Efficient and Robust NK-Cell Transduction With Baboon Envelope Pseudotyped Lentivector

• Published in: Frontiers in immunology Vol. 10; p. 2873
• Main Authors: Colamartino, Aurelien B L, Lemieux, William, Bifsha, Panojot, Nicoletti, Simon, Chakravarti, Nitin, Sanz, Joaquín, Roméro, Hugo, Selleri, Silvia, Béland, Kathie, Guiot, Mélanie, Tremblay-Laganière, Camille, Dicaire, Renée, Barreiro, Luis, Lee, Dean A, Verhoeyen, Els, Haddad, Elie
• Format: Journal Article
• Language: English
• Published: Switzerland Frontiers Media S.A 16.12.2019
• Subjects: baboon retrovirus envelope pseudotyped lentivectors; chimeric antigen receptor; cytotoxicity; Immunology; NK amplification and expansion system (NKAES); NK-cell transduction; NK amplification and expansion system (NKAES); baboon retrovirus envelope pseudotyped lentivectors; chimeric antigen receptor; NK-cell transduction; cytotoxicity
• ISSN: 1664-3224; 1664-3224
• DOI: 10.3389/fimmu.2019.02873

NK-cell resistance to transduction is a major technical hurdle for developing NK-cell immunotherapy. By using Baboon envelope pseudotyped lentiviral vectors (BaEV-LVs) encoding eGFP, we obtained a transduction rate of 23.0 ± 6.6% (mean ± SD) in freshly-isolated human NK-cells (FI-NK) and 83.4 ± 10.1% (mean ± SD) in NK-cells obtained from the NK-cell Activation and Expansion System (NKAES), with a sustained transgene expression for at least 21 days. BaEV-LVs outperformed Vesicular Stomatitis Virus type-G (VSV-G)-, RD114- and Measles Virus (MV)- pseudotyped LVs ( < 0.0001). mRNA expression of both BaEV receptors, ASCT1 and ASCT2, was detected in FI-NK and NKAES, with higher expression in NKAES. Transduction with BaEV-LVs encoding for CAR-CD22 resulted in robust CAR-expression on 38.3 ± 23.8% (mean ± SD) of NKAES cells, leading to specific killing of NK-resistant pre-B-ALL-RS4;11 cell line. Using a larger vector encoding a dual CD19/CD22-CAR, we were able to transduce and re-expand dual-CAR-expressing NKAES, even with lower viral titer. These dual-CAR-NK efficiently killed both CD19 - and CD22 -RS4;11 cells. Our results suggest that BaEV-LVs may efficiently enable NK-cell biological studies and translation of NK-cell-based immunotherapy to the clinic.