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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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RMW is a rat myeloid cell line. A–DTEM ultrastructure of RMW cells. M: mitochondria. LB: lipid bodies. RER: rough endoplasmic reticulum. MV: multivesicular endosomes. G, Golgi. E, Flow cytometry analysis of RMW surface markers. Solid lines show the indicated surface markers; grey filled histograms correspond to isotype control.
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pone-0057406-g004: RMW is a rat myeloid cell line. A–DTEM ultrastructure of RMW cells. M: mitochondria. LB: lipid bodies. RER: rough endoplasmic reticulum. MV: multivesicular endosomes. G, Golgi. E, Flow cytometry analysis of RMW surface markers. Solid lines show the indicated surface markers; grey filled histograms correspond to isotype control.

Mentions: The RMW cell line was derived by in vitro culture of PVG rat splenocytes as described [17]. Whereas the rat macrophage cell line R2 [22] did not express MCL (data not shown), RMW cells were MCL+ by flow cytometry. This cell line was considered a useful tool for further studies and was therefore characterized more thoroughly. Transmission electron microscopy analysis of the ultrastructure of RMW cells (Figure 4 A–D) showed a ruffled plasma membrane, suggesting a role for sensing and probing the extracellular environment. The nucleus was eccentrically located and indented, with a prominent nucleolus in many cells. The cytoplasm contained several mitochondria, and in most cells varying modest amounts of rough endoplasmic reticulum. Golgi complexes were distinct but not particularly prominent. Cytoplasmic vesicular structures were observed, including multivesicular bodies, and in some cells large structures with morphological characteristics of lipid bodies.


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)

RMW is a rat myeloid cell line. A–DTEM ultrastructure of RMW cells. M: mitochondria. LB: lipid bodies. RER: rough endoplasmic reticulum. MV: multivesicular endosomes. G, Golgi. E, Flow cytometry analysis of RMW surface markers. Solid lines show the indicated surface markers; grey filled histograms correspond to isotype control.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC3585393&req=5

pone-0057406-g004: RMW is a rat myeloid cell line. A–DTEM ultrastructure of RMW cells. M: mitochondria. LB: lipid bodies. RER: rough endoplasmic reticulum. MV: multivesicular endosomes. G, Golgi. E, Flow cytometry analysis of RMW surface markers. Solid lines show the indicated surface markers; grey filled histograms correspond to isotype control.
Mentions: The RMW cell line was derived by in vitro culture of PVG rat splenocytes as described [17]. Whereas the rat macrophage cell line R2 [22] did not express MCL (data not shown), RMW cells were MCL+ by flow cytometry. This cell line was considered a useful tool for further studies and was therefore characterized more thoroughly. Transmission electron microscopy analysis of the ultrastructure of RMW cells (Figure 4 A–D) showed a ruffled plasma membrane, suggesting a role for sensing and probing the extracellular environment. The nucleus was eccentrically located and indented, with a prominent nucleolus in many cells. The cytoplasm contained several mitochondria, and in most cells varying modest amounts of rough endoplasmic reticulum. Golgi complexes were distinct but not particularly prominent. Cytoplasmic vesicular structures were observed, including multivesicular bodies, and in some cells large structures with morphological characteristics of lipid bodies.

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