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

McDonald, Jacqueline U., Cortini, Andrea, Rosas, Marcela ORCID: https://orcid.org/0000-0002-9442-9638, Fossati-Jimack, Liliane, Ling, Guang Sheng, Lewis, Kimberley, Dewitt, Sharon ORCID: https://orcid.org/0000-0001-8169-8241, Liddiard, Kate ORCID: https://orcid.org/0000-0002-0953-1997, Brown, Gordon D., Jones, Simon Arnett ORCID: https://orcid.org/0000-0001-7297-9711, Hallett, Maurice Bartlett ORCID: https://orcid.org/0000-0001-8197-834X, Botto, Marina and Taylor, Philip Russel ORCID: https://orcid.org/0000-0003-0163-1421 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:	06 Mar 2024 07:27
URI:	https://orca.cardiff.ac.uk/id/eprint/26419

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