A clinically adaptable method to enhance the cytotoxicity of natural killer cells against B-cell malignancies

Abstract Background aims Retroviral transduction of anti-CD19 chimeric antigen receptors significantly enhances the cytotoxicity of natural killer (NK) cells against B-cell malignancies. We aimed to validate a more practical, affordable and safe method for this purpose. Methods We tested the express...

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Veröffentlicht in:Cytotherapy (Oxford, England) England), 2012-08, Vol.14 (7), p.830-840
Hauptverfasser: Shimasaki, Noriko, Fujisaki, Hiroyuki, Cho, Duck, Masselli, Marika, Lockey, Timothy, Eldridge, Paul, Leung, Wing, Campana, Dario
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Sprache:eng
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Zusammenfassung:Abstract Background aims Retroviral transduction of anti-CD19 chimeric antigen receptors significantly enhances the cytotoxicity of natural killer (NK) cells against B-cell malignancies. We aimed to validate a more practical, affordable and safe method for this purpose. Methods We tested the expression of a receptor containing CD3ζ and 4-1BB signaling molecules (anti-CD19-BB-ζ) in human NK cells after electroporation with the corresponding mRNA using a clinical-grade electroporator. The cytotoxic capacity of the transfected NK cells was tested in vitro and in a mouse model of leukemia. Results Median anti-CD19-BB-ζ expression 24 h after electroporation was 40.3% in freshly purified ( n =18) and 61.3% in expanded ( n = 31) NK cells; median cell viability was 90%. NK cells expressing anti-CD19-BB-ζ secreted interferon (IFN)-γ in response to CD19-positive target cells and had increased cytotoxicity. Receptor expression was detectable 6 h after electroporation, reaching maximum levels at 24–48 h; specific anti-CD19 cytotoxicity was observed at 96 h. Levels of expression and cytotoxicities were comparable with those achieved by retroviral transduction. A large-scale protocol was developed and applied to expanded NK cells (median NK cell number 2.5 × 108 , n = 12). Median receptor expression after 24 h was 82.0%; NK cells transfected under these conditions exerted considerable cytotoxicity in xenograft models of B-cell leukemia. Conclusions The method described here represents a practical way to augment the cytotoxicity of NK cells against B-cell malignancies. It has the potential to be extended to other targets beyond CD19 and should facilitate the clinical use of redirected NK cells for cancer therapy.
ISSN:1465-3249
1477-2566
DOI:10.3109/14653249.2012.671519