HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in ISI Web of Science Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via ISI Web of Science (3) Citing Articles via Google Scholar Google Scholar Articles by Bunn, H.J. Articles by Grigg, J. Search for Related Content PubMed PubMed Citation Articles by Bunn, H.J. Articles by Grigg, J.

Applicability of laser scanning cytometry to study paediatric alveolar macrophages

¹ Dept of Child Health and ² Division of Respiratory Medicine, Glenfield Hospital, Institute for Lung Health, University of Leicester, UK

CORRESPONDENCE: J. Grigg, Senior Lecturer in Paediatric Respiratory Medicine, Leicester Children's Asthma Centre, University of Leicester, Leicester Royal Infirmary, LE2 7LX. Fax: 44 1162523282. E-mail: jg33@le.ac.uk

Keywords: alveolar macrophage, bronchoalveolar lavage, child, laser scanning cytometry

Received: April 25, 2002
Accepted July 16, 2002

J. Grigg was supported by Medisearch and Henry Smith's Charity.

Laser scanning cytometry (LSC) generates quantitative information on immune receptor expression from cells cytocentrifuged onto a microscope slide. In children, the description of developmental changes in immune receptor expression on alveolar macrophages (AM) has been limited by the small number of cells recovered by bronchoalveolar lavage (BAL). The applicability of LSC to the study of AM from normal children was therefore assessed.

AM were obtained by BAL of normal children following intubation prior to elective surgery. The ability of LSC to identify the cytoplasm of AM was assessed using either: 1) autofluorescence; 2) forward scatter; 3) nuclear staining with propidium iodide; or 4) a fluorescent-labelled monoclonal antibody to CD68, a pan-macrophage antigen.

LSC could only reliably identify individual AM when stained with CD68. The sensitivity for detecting single whole AM using CD68 was 0.97 and the positive predictive value was 0.88, respectively, with excellent repeatability. In addition, a range of immunofluorescence parameters were generated for CD68.

It is concluded that laser scanning cytometry is suited to the study of immune receptor expression from small numbers of paediatric alveolar macrophages, when CD68 is used for cell identification.