Geering Truffer, Barbara; Schmidt-Mende, Jan; Federzoni, Elena; Stoeckle, Christina; Simon, Hans-Uwe (2011). Protein overexpression following lentiviral infection of primary mature neutrophils is due to pseudotransduction. Journal of immunological methods, 373(1-2), pp. 209-18. Amsterdam: Elsevier 10.1016/j.jim.2011.08.024
Full text not available from this repository. (Request a copy)Neutrophils are terminally differentiated cells with a short life-span due to constitutive apoptosis. Because of these characteristics, genetic manipulation of neutrophils has been difficult, although it is highly desired given the importance of neutrophils in the immune system. Here we demonstrate that transduction of primary human mature neutrophils with enhanced green fluorescent protein (eGFP)-encoding lentiviral particles results in GFP-containing cells as previously reported. Yet, our data further show that GFP expression in neutrophils upon transduction is largely due to protein transfer, a process called lentiviral pseudotransduction, and not due to bona fide transduction. Thus, inhibition of viral genome integration by the reverse transcriptase inhibitor 3'-azido-3'-deoxythymidine (AZT) or of protein biosynthesis by cycloheximide (CHX) did not abolish GFP levels in transduced neutrophils. Importantly, lentiviral pseudotransduction of the enzyme death-associated protein kinase 2 (DAPK2) into primary human mature neutrophils resulted in increased protein levels, but not enzymatic functionality. Based on our data and previous reports of unspecific viral effects on immune cells following lentiviral transduction, we discourage scientists to use lentiviral transduction methods to manipulate primary mature neutrophils.
Item Type: |
Journal Article (Original Article) |
|---|---|
Division/Institute: |
04 Faculty of Medicine > Pre-clinic Human Medicine > Institute of Pharmacology |
UniBE Contributor: |
Geering Truffer, Barbara, Merz, Christina, Simon, Hans-Uwe |
Subjects: |
600 Technology > 610 Medicine & health |
ISSN: |
0022-1759 |
Publisher: |
Elsevier |
Language: |
English |
Submitter: |
Factscience Import |
Date Deposited: |
04 Oct 2013 14:18 |
Last Modified: |
05 Dec 2022 14:04 |
Publisher DOI: |
10.1016/j.jim.2011.08.024 |
PubMed ID: |
21925181 |
Web of Science ID: |
000297395400023 |
URI: |
https://boris.unibe.ch/id/eprint/5495 (FactScience: 210243) |
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