Detection and quantitation of eosinophils in the murine respiratory tract by flow cytometry

Whitney W. Stevens, Taeg S. Kim, Lindsey M. Pujanauski, Xueli Hao, Thomas J. Braciale*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

119 Scopus citations

Abstract

Traditionally, the identification and quantification of eosinophils in inflammatory tissues and exudates has been primarily based upon morphologic criteria and manual counting. In this study, we describe a new flow cytometry-based assay to enumerate eosinophils present in murine bronchoalveolar lavage fluid (BAL) and lung parenchyma obtained from the normal/non-inflamed respiratory tract, following experimentally-induced allergic pulmonary inflammation, and during experimental infection with respiratory syncytial virus (RSV). By using a murine Siglec-F-specific antibody in combination with antibodies directed to CD45 and CD11c, we demonstrate that eosinophils can be distinguished from other cell types in the BAL fluid and lung parenchyma based upon their distinct CD45+ Siglec-F+ and CD11clow/- staining profile. In the BAL fluid, this flow cytometry-based method of eosinophil identification/quantitation yields results comparable to the standard morphology-based method without the potential observer bias or staining artifacts inherent in morphology-based quantitation. Furthermore, this flow cytometry-based method can be directly adapted to enumerate eosinophils infiltrating the inflamed lung parenchyma, thereby obviating the need for quantitative morphometry of tissue sections.

Original languageEnglish (US)
Pages (from-to)63-74
Number of pages12
JournalJournal of Immunological Methods
Volume327
Issue number1-2
DOIs
StatePublished - Oct 31 2007

Keywords

  • Detection
  • Eosinophils
  • Flow cytometry
  • Quantitation
  • Siglec-F

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

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