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Article

In Vitro Evaluation of Glycoengineered RSV-F in the Human Artificial Lymph Node Reactor

1
Molecular Biotechnology and Functional Genomics, Technical University of Applied Sciences Wildau, Hochschulring 1, Wildau 15745, Germany
2
Institute of Pathology, Charitè-University Medicine Berlin, Augustenburger Platz 1, Berlin 13353, Germany
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Laboratory of Biochemistry, Department of Life Sciences and Technology, Beuth University of Applied Sciences, Seestraße 64, Berlin 13347, Germany
4
ProBioGen AG, Goethestraße 54, Berlin 13086, Germany
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Department of Life Science Engineering, HTW Berlin University of Applied Sciences, Wilhelminenhofstraße 75a, Berlin 12459, Germany
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Institute of Laboratory Medicine, Clinical Chemistry and Pathobiochemistry, Charité Medical University Berlin, Augustenburger Platz 1, Berlin 13353, Germany
*
Author to whom correspondence should be addressed.
Academic Editor: Anthony Guiseppi-Elie
Bioengineering 2017, 4(3), 70; https://doi.org/10.3390/bioengineering4030070
Received: 1 April 2017 / Revised: 27 July 2017 / Accepted: 3 August 2017 / Published: 15 August 2017
(This article belongs to the Special Issue Recombinant Glycoproteins)
Subunit vaccines often require adjuvants to elicit sustained immune activity. Here, a method is described to evaluate the efficacy of single vaccine candidates in the preclinical stage based on cytokine and gene expression analysis. As a model, the recombinant human respiratory syncytial virus (RSV) fusion protein (RSV-F) was produced in CHO cells. For comparison, wild-type and glycoengineered, afucosylated RSV-F were established. Both glycoprotein vaccines were tested in a commercial Human Artificial Lymph Node in vitro model (HuALN®). The analysis of six key cytokines in cell culture supernatants showed well-balanced immune responses for the afucosylated RSV-F, while immune response of wild-type RSV-F was more Th1 accentuated. In particular, stronger and specific secretion of interleukin-4 after each round of re-stimulation underlined higher potency and efficacy of the afucosylated vaccine candidate. Comprehensive gene expression analysis by nCounter gene expression assay confirmed the stronger onset of the immunologic reaction in stimulation experiments with the afucosylated vaccine in comparison to wild-type RSV-F and particularly revealed prominent activation of Th17 related genes, innate immunity, and comprehensive activation of humoral immunity. We, therefore, show that our method is suited to distinguish the potency of two vaccine candidates with minor structural differences. View Full-Text
Keywords: Glycoengineering; fucosylation; RSV; F-Protein; NanoString Glycoengineering; fucosylation; RSV; F-Protein; NanoString
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MDPI and ACS Style

Radke, L.; Sandig, G.; Lubitz, A.; Schließer, U.; Von Horsten, H.H.; Blanchard, V.; Keil, K.; Sandig, V.; Giese, C.; Hummel, M.; Hinderlich, S.; Frohme, M. In Vitro Evaluation of Glycoengineered RSV-F in the Human Artificial Lymph Node Reactor. Bioengineering 2017, 4, 70. https://doi.org/10.3390/bioengineering4030070

AMA Style

Radke L, Sandig G, Lubitz A, Schließer U, Von Horsten HH, Blanchard V, Keil K, Sandig V, Giese C, Hummel M, Hinderlich S, Frohme M. In Vitro Evaluation of Glycoengineered RSV-F in the Human Artificial Lymph Node Reactor. Bioengineering. 2017; 4(3):70. https://doi.org/10.3390/bioengineering4030070

Chicago/Turabian Style

Radke, Lars, Grit Sandig, Annika Lubitz, Ulrike Schließer, Hans H. Von Horsten, Veronique Blanchard, Karolin Keil, Volker Sandig, Christoph Giese, Michael Hummel, Stephan Hinderlich, and Marcus Frohme. 2017. "In Vitro Evaluation of Glycoengineered RSV-F in the Human Artificial Lymph Node Reactor" Bioengineering 4, no. 3: 70. https://doi.org/10.3390/bioengineering4030070

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