Comparative Analysis of Transduced Primary Human Dendritic Cells Generated by the Use of Three Different Lentiviral Vector Systems |
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Authors: | Elena Grabski Zoe Waibler Silke Schüle Björn-Philipp Kloke Linda Y Sender Sylvia Panitz Klaus Cichutek Matthias Schweizer Ulrich Kalinke |
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Institution: | (1) Institute for Experimental Infection Research, TWINCORE, Centre for Experimental and Clinical Infection Research, Feodor-Lynen-Str. 7, 30625 Hannover, Germany;(2) Junior Research Group NG2, Paul-Ehrlich-Institut, 63225 Langen, Germany;(3) Division of Medical Biotechnology, Paul-Ehrlich-Institut, 63225 Langen, Germany;(4) Division of Immunology, Paul-Ehrlich-Institut, 63225 Langen, Germany; |
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Abstract: | Lentiviral gene transfer vectors are suitable for genetically modifying non-cycling primary human cells. In this study, we
analyzed transduced human dendritic cells (DC) generated by the use of three different GFP-encoding lentiviral vectors, HIV-2
ROD A Δenv-GFP (ROD A), SIVsmm PBj ΔE EGFP (PBj), and SIVmac ΔE EGFP (SIVmac). CD14+ monocytes were isolated from buffy coat, transduced, and differentiated to immature and mature DC. Cytofluometric analysis
of DC revealed high transduction efficiencies at MOI 1 for simian immunodeficiency virus (SIV)-derived vectors PBj and SIVmac
ranging between 80–90 and 70–90%, respectively. In contrast, transduction with ROD A resulted only in approximately 30%-positive
DC at the same MOI. Of note, none of the analyzed vectors affected expression of maturation and/or activation markers. Moreover,
transduction with PBj or SIVmac did not induce significant cytokine responses whereas ROD A transduction stimulated weak interferon-alpha
responses. SIVmac transduced DC showed normal phagocytosis of antigen and normal allo T cell stimulatory capacity when compared
with untreated DC. Thus, the SIVmac lentiviral transduction vector is suitable for efficient genetic modification of human
DC without affecting phenotype or function and thus qualifies this vector as a versatile tool for use in basic research. |
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