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Rat macrophage C-type lectin is an activating receptor expressed by phagocytic cells.

Lobato-Pascual A, Saether PC, Dahle MK, Gaustad P, Dissen E, Fossum S, Daws MR - PLoS ONE (2013)

Bottom Line: We found that MCL co-immunoprecipitated with the activating adaptor protein FcεRIγ in these cells.Moreover, beads coated with anti-MCL antibody increased phagocytosis in the RMW cells.Together, these observations indicate that rat MCL is a receptor that activates phagocytosis in myeloid cells under inflammatory conditions.

View Article: PubMed Central - PubMed

Affiliation: Department of Anatomy, Institute of Basic Medical Sciences, University of Oslo, Oslo, Norway. a.l.pascual@medisin.uio.no

ABSTRACT
Macrophage C-type lectin (MCL) is a membrane surface receptor encoded by the Antigen Presenting Lectin-like gene Complex (APLEC). We generated a mouse monoclonal antibody for the study of this receptor in the rat. We demonstrate that rat MCL is expressed on blood monocytes and neutrophils, as well as on several tissue macrophage populations, including alveolar and peritoneal cavity macrophages. We also demonstrate MCL expression on a subset of resident spleen macrophages. Immunohistochemistry analysis of the spleen showed staining specifically in the marginal zone and red pulp. Exposure to pro-inflammatory mediators or to yeast cell wall extract (zymosan) increased surface MCL expression on peritoneal macrophages. We characterized a rat myeloid cell line, RMW, which expresses high levels of MCL. We found that MCL co-immunoprecipitated with the activating adaptor protein FcεRIγ in these cells. Moreover, beads coated with anti-MCL antibody increased phagocytosis in the RMW cells. Together, these observations indicate that rat MCL is a receptor that activates phagocytosis in myeloid cells under inflammatory conditions.

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Regulation of MCL surface expression under the effect of different stimuli.A, Rats were injected intraperitoneally with zymosan, peritoneal cells were obtained after 24 h and stained with mAbs for flow cytometry analysis. Discontinuous line histogram shows staining in the zymosan treated group, dotted line histogram shows MCL staining in the PBS control group. Histograms display MCL expression on granular cells (including mast cells, eosinophils and neutrophils) and macrophages (MΦ) as indicated. The MCL+ fraction of granular cells likely reflects influx of neutrophils from the blood. MFI values are shown, calculated as the median of the fluorescence intensity. B, Resident peritoneal macrophages were cultured overnight with the indicated substances. Solid line histogram shows MCL staining. Grey filled histograms correspond to isotype controls.
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pone-0057406-g003: Regulation of MCL surface expression under the effect of different stimuli.A, Rats were injected intraperitoneally with zymosan, peritoneal cells were obtained after 24 h and stained with mAbs for flow cytometry analysis. Discontinuous line histogram shows staining in the zymosan treated group, dotted line histogram shows MCL staining in the PBS control group. Histograms display MCL expression on granular cells (including mast cells, eosinophils and neutrophils) and macrophages (MΦ) as indicated. The MCL+ fraction of granular cells likely reflects influx of neutrophils from the blood. MFI values are shown, calculated as the median of the fluorescence intensity. B, Resident peritoneal macrophages were cultured overnight with the indicated substances. Solid line histogram shows MCL staining. Grey filled histograms correspond to isotype controls.

Mentions: Receptor expression under inflammatory conditions was examined by inducing sterile peritonitis with injections of zymosan in the peritoneal cavity of rats. 24 h after treatment, cells were collected by lavage of the peritoneal cavity and analyzed by flow cytometry. In the zymosan treated animals, MCL positive granulocytes appeared, corresponding to an expected influx of neutrophils. MCL expression was in addition increased on macrophages (Figure 3A). These were isolated from the peritoneum, maintained in vitro either in M-CSF, IFNγ plus LPS, or in IL-4. MCL was clearly increased when exposed to IFNγ and pro-inflammatory stimuli from G– bacteria, but it decreased following culture in the presence of anti-inflammatory IL-4 (Figure 3B). Taken together, these data suggest that MCL plays a role in inflammatory responses to microorganisms.


Rat macrophage C-type lectin is an activating receptor expressed by phagocytic cells.

Lobato-Pascual A, Saether PC, Dahle MK, Gaustad P, Dissen E, Fossum S, Daws MR - PLoS ONE (2013)

Regulation of MCL surface expression under the effect of different stimuli.A, Rats were injected intraperitoneally with zymosan, peritoneal cells were obtained after 24 h and stained with mAbs for flow cytometry analysis. Discontinuous line histogram shows staining in the zymosan treated group, dotted line histogram shows MCL staining in the PBS control group. Histograms display MCL expression on granular cells (including mast cells, eosinophils and neutrophils) and macrophages (MΦ) as indicated. The MCL+ fraction of granular cells likely reflects influx of neutrophils from the blood. MFI values are shown, calculated as the median of the fluorescence intensity. B, Resident peritoneal macrophages were cultured overnight with the indicated substances. Solid line histogram shows MCL staining. Grey filled histograms correspond to isotype controls.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3585393&req=5

pone-0057406-g003: Regulation of MCL surface expression under the effect of different stimuli.A, Rats were injected intraperitoneally with zymosan, peritoneal cells were obtained after 24 h and stained with mAbs for flow cytometry analysis. Discontinuous line histogram shows staining in the zymosan treated group, dotted line histogram shows MCL staining in the PBS control group. Histograms display MCL expression on granular cells (including mast cells, eosinophils and neutrophils) and macrophages (MΦ) as indicated. The MCL+ fraction of granular cells likely reflects influx of neutrophils from the blood. MFI values are shown, calculated as the median of the fluorescence intensity. B, Resident peritoneal macrophages were cultured overnight with the indicated substances. Solid line histogram shows MCL staining. Grey filled histograms correspond to isotype controls.
Mentions: Receptor expression under inflammatory conditions was examined by inducing sterile peritonitis with injections of zymosan in the peritoneal cavity of rats. 24 h after treatment, cells were collected by lavage of the peritoneal cavity and analyzed by flow cytometry. In the zymosan treated animals, MCL positive granulocytes appeared, corresponding to an expected influx of neutrophils. MCL expression was in addition increased on macrophages (Figure 3A). These were isolated from the peritoneum, maintained in vitro either in M-CSF, IFNγ plus LPS, or in IL-4. MCL was clearly increased when exposed to IFNγ and pro-inflammatory stimuli from G– bacteria, but it decreased following culture in the presence of anti-inflammatory IL-4 (Figure 3B). Taken together, these data suggest that MCL plays a role in inflammatory responses to microorganisms.

Bottom Line: We found that MCL co-immunoprecipitated with the activating adaptor protein FcεRIγ in these cells.Moreover, beads coated with anti-MCL antibody increased phagocytosis in the RMW cells.Together, these observations indicate that rat MCL is a receptor that activates phagocytosis in myeloid cells under inflammatory conditions.

View Article: PubMed Central - PubMed

Affiliation: Department of Anatomy, Institute of Basic Medical Sciences, University of Oslo, Oslo, Norway. a.l.pascual@medisin.uio.no

ABSTRACT
Macrophage C-type lectin (MCL) is a membrane surface receptor encoded by the Antigen Presenting Lectin-like gene Complex (APLEC). We generated a mouse monoclonal antibody for the study of this receptor in the rat. We demonstrate that rat MCL is expressed on blood monocytes and neutrophils, as well as on several tissue macrophage populations, including alveolar and peritoneal cavity macrophages. We also demonstrate MCL expression on a subset of resident spleen macrophages. Immunohistochemistry analysis of the spleen showed staining specifically in the marginal zone and red pulp. Exposure to pro-inflammatory mediators or to yeast cell wall extract (zymosan) increased surface MCL expression on peritoneal macrophages. We characterized a rat myeloid cell line, RMW, which expresses high levels of MCL. We found that MCL co-immunoprecipitated with the activating adaptor protein FcεRIγ in these cells. Moreover, beads coated with anti-MCL antibody increased phagocytosis in the RMW cells. Together, these observations indicate that rat MCL is a receptor that activates phagocytosis in myeloid cells under inflammatory conditions.

Show MeSH
Related in: MedlinePlus