TY - JOUR
T1 - In vivo functional analysis and genetic modification of in vitro-derived mouse neutrophils
AU - McDonald, Jacqueline U
AU - Cortini, Andrea
AU - Rosas, Marcela
AU - Fossati-Jimack, Liliane
AU - Ling, Guang Sheng
AU - Lewis, Kimberley J
AU - Dewitt, Sharon
AU - Liddiard, Kate
AU - Brown, Gordon D
AU - Jones, Simon A
AU - Hallett, Maurice B
AU - Botto, Marina
AU - Taylor, Philip R
PY - 2011
Y1 - 2011
N2 - 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.
AB - 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.
KW - Inflammation
KW - Animal models
KW - Conditional imortalization
U2 - 10.1096/fj.10-178517
DO - 10.1096/fj.10-178517
M3 - Article
C2 - 21368104
SN - 1530-6860
VL - 25
SP - 2
EP - 11
JO - The FASEB Journal
JF - The FASEB Journal
ER -