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Surface-bound myeloperoxidase is a ligand for
recognition of late apoptotic neutrophils by human lung surfactant
proteins A and D |
| Anne Jäkel1,Robert B. Sim1,Howard Clark2,Kenneth B. M. Reid3, |
| 1.Department of Pharmacology,
University of Oxford, Mansfield Road, Oxford OX1 3QT, UK,；MRC Immunochemistry Unit,
Department of Biochemistry, University of Oxford, South Parks Road,
Oxford OX1 3QU, UK; 2.University of Southampton,
School of Medicine, Division of Infection Inflammation & Immunity,
Southampton General Hospital, Southampton SO16 6YD, UK；MRC Immunochemistry Unit,
Department of Biochemistry, University of Oxford, South Parks Road,
Oxford OX1 3QU, UK; 3.MRC Immunochemistry Unit,
Department of Biochemistry, University of Oxford, South Parks Road,
Oxford OX1 3QU, UK; |
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Abstract Surfactant proteins A (SP-A) and D (SP-D), both members of the collectin family, play a well established role in apoptotic cell recognition and clearance. Recent in vitro data show that SP-A and SP-D interact with apoptotic neutrophils in a distinct manner. SP-A and SP-D bind in a Ca2+-dependent manner to viable and early apoptotic neutrophils whereas the much greater interaction with late apoptotic neutrophils is Ca2+-independent. Cell surface molecules on the apoptotic target cells responsible for these interactions had not been identified and this study was done to find candidate target molecules. Myeloperoxidase (MPO), a specific intracellular defense molecule of neutrophils that becomes exposed on the outside of the cell upon apoptosis, was identified by affinity purification, mass-spectrometry and western blotting as a novel binding molecule for SP-A and SP-D. To confirm its role in recognition, it was shown that purified immobilised MPO binds SP-A and SP-D, and that MPO is surface-exposed on late apoptotic neutrophils. SP-A and SP-D inhibit binding of an anti-MPO monoclonal Ab to late apoptotic cells. Fluorescence microscopy confirmed that anti-MPO mAb and SP-A/SP-D colocalise on late apoptotic neutrophils. Desmoplakin was identified as a further potential ligand for SP-A, and neutrophil defensin as a target for both proteins.
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| Keywords
SP-A
SP-D
myeloperoxidase
neutrophils
flow cytometry
mass spectrometry
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Issue Date: 01 June 2010
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