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.
Highlights
In recent years, the process of vaccine development has evolved drastically
We focused on the highly-conserved respiratory syncytial virus (RSV)-F which is responsible for virus spreading by fusing with neighboring healthy cells
It is more likely that differently-processed proteins exist. Both furin cleavage sites are obviously recognized by the protease, part of RSV-F might only be processed by one furin cleavage and still contain the pep27 peptide, whereas the lower band seems to represent the fully-processed F2 subunit after double furin cleavage
Summary
The process of vaccine development has evolved drastically. Yet, the main requirement on vaccines have not changed: the establishment of a long-lasting and pathogen-specific immunological memory as a result of a concerted immune response. Recombinant protein vaccines opened a way to introduce immunogenic agents immediately during the production process This could be done by additional immunogenic peptides, which are bound N- or C-terminally to the protein backbone, or by introduction of non-human posttranslational modifications, e.g., via glycosylation in the case of glycoprotein vaccines [4]. These foreign glycosidic structures are able to interact with the conserved Toll-like receptors and, by that, stimulate the immune system comprehensively. This is in contrast to monosaccharides artificial to humans, e.g., N-glycan-bound xylose, which may work as adjuvants [4,5,6]
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
