In vivo functional analysis and genetic modification of in vitro-derived mouse neutrophils

Jacqueline U McDonald, Andrea Cortini, Marcela Rosas, Liliane Fossati-Jimack, Guang Sheng Ling, Kimberley J Lewis, Sharon Dewitt, Kate Liddiard, Gordon D Brown, Simon A Jones, Maurice B Hallett, Marina Botto, Philip R Taylor

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

Mature neutrophils are notoriously short-lived 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.-McDonald, J. U., Cortini, A., Rosas, M., Fossati-Jimack, L., Ling, G. S., Lewis, K. J., Dewitt, S., Liddiard, K., Brown, G. D., Jones, S. A., Hallett, M. B., Botto, M., Taylor, P. R. In vivo functional analysis and genetic modification of in vitro-derived mouse neutrophils.
Original languageEnglish
Pages (from-to)2-11
Number of pages11
JournalThe FASEB Journal
Volume25
DOIs
Publication statusPublished - 2011

Keywords

  • Inflammation
  • Animal models
  • Conditional imortalization

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