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- Title
In vivo functional analysis and genetic modification of in vitro-derived mouse neutrophils.
- Authors
McDonald, Jacqueline U.; Cortini, Andrea; Rosas, Marcela; Fossati-Jimack, Liliane; Ling, Guang Sheng; Lewis, Kimberley J.; Dewitt, Sharon; Liddiard, Kate; Brown, Gordon D.; Jones, Simon A.; Hallett, Maurice B.; Botto, Marina; Taylor, Philip R.
- Abstract
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 single-cell analysis to define functional attributes. We have validated this approach using CD11b-deficient neutrophils and replicated the key findings observed in gene-targeted 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.
- Subjects
NEUTROPHILS; INFLAMMATION; TRANSGENIC organisms; CONDITIONAL immortality; ANIMAL models in research
- Publication
FASEB Journal, 2011, Vol 25, Issue 6, p1972
- ISSN
0892-6638
- Publication type
Article
- DOI
10.1096/fj.10-178517