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Blocking surface receptors on human monocyte derived macrophages

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ak4209

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Hello,

I've been culturing monocyte derived macrophages and exposing them to carbon particulates. I want to extend the study to look at which receptors are involved in the phagocytosis of particulates but not sure how to go about blocking receptors to see what effective that has on carbon loading of the cell.

I'm very new to this game and would therefore really appreciate any tips anybody has.

Thanks

Amy Knott

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 Posted Oct 26, 2007, 21:24 PM
Tracy

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Check these papers:
http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=1567749
Suppression of alveolar macrophage membrane receptor-mediated phagocytosis by model and actual particle-adsorbate complexes. Initial contact with the alveolar macrophage membrane.
G J Jakab, T H Risby, S S Sehnert, R R Hmieleski, and J E Farrington
Department of Environmental Health Sciences, Johns Hopkins University, School of Hygiene and Public Health, Baltimore, MD 21205.


http://www3.interscience.wiley.com/cgi-bin/abstract/112146442/ABSTRACT?CRETRY=1&SRETRY=0
Murine macrophage scavenger receptor: in vivo expression and function as receptor for macrophage adhesion in lymphoid and non-lymphoid organs
Derralynn A. Hughes *, Iain P. Fraser, Siamon Gordon
Sir William Dunn School of Pathology, Oxford


*Correspondence to Derralynn A. Hughes, Sir William Dunn School of Pathology, South Parks Rd, Oxford, OX1 3RE, GB (Fax: +44 865275501)


Keywords
Macrophage Scavenger receptor Adhesion


Abstract
Macrophage scavenger receptors are trimeric integral membrane glycoproteins which have been implicated in various macrophage functions including uptake of oxidized lipoprotein and the serum-dependent, divalent cation-independent adhesion of macrophages to tissue culture-treated plastic. In this study we have used a recently defined monoclonal antibody (2F8) which recognizes murine macrophage scavenger receptor, to explore its expression in lymphoid and non-lymphoid organs of the normal adult. Scavenger receptor was detected in the red pulp and marginal zone of normal adult mouse spleen, medulla of the thymus and subcapsular region of lymph nodes. Kupffer cells in the liver, alveolar macrophages in the lung and lamina propria macrophages in the gut all reacted with 2F8 monoclonal antibody. The antigen was not detected on any nonmacrophage cells, with the exception of sinusoidal endothelial cells in the liver. In the spleen, lymph node and liver, scavenger receptor antigen expression was associated specifically with phagocytic cells which had taken up colloidal carbon.
To examine macrophage adhesion in a context relevant to the interactions occurring within lymphoid and non-lymphoid organs, and the contribution of macrophage scavenger receptor to this adhesion, we designed an assay of macrophage adhesion to frozen tissue sections. Adhesion to most tissues was high and uniform in the absence of any chelating agents. The chelation of Ca2+ and Mg2+ revealed specific patterns of macrophage adhesion in lymphoid and non-lymphoid organs which was completely inhibited by 2F8. The ability of this antibody to block the EDTA-resistant adhesion correlated with tissue expression of the antigen in some tissues. Unlike adhesion to tissue culture-treated plastic, macrophage scavenger receptor-dependent adhesion of macrophages to frozen tissue sections did not exhibit an absolute requirement for exogenous fetal bovine serum indicating the presence of an endogenous ligand for scavenger receptor within the tissues. We propose that macrophage scavenger receptor is a candidate homing or retention molecule for macrophage localization within ligand-rich tissues.


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Posted Oct 28, 2007, 21:11 PM
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