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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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Immunostaining of rat spleen.Serial-cut frozen sections were stained with mAbs towards A, rat MCL B, rat CD169 C, rat MHC class II D, human MHC class I (negative control) and visualized with peroxidase-conjugated secondary antibody and DAB substrate. RP: red pulp. PALS: periarteriolar lymphoid sheath. FOLL: follicle. MZ: marginal zone.
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pone-0057406-g002: Immunostaining of rat spleen.Serial-cut frozen sections were stained with mAbs towards A, rat MCL B, rat CD169 C, rat MHC class II D, human MHC class I (negative control) and visualized with peroxidase-conjugated secondary antibody and DAB substrate. RP: red pulp. PALS: periarteriolar lymphoid sheath. FOLL: follicle. MZ: marginal zone.

Mentions: Because the WEN42 mAb only stained a fraction of macrophages in splenic cell suspensions, we investigated the localization of MCL+ and MCL– cells in the spleen by immunohistochemistry. With WEN42, there was little or no staining of the white pulp (follicles and periarteriolar lymphoid sheaths, PALS), but we could observe distinct staining of cells in the marginal zone (MZ), probably MZ macrophages, including the thin marginal sinus layer of CD169+ MZ metallophilic macrophages at the border between the MZ and the white pulp. There was also abundant, although weaker, staining of the red pulp, probably red pulp macrophages (Figure 2).


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)

Immunostaining of rat spleen.Serial-cut frozen sections were stained with mAbs towards A, rat MCL B, rat CD169 C, rat MHC class II D, human MHC class I (negative control) and visualized with peroxidase-conjugated secondary antibody and DAB substrate. RP: red pulp. PALS: periarteriolar lymphoid sheath. FOLL: follicle. MZ: marginal zone.
© Copyright Policy
Related In: Results  -  Collection

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

pone-0057406-g002: Immunostaining of rat spleen.Serial-cut frozen sections were stained with mAbs towards A, rat MCL B, rat CD169 C, rat MHC class II D, human MHC class I (negative control) and visualized with peroxidase-conjugated secondary antibody and DAB substrate. RP: red pulp. PALS: periarteriolar lymphoid sheath. FOLL: follicle. MZ: marginal zone.
Mentions: Because the WEN42 mAb only stained a fraction of macrophages in splenic cell suspensions, we investigated the localization of MCL+ and MCL– cells in the spleen by immunohistochemistry. With WEN42, there was little or no staining of the white pulp (follicles and periarteriolar lymphoid sheaths, PALS), but we could observe distinct staining of cells in the marginal zone (MZ), probably MZ macrophages, including the thin marginal sinus layer of CD169+ MZ metallophilic macrophages at the border between the MZ and the white pulp. There was also abundant, although weaker, staining of the red pulp, probably red pulp macrophages (Figure 2).

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