depletion of F4/80+ cells. F4/80 is considered a macrophage marker, although a subpopulation of DC, for example epidermal Langerhans cells and kidney DC, might also be F4/80+ [29], [30], [31]. To ensure DC (fig. 1b), we preferred losing some DC above having a macrophage contamination. Figure 1d demonstrates that less than 5% of the macrophage-depleted BMDC were F4/80+.

Bottom Line: CFSE-labeled BMDM or BMDC were tested for phagocytosis and CD11c/CD11b receptors markers expression before being transferred into the actinomycetoma lesion site of infected mice.Interestingly, many of the transferred BMDM and BMDC were stained with the lipid-droplet fluorophore Nile Red.In conclusion, macrophages and DC cells can be differentiated into foamy cells in vitro and in vivo during N. brasiliensis infection.

View Article: PubMed Central - PubMed

Affiliation: Department of Immunology, Faculty of Medicine, Universidad Autónoma de Nuevo León, Nuevo León, México.

ABSTRACT
Foamy cells have been described in various infectious diseases, for example in actinomycetoma induced by Nocardia brasiliensis. These cells are generally considered to be macrophages, although they present dendritic cell (DC)-specific surface markers. In this study, we determined and confirmed the lineage of possible precursors of foamy cells in vitro and in vivo using an experimental actinomycetoma model in BALB/c mice. Bone marrow-derived macrophages (BMDM) or DC (BMDC) were infected in vitro with N. brasiliensis or labeled with carboxyfluorescein diacetate succinimidyl ester (CFSE). Both, macrophages and DC, differentiated into foamy cells after in vitro infection. CFSE-labeled BMDM or BMDC were tested for phagocytosis and CD11c/CD11b receptors markers expression before being transferred into the actinomycetoma lesion site of infected mice. In vivo studies showed that BMDM and BMDC were traced at the site where foamy cells are present in the experimental actinomycetoma. Interestingly, many of the transferred BMDM and BMDC were stained with the lipid-droplet fluorophore Nile Red. In conclusion, macrophages and DC cells can be differentiated into foamy cells in vitro and in vivo during N. brasiliensis infection.

Show MeSH
Related in: MedlinePlus