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

Title:
In Vitro Evaluation of Glycoengineered RSV-F in the Human Artificial Lymph Node Reactor
Abstract:
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.
Year:
2017
Publication type:
Journal article
Journal:
Bioengineering
ISSN:
2306-5354
Number:
3
Volume:
4
Pages:
70
Language:
English
Document status:
Open access
PubListerURL:
https://publister.bib.th-wildau.de/publister/public/publication/2344
Keywords:
Glycoengineering; fucosylation; RSV; F-Protein; NanoString
Radke, L., Sandig, G., Lubitz, A., Schließer, U., von Horsten, H., Blanchard, V., et al. (2017). In Vitro Evaluation of Glycoengineered RSV-F in the Human Artificial Lymph Node Reactor Bioengineering. 4 (3), 70.
@article{2344,
    author           = {Radke, Lars and Sandig, Grit and Lubitz, Annika and Schließer, Ulrike and von Horsten, Hans Henning and Blanchard, Véronique and Keil, Karolin  and Sandig, Volker and Giese, Christoph and Hummel, Michael and Hinderlich, Stephan and Frohme, Marcus},
    title            = {In Vitro Evaluation of Glycoengineered RSV-F in the Human Artificial Lymph Node Reactor},
    journal          = {Bioengineering},
    year             = {2017},
    volume           = {4},
    number           = {3},
    pages            = {70},
    publisher        = {MDPI},
    doi              = {10.3390/bioengineering4030070},
}


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