Accuri C6 Flow Cytometer System
Originally published online as doi:10.1189/jlb.1108699 on April 29, 2009

Published online before print April 29, 2009
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(Journal of Leukocyte Biology. 2009;86:479-489.)
© 2009 Society for Leukocyte Biology

Pivotal Advance: Expansion of small sputum macrophages in CF: failure to express MARCO and mannose receptors

Adam K. A. Wright*, Satish Rao*,{dagger}, Simon Range{ddagger}, Christiane Eder§, Thomas P. J. Hofer§, Marion Frankenberger§, Lester Kobzik||, Chris Brightling*,{ddagger}, Jonathan Grigg*,¶ and Loems Ziegler-Heitbrock*,§,1

* Department of Infection, Immunity and Inflammation, University of Leicester, Leicester, United Kingdom;
{dagger} Department of Respiratory Medicine and Cystic Fibrosis, Birmingham Children’s Hospital, Birmingham, United Kingdom;
{ddagger} Institute for Lung Health, Glenfield Hospital, University Hospitals of Leicester National Health Service, Leicester, United Kingdom;
§ Clinical Cooperation Group, Inflammatory Lung Diseases, Asklepios-Fachklinik and Helmholtz Zentrum München, German Research Center for Environmental Health, Gauting, Germany;
|| Physiology Program, Harvard School of Public Health, Boston, Massachusetts, USA; and
Academic Unit of Paediatrics, Institute of Cell and Molecular Science, Queen Mary University London, London, United Kingdom

1. Correspondence: Helmholtz Zentrum München, German Research Center for Environmental Health, Robert-Koch-Allee 29, 82131 Gauting, Germany. E-mail: zeigler-heitbrock{at}helmholtz-muenchen.de

ABSTRACT

Macrophages in the airways form an important element of immune defense and inflammation. We analyzed induced sputum from airways of patients with CF for the types of macrophages present, their receptor expression, and phagocytic function. In samples from patients and age-matched controls, macrophages were analyzed by multicolor flow cytometry, scavenger receptor expression was studied at the protein and mRNA level, and receptor function was investigated using fluorescent particles. In adult patients with CF, we discovered a pronounced expansion of the small CD14+ DR+ CD68weak+ macrophages to 73 ± 18% compared with 16 ± 8% in healthy controls. Expression of the MARCO and CD206 (mannose receptor) was strongly reduced at the mRNA and protein level in sputum macrophages. Antibody-blocking studies showed that MARCO mediates phagocytosis of unopsonized particles. In line with reduced MARCO expression, sputum macrophages in CF showed a deficient uptake of particles (23±9% of cells) compared with healthy controls (71±15%). The deficiency of MARCO expression in the predominant small sputum macrophages in CF may lead to impaired clearance of inhaled particles with increased inflammation and damage to the CF lung.

Key Words: cystic fibrosis • phagocytosis • scavenger receptor


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