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Journal of Immunology Research
Volume 2016, Article ID 9540975, 13 pages
http://dx.doi.org/10.1155/2016/9540975
Research Article

Luciferase mRNA Transfection of Antigen Presenting Cells Permits Sensitive Nonradioactive Measurement of Cellular and Humoral Cytotoxicity

1Division of Translational and Experimental Oncology, Department of Medicine III, Johannes Gutenberg University, Freiligrathstrasse 12, 55131 Mainz, Germany
2BioNTech Cell & Gene Therapies GmbH, An der Goldgrube 12, 55131 Mainz, Germany
3BioNTech AG, An der Goldgrube 12, 55131 Mainz, Germany
4Ganymed Pharmaceuticals AG, An der Goldgrube 12, 55131 Mainz, Germany

Received 15 September 2015; Accepted 10 December 2015

Academic Editor: Andrew Geall

Copyright © 2016 Tana A. Omokoko et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Immunotherapy is rapidly evolving as an effective treatment option for many cancers. With the emerging fields of cancer vaccines and adoptive cell transfer therapies, there is an increasing demand for high-throughput in vitro cytotoxicity assays that efficiently analyze immune effector functions. The gold standard 51Cr-release assay is very accurate but has the major disadvantage of being radioactive. We reveal the development of a versatile and nonradioactive firefly luciferase in vitro transcribed (IVT) RNA-based assay. Demonstrating high efficiency, consistency, and excellent target cell viability, our optimized luciferase IVT RNA is used to transfect dividing and nondividing primary antigen presenting cells. Together with the long-lasting expression and minimal background, the direct measurement of intracellular luciferase activity of living cells allows for the monitoring of killing kinetics and displays paramount sensitivity. The ability to cotransfect the IVT RNA of the luciferase reporter and the antigen of interest into the antigen presenting cells and its simple read-out procedure render the assay high-throughput in nature. Results generated were comparable to the 51Cr release and further confirmed the assay’s ability to measure antibody-dependent cell-mediated cytotoxicity and complement-dependent cytotoxicity. The assay’s combined simplicity, practicality, and efficiency tailor it for the analysis of antigen-specific cellular and humoral effector functions during the development of novel immunotherapies.