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Quantification and functional evaluation of CD40L production from the adenovirus vector ONCOS-401

Cancer Gene Therapy volume 26, pages 26–31 (2019)Cite this article

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Abstract

Adaptive immunity involves activation of T cells via antigen presentation by antigen presenting cells (APCs) along with the action of co-stimulatory molecules and pattern recognition receptors. Cluster of differentiation 40 (CD40) is one such costimulatory molecule that is expressed on APCs that binds to CD40 ligand (CD40L) on T helper cells and activates a signaling cascade, subsequently resulting in a wide range of immune and inflammatory responses. Considering its important role in regulation of immune response, CD40/40 L has been used for developing antitumor vaccines. In this study, we developed methods for evaluating and quantifying the activity of CD40L expressed from an adenovirus vector ONCOS-401. Our results show that the ONCOS-401 vector produces functional CD40L, which can bind and activate a NF-κB-dependent signaling cascade, leading to secreted embryonic alkaline phosphatase reporter production in HEK293-BLUE cells. In addition, quantification of CD40L production using enzyme-linked immunosorbent assay and HEK-293 BLUE reporter cells showed reproducibly higher recovery of CD40L from ONCOS-401 than from the negative control vector or uninfected cells with consistent inter and intra-assay precision. Thus, a rapid and easy method for quantifying and assessing CD40L production and activity from adenovirus vectors would support the assessment of efficacy of the vector for gene therapy - this was the objective of our study.

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Acknowledgements

We thank Antti Vuolanto for the help and assistance with the method development.

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Authors and Affiliations

  1. Targovax Oy, Clinical Science, Helsinki, Finland

    Lukasz Kuryk

  2. Department of Virology, National Institute of Public Health–National Institute of Hygiene, Warsaw, Poland

    Lukasz Kuryk

  3. Targovax ASA, Clinical Science, Oslo, Norway

    Anne-Sophie W. Møller & Magnus Jaderberg

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  1. Lukasz Kuryk
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  2. Anne-Sophie W. Møller
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Correspondence to Lukasz Kuryk.

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LK, ASM, MJ are employees and/or shareholders in Targovax Oy in Finland and Targovax ASA in Norway.

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Kuryk, L., Møller, AS.W. & Jaderberg, M. Quantification and functional evaluation of CD40L production from the adenovirus vector ONCOS-401. Cancer Gene Ther 26, 26–31 (2019). https://doi.org/10.1038/s41417-018-0038-x

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