Journal
JOURNAL OF IMMUNOLOGICAL METHODS
Volume 327, Issue 1-2, Pages 63-74Publisher
ELSEVIER
DOI: 10.1016/j.jim.2007.07.011
Keywords
eosinophils; detection; quantitation; siglec-F; flow cytometty
Categories
Funding
- NHLBI NIH HHS [R01 HL033391-22, R01 HL033391, R01 HL033391-27, R01 HL033391-21, R01 HL033391-18, R01 HL033391-24, R01 HL033391-23, R01 HL033391-19, R01 HL033391-20] Funding Source: Medline
- NIAID NIH HHS [R01 AI037293-13, R01 AI037293-09, R01 AI015608, R01 AI015608-25, R01 AI015608-28, T32 AI007046, R01 AI037293-11, R01 AI015608-27, R01 AI037293-10, R01 AI037293-14, R01 AI015608-29, R01 AI037293, R01 AI015608-26, R01 AI037293-12] Funding Source: Medline
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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 CD11c(low/-) staining profile. In the BAL fluid, this flow cytometry-based method of eosmophil identification/cluantitation 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. (c) 2007 Elsevier B.V. All rights reserved.
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