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

McDonald, Jacqueline U., Cortini, Andrea, Rosas, Marcela, Fossati-Jimack, Liliane, Ling, Guang Sheng, Lewis, Kimberley, Dewitt, Sharon, Liddiard, Kate, Brown, Gordon D., Jones, Simon Arnett, Hallett, Maurice Bartlett, Botto, Marina and Taylor, Philip Russel 2011. In vivo functional analysis and genetic modification of in vitro-derived mouse neutrophils. The FASEB Journal 25 (6) , pp. 1972-1982. 10.1096/fj.10-178517

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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.

Item Type: Article
Date Type: Publication
Status: Published
Schools: Medicine
Systems Immunity Research Institute (SIURI)
Subjects: Q Science > QH Natural history > QH426 Genetics
Q Science > QR Microbiology > QR180 Immunology
Uncontrolled Keywords: inflammation; animal models; conditional immortalization
Publisher: Federation of American Society of Experimental Biology
ISSN: 0892-6638
Last Modified: 08 Oct 2018 21:21
URI: http://orca.cf.ac.uk/id/eprint/26419

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