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Monoclonal antibodies against the human mannose receptor that inhibit the binding of tissue-type plasminogen activator

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Author: Barrett-Bergshoeff, M.M. · Moorman, F. · Bos, R. · Rijken, D.C.
Type:article
Date:1997
Institution: Gaubius Instituut TNO
Source:Thrombosis and Haemostasis, 4, 77, 718-724
Identifier: 233874
doi: doi:10.1055/s-0038-1656040
Keywords: Biology · Antibodies, Monoclonal · Antigen-Antibody Reactions · Blotting, Western · Enzyme-Linked Immunosorbent Assay · Epitopes · Flow Cytometry · Humans · Immunohistochemistry · Lectins, C-Type · Macrophages · Mannose-Binding Lectins · Placenta · Receptors, Cell Surface · Tissue Plasminogen Activator

Abstract

To study the role of the mannose receptor in cellular uptake and degradation of tissue-type plasminogen activator (t-PA), a set of five monoclonal antibodies (Moab) was generated against the mannose receptor isolated from human placental tissue. All Moab specifically recognised the 175 kDa mannose receptor in a crude placenta extract, as shown in Western blot analysis. By use of immunohistochemistry, we showed that in human placenta only the Hofbauer cells (fetal macrophages) express the mannose receptor. Epitope competition experiments indicated that the Moab bound to at least two different epitopes on the receptor molecule. Moab 14-3, 14-5, and 15-2, which are directed against one of these epitopes, strongly inhibited the interaction between the purified mannose receptor and t-PA. These Moab also inhibited mannose receptor-mediated degradation of t-PA by cultured human macrophages. The low density lipoprotein receptor-related protein (LRP) mediated t-PA degradation was not affected by the Moab. It is concluded that the Moab are useful for studying the expression of the human mannose receptor in Western blot and in immunohistochemistry, and for studying the interactions between the human mannose receptor and the mannose-containing ligand t-PA. Copyright © 1997 Schattauer Verlag. Chemicals/CAS: Antibodies, Monoclonal; Epitopes; Lectins, C-Type; mannose receptor; Mannose-Binding Lectins; Receptors, Cell Surface; Tissue Plasminogen Activator, EC 3.4.21.68