Gene transfer and expression in human neutrophils. The phox homology domain of p47phox translocates to the plasma membrane but not to the membrane of mature phagosomes
| Autor: | Sergio D. Catz, Beverly A. Ellis, Agnieszka A. Brzezinska, Daniela B. Munafo, Jennifer L. Johnson |
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| Jazyk: | angličtina |
| Rok vydání: | 2006 |
| Předmět: |
lcsh:Immunologic diseases. Allergy
Neutrophils RHOB Immunology Nucleofection HL-60 Cells Biology Phagosomes Gene expression Humans Cells Cultured Phagosome NADPH oxidase Methodology Article Cell Membrane Gene Transfer Techniques NADPH Oxidases Cell Differentiation Transfection PX domain Molecular biology Transport protein Cell biology Protein Structure Tertiary Protein Transport biology.protein lcsh:RC581-607 |
| Zdroj: | BMC Immunology BMC Immunology, Vol 7, Iss 1, p 28 (2006) |
| ISSN: | 1471-2172 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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