Gene transfer and expression in human neutrophils. The phox homology domain of p47^phoxtranslocates to the plasma membrane but not to the membrane of mature phagosomes

Abstract

Abstract

Background

Neutrophils are non-dividing cells with poor survival after isolation. Consequently, exogenous gene expression in neutrophils is challenging. We report here the transfection of genes and expression of active proteins in human primary peripheral neutrophils using nucleofection.

Results

Exogenous gene expression in human neutrophils was achieved 2 h post-transfection. We show that neutrophils transfected by nucleofection are functional cells, able to respond to soluble and particulate stimuli. They conserved the ability to undergo physiological processes including phagocytosis. Using this technique, we were able to show that the phox homology (PX) domain of p47 ^phox localizes to the plasma membrane in human neutrophils. We also show that RhoB, but not the PX domain of p47 ^phox , is translocated to the membrane of mature phagosomes.

Conclusion

We demonstrated that cDNA transfer and expression of exogenous protein in human neutrophils is compatible with cell viability and is no longer a limitation for the study of protein function in human neutrophils.