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Publication Details
AFRICAN RESEARCH NEXUS
SHINING A SPOTLIGHT ON AFRICAN RESEARCH
In vivo functional analysis and genetic modification of in vitro-derived mouse neutrophils
FASEB Journal, Volume 25, No. 6, Year 2011
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Description
Mature neutrophils are notoriously shortlived immune cells that cannot be genetically manipulated. Analysis of gene function therefore requires genetically modified animals, which is expensive, time-consuming, and costly in animal life. Analysis of gene function in neutrophils in a physiologically relevant context thus represents a significant problem in the field. We sought to overcome this obstruction in the field by developing a strategy for the analysis of gene function in neutrophils in a physiologically relevant context. Here, we demonstrate the functional relevance of in vitro conditional-Hoxb8 immortalized precursor-derived neutrophils. In vitro-derived neutrophils functionally resembled primary neutrophils, but critically, neutrophils generated in this way can be adoptively transferred into live animals and tracked during inflammatory responses using singlecell analysis to define functional attributes. We have validated this approach using CD11b-deficient neutrophils and replicated the key findings observed in genetargeted animals and in naturally CD11b-deficient humans. Furthermore, we show that by retroviral transduction, one can generate stable alterations in the precursor cell lines and thus a continuous supply of functionally altered neutrophils. This novel technological advance offers for the first time the possibility of applying higher-throughput genetic modification and in vivo functional analysis to the neutrophil-lineage. © FASEB.
Authors & Co-Authors
McDonald, Jacqueline U.
United Kingdom, Cardiff
Cardiff University School of Medicine
Rosas, Marcela
United Kingdom, Cardiff
Cardiff University School of Medicine
Liddiard, Kate
United Kingdom, Cardiff
Cardiff University School of Medicine
Brown, Gordon D.A.
United Kingdom, Aberdeen
University of Aberdeen
Jones, Simon A.
United Kingdom, Cardiff
Cardiff University School of Medicine
Botto, Marina B.
United Kingdom, London
Imperial College London
Taylor, Philip Russel
United Kingdom, Cardiff
Cardiff University School of Medicine
Statistics
Citations: 34
Authors: 7
Affiliations: 3
Identifiers
Doi:
10.1096/fj.10-178517
ISSN:
15306860
Research Areas
Genetics And Genomics