Transduction of CD34+ cells with lentiviral vectors enables the production of large quantities of transgene-expressing immature and mature dendritic cells

✍ Scribed by Patrick Salmon; Jean-François Arrighi; Vincent Piguet; Bernard Chapuis; Rudolf H. Zubler; Didier Trono; Vincent Kindler

• Publisher: John Wiley and Sons
• Year: 2001
• Tongue: English
• Weight: 266 KB
• Volume: 3
• Category: Article
• ISSN: 1099-498X
• DOI: 10.1002/1521-2254(200107/08)3:4<311::aid-jgm198>3.0.co;2-b

✦ Synopsis

Background Genetically engineered dendritic cells (DC) presenting specific antigens to T cells may be of great interest for immunotherapy. For this reason, the production of transgene-expressing DC derived from CD34+ cells transduced either shortly after ex vivo purification or during their differentiation into DC were evaluated.

Methods CD34+ cells were transduced with lentivectors encoding for GFP before or after 21 days of culture with FLT3-ligand, thrombopoietin and stem cell factor and induction into DC with GM-CSF+IL-4 (G4) or G4+TNF (GT4). GFP and DC-specific marker expression was assessed by flow cytometry, and allostimulatory capacity was evaluated on GFP+ and GFPx sorted cells.

Results Immature (G4-induced) DC obtained from amplified CD34+ cells were transducible by lentiviral vectors while mature (GT4-induced) DC were rather refractory. Moreover, since differentiated DC did not proliferate, large quantities of vectors were required to generate transgene-expressing cells with this protocol. In contrast, greater numbers of both immature and mature GFPxexpressing DC were obtained with CD34+ cells exposed to lentivector shortly after purification. By the time of DC induction, GFP+ cells had increased by approximately 170-fold. After DC induction with G4, 32% of CD1a+, HLA-DR+, or CD40+ cells expressed GFP. CD1a+E-cadherin+ GFP+ Langerhans-like DC were also obtained. Incubation with TNF induced mature CD83+GFP+ DC that displayed a higher allostimulatory capacity than cells induced with G4 alone.

Conclusion

The transduction of a small number of CD34+ cells with minimal doses of lentivector may allow for the production of a large number of DC expressing selected antigens useful for immunotherapy.