SpringerProtocols: Abstract: Generation of Retroviral Macrophage cDNA Expression Libraries and Functional Screening for Surface Receptors

1. Generation of Retroviral Macrophage cDNA Expression Libraries and Functional Screening for Surface Receptors

By: Georgia Schäfer², Gordon D. Brown*²

Abstract

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Macrophages, tissue-based phagocytic cells derived from blood monocytes, play important roles in immunity and homeostasis. As professional scavengers, macrophages phagocytose microbes, apoptotic and necrotic cells and take up modified lipoprotein particles. However, many of their complex interactions with other immune cells and/or various ligands are not yet clearly understood. To identify and isolate macrophage cell surface molecules, particularly macrophage receptors, for which ligands are known, a powerful generalized screening method has been established. As discussed in this chapter, this technique based on function has been successfully applied for the identification of dectin-1, the major macrophage receptor involved in the binding and recognition of β-glucans (Nature 413:36–37, 2001).

Affiliation(s): (2) Institute for Infectious Diseases and Molecular Medicine, Faculty of Health Sciences, University of Cape Town, Observatory 7925, Cape Town, South Africa

Book Title: Macrophages and Dendritic Cells: Methods and Protocols

Series: Methods in Molecular Biology | Volume: 531 | Pub. Date: Jul-01-2008 | Page Range: 1-15 | DOI: 10.1007/978-1-59745-396-7_1

Subject: Immunology

Key Words: cDNA expression cloning - Retroviral cDNA expression library - Functional screening - Surface molecule - Ligand binding - Receptor cloning - Stable transfection - Fluorescent microscopy

References

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