Jansons, J. | Search bioRxiv

bioRxiv2026

A plant virus-based virus like-particle platform for therapeutic vaccination targeting IL-23A

Interleukin-23 (IL-23) is a pivotal cytokine in the pathogenesis of immune-mediated inflammatory diseases, making it an attractive therapeutic target. Vaccination against self-antigens offers an alternative to monoclonal antibody (mAb) therapies by eliciting endogenous Ab responses. Here, we report the development of a virus-like particle (VLP)-based vaccine targeting murine IL-23A (mIL23A) using the eggplant mosaic virus (EMV) coat protein as a carrier. mIL23A was incorporated either as a direct fusion or within a mosaic system, with the latter supporting efficient VLP assembly. Alternative antigen presentation strategies employed synthetic coiled-coil and zipper pairs. All constructs were expressed in E. coli, purified, and characterized by SDS-PAGE, TEM, and DLS. Immunization of BALB/c mice elicited robust mIL23A-specific Abs, with mosaic VLPs inducing high-avidity, Th1-biased responses. Encapsidated RNA within VLPs was double-stranded and stimulated TLR3, enhancing immunogenicity. Scale-up in bioreactors preserved VLP integrity and antigen incorporation, yielding 1.72 mg per gram of cells. These results establish EMV VLPs as a flexible and scalable platform for presenting challenging antigens and provide a promising approach for therapeutic vaccination against self-antigens.

immunology↗

bioRxiv2022

Identification and full genome analysis of the first putative virus of sea buckthorn (Hippophae rhamnoides L.)

The agricultural importance of sea buckthorn (Hippophae rhamnoides L.) is rapidly increasing. Several bacterial and fungal pathogens infecting sea buckthorn have been identified and characterized; however, the viral pathogens are not yet known. In this study, we identified, isolated, and sequenced a virus from a wild plantation of sea buckthorn for the first time. Sequence analysis of the obtained viral genome revealed high similarity with sequences of several viruses belonging to the genus Marafivirus, especially olive latent virus 3 (OLV-3). The genome of the new virus is 6,989 nucleotides (nt) in length according to 5' and 3' rapid amplification of cDNA ends (RACE) (without polyA-tail), with 5' and 3' untranslated regions being 133 and 109 nt long, respectively. The viral genome encoded two open reading frames (ORFs). ORF1 encoded a polyprotein of 1,954 amino acids (aa) with the characteristic marafivirus non-structural protein domains--methyltransferase, Salyut domain, papain-like cysteine protease, helicase, and RNA-dependent RNA polymerase. ORF1 was separated from ORF2 by a six nt and encoded the coat protein (CP). CP had typical signatures of minor (30.96 kDa) and major (21.18 kDa) forms. Both CP forms were cloned and expressed in a bacterial expression system, and only the major CP was able to self-assemble into 30 nm virus-like particles that resembled the native virus, thus demonstrating that minor CP is not essential for virion assembly. We suggest the newly discovered virus to be named as "Sea buckthorn marafivirus", abbreviated as "SBuMV". Author summarySea buckthorn is an exceptionally valuable plant that is currently widely cultivated as multipurpose horticultural species for food, pharmacology, cosmetics, and landscape conservation. Diseases and pests directly affect the cultivation of SBT. To date, several pests and diseases, mainly fungal and bacterial, but no viral, sea buckthorn have been reported. Identification of new pathogens would assist in the development of control strategies, and quarantine purposes and ensure sustainable sea buckthorn cultivation. Here, for the first time, we present a virus putatively infecting sea buckthorn. We had characterized its full genome, cloned and expressed minor and major forms of coat protein either individually or co-expressed. We also showed that major coat protein-derived virus-like particles self- assembled directly in the bacterial cells, and the majority of the expressed CPs were soluble. Our study suggests that the minor CP is not essential for the assembly of seemingly structurally intact viral particles, meaning that it can have other functions.

molecular biology↗