水泡性口炎病毒
狒狒
生物
转导(生物物理学)
CD19
病毒学
麻疹病毒
免疫疗法
细胞培养
分子生物学
抗原
免疫学
病毒
免疫系统
遗传学
生物化学
内分泌学
麻疹
接种疫苗
作者
Colamartino Ab,William Lemieux,Panojot Bifsha,Simon Nicoletti,Nitin Chakravarti,Remon Js,H Roméro,Silvia Selleri,Kathie Béland,Mélanie Guiot,Camille Tremblay‐Laganière,Renée Dicaire,Luis B. Barreiro,Lee Da,Els Verhoeyen,Elias K. Haddad
摘要
ABSTRACT 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% in freshly-isolated NK-cells (FI-NK) and 83.4±10.1% in NK-cells obtained from the NK-cell Activation and Expansion System (NKAES), even at low MOI, 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 (p<0.001). mRNA expression of both BaEV receptors, ASCT1 and ASCT2, was detected in FI-NK and NKAES, with much higher expression in NKAES. Transduction with BaEV-LVs encoding for CAR-CD22 resulted in robust CAR-expression on 44.2%±14.2% of NKAES cells, which allowed the specific killing of the NK-resistant pre-B-ALL-RS4;11 cell line. Using a larger vector, encoding a dual CD19/CD22-CAR separated by T2A, we were able to transduce and re-expand dual-CAR-expressing NKAES, even with low viral titer. These dual-CAR-NK efficiently and specifically killed both CD19 KO -and CD22 KO -RS4;11 cells, which may overcome antigen-loss escape in the clinical setting. Our results suggest that BaEV-LVs may efficiently enable NK-cell biological studies and translation of NK-cell-based immunotherapy to the clinic.
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