Enhanced lentiviral transduction of monocyte-derived dendritic cells in the presence of conditioned medium from dying monocytes.
Lentiviral vectors (LVs) are attractive vehicles for the transduction of human dendritic cells (DCs) in order to mobilize their endogenous antigen presentation pathways. We analyzed here how to improve the efficiency of LV transduction, which we performed at the initial stages of the differentiation of purified monocytes into dendritic cells (Mo-DCs). Using LVs pseudotyped with the vesicular stomatitis virus envelope G glycoprotein (VSV-G), we found that a conditioned medium derived from dying monocytes (MCM) improved by 2- to 10- fold the proportion of transduced Mo-DCs. This enhanced transduction efficiency requires the presence of MCM during the initial stage of LV transduction and does not affect the phenotype and antigen presentation function of terminally differentiated Mo-DCs. Importantly, we found that MCM derived from a human acute monocytic leukemia cell line, THP-1, was equally effective. The MCM activity was heat stable (56 degrees C) and was present in the soluble fraction after high-speed centrifugation. Altogether our results show that a soluble factor present in dying monocyte cultures can replace advantageously facilitating agents such as Polybrene, to achieve high LV transductions levels. This protocol can be performed with autologous monocytes and is therefore applicable in clinical settings.
Masurier
C
C
Laboratoire d'Immunologie, Généthon, CNRS UMR 8115, 91002 Evry Cedex, France. masurier@genethon.fr
Boutin
S
S
Veron
P
P
Bernard
J
J
Danos
O
O
Davoust
J
J
D016923:Cell Death; D002478:Cells, Cultured; D017077:Culture Media, Conditioned / Q000494:pharmacology*; D003713:Dendritic Cells / Q000166:cytology* / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D016086:Lentivirus / Q000235:genetics*; D009000:Monocytes / Q000166:cytology* / Q000378:metabolism*; D014161:Transduction, Genetic / Q000379:methods*
D017077:Culture Media, Conditioned