# Virus-like and Virus Replicon Particles Targeting Multiple B-Cell Antigens Do Not Protect Against African Swine Fever Virus

**Authors:** Kirill Lotonin, Obdulio García-Nicolás, Normann Kilb, Stefan Krämer, Xinyue Chang, Paul Engeroff, Kemal Mehinagic, Noelle Donzé, Francisco Brito, Matthias Liniger, Ilva Lieknina, Darja Cernova, Ieva Balta, Gabriela González-García, Paloma Rueda, Gert Zimmer, Charaf Benarafa, Nicolas Ruggli, Günter Roth, Kaspars Tars, Martin Bachmann, Artur Summerfield

PMC · DOI: 10.3390/vaccines14030285 · Vaccines · 2026-03-23

## TL;DR

Researchers tested virus-like and replicon particles targeting B-cell antigens in pigs but found they did not protect against African swine fever virus.

## Contribution

The study identifies challenges in using B-cell antigens for vaccine development and highlights the importance of cellular immunity in protection against ASFV.

## Key findings

- VLP- and VSV-based vaccines induced antibody responses but failed to provide protection against ASFV.
- Only the attenuated ASFV strain elicited robust T-cell immunity and protection.
- The findings emphasize the critical role of cellular immunity in combating African swine fever.

## Abstract

Background: African swine fever virus (ASFV) causes a fatal hemorrhagic disease in domestic pigs and wild boars. While live attenuated vaccines (LAVs) provide protection, their use raises safety concerns. Therefore, the aim of the present study was to identify viral B-cell antigens associated with protection and to test their potential using highly immunogenic vaccine delivery platforms. Methods: We employed a microarray of 169 ASFV proteins expressed in a cell-free prokaryotic system to identify immunodominant antigens using sera from immune pigs. Six structural proteins were selected and formulated into AP205 virus-like particles (VLPs). Additionally, replication-defective vesicular stomatitis virus (VSV)-based vaccine candidates expressing glycosylated CD2v and EP153R proteins were generated. Three groups of specific pathogen-free pigs were immunized with either VLP- or VSV-based vaccines and challenged with the virulent ASFV Georgia 2007 strain. Control groups included pigs immunized with the attenuated ASFV Estonia 2014 strain and a naïve group. Results: Most vaccine candidates induced detectable antibody responses against target ASFV proteins. However, neither VLP- nor VSV-based vaccines provided protection, as clinical scores, hematology, cytokine responses, and viremia levels were similar to those in the negative control group. In contrast, only the ASFV Estonia 2014 strain elicited a robust T-cell response and protective immunity. Conclusions: These findings highlight the challenges in identifying protective B-cell antigens of ASFV and emphasize the pivotal role of cellular immunity in mediating protection.

## Linked entities

- **Proteins:** EP153R (lectin-like protein)
- **Diseases:** African swine fever (MONDO:0025377), hemorrhagic disease (MONDO:0002243)

## Full-text entities

- **Genes:** IL1B (interleukin 1 beta) [NCBI Gene 3553] {aka IL-1, IL1-BETA, IL1F2, IL1beta}, MBP (myelin basic protein) [NCBI Gene 4155], O61R (structural protein p12) [NCBI Gene 22220327], DDX17 (DEAD-box helicase 17) [NCBI Gene 10521] {aka P72, RH70}, EP402R (CD2 homolog) [NCBI Gene 22220440], CXCL8 (C-X-C motif chemokine ligand 8) [NCBI Gene 3576] {aka GCP-1, GCP1, IL8, LECT, LUCT, LYNAP}, CD8B (CD8 subunit beta) [NCBI Gene 926] {aka CD8B1, CD8beta, LEU2, LY3, LYT3, Ly-3}, EP153R (lectin-like protein) [NCBI Gene 22220439], IFNA1 (interferon alpha 1) [NCBI Gene 3439] {aka IFL, IFN, IFN-ALPHA, IFN-alphaD, IFNA13, IFNA@}, TNF (tumor necrosis factor) [NCBI Gene 7124] {aka DIF, IMD127, TNF-alpha, TNFA, TNFSF2, TNLG1F}, Dync1h1 (dynein cytoplasmic 1 heavy chain 1) [NCBI Gene 13424] {aka 9930018I23Rik, DHC1, DHC1a, DNCL, Dnchc1, Dnec1}, IL1R1 (interleukin 1 receptor type 1) [NCBI Gene 3554] {aka CD121A, CRMO3, D2S1473, IL-1R-alpha, IL-1RT1, IL1R}, I196L (pI196L) [NCBI Gene 22220372], B263R [NCBI Gene 22220316], B407L [NCBI Gene 22220314], IL1A (interleukin 1 alpha) [NCBI Gene 3552] {aka IL-1 alpha, IL-1A, IL1, IL1-ALPHA, IL1F1}, KP177R (structural protein p22) [NCBI Gene 22220394], H124R [NCBI Gene 22220345], IFNG (interferon gamma) [NCBI Gene 3458] {aka IFG, IFI, IMD69}, CD8A (CD8 subunit alpha) [NCBI Gene 925] {aka CD8, CD8alpha, IMD116, Leu2, p32}, B117L [NCBI Gene 22220313], Pold4 (polymerase (DNA-directed), delta 4) [NCBI Gene 69745] {aka 2410012M21Rik, Polds, p12}, POLE4 (DNA polymerase epsilon 4, accessory subunit) [NCBI Gene 56655] {aka YHHQ1, p12}, CHP1 (calcineurin like EF-hand protein 1) [NCBI Gene 11261] {aka CHP, SLC9A1BP, SPAX9, Sid470p, p22, p24}, CD4 (CD4 molecule) [NCBI Gene 920] {aka CD4mut, IMD79, Leu-3, OKT4D, T4}
- **Diseases:** fever (MESH:D005334), swelling (MESH:D004487), Inflammatory (MESH:D007249), thrombocytopenia (MESH:D013921), lymphocytopenia (MESH:D008231), viremia (MESH:D014766), Infection (MESH:D007239), hemorrhagic disease (MESH:D006470), injury to (MESH:D014947)
- **Chemicals:** Brefeldin A (MESH:D020126), Tween (MESH:D011136), CO2 (MESH:D002245), water (MESH:D014867), aluminum (MESH:D000535), copper (MESH:D003300), N2 (MESH:D009584), lysine (MESH:D008239), H2O2 (MESH:D006861), ethanolamine (MESH:D019856), PBS (MESH:D007854), SDS (MESH:D012967), EDTA (MESH:D004492), succinimide (MESH:C032620), AlexaFluor-488 (MESH:C000711379), biotin (MESH:D001710), Ag (MESH:D012834), H2SO4 (MESH:C033158), TCEP (MESH:C080938), uranyl acetate (MESH:C005460), PVDF (MESH:C024865), AP205 VLP (-), mifepristone (MESH:D015735), agarose (MESH:D012685), carbon (MESH:D002244), PDMS (MESH:C013830), IPTG (MESH:D007544)
- **Species:** Sus scrofa (pig, species) [taxon 9823], Suidae (boars, family) [taxon 9821], Bacteriophage sp. (species) [taxon 38018], Escherichia coli (E. coli, species) [taxon 562], Escherichia coli BL21(DE3) (strain) [taxon 469008], Adenoviridae (family) [taxon 10508], African swine fever virus (no rank) [taxon 10497], Homo sapiens (human, species) [taxon 9606], Mus musculus (house mouse, species) [taxon 10090], Azotobacter chroococcum subsp. isscasi (subspecies) [taxon 2528971], Vesicular stomatitis virus (species) [taxon 11276]
- **Mutations:** C with 0, A240L, E165R, K205R, E183L, A104R, A137R, H171R, I73R, F530S
- **Cell lines:** Expi293 — Homo sapiens (Human), Transformed cell line (CVCL_D615), HEK-293 — Homo sapiens (Human), Transformed cell line (CVCL_0045), B169L — Mus musculus (Mouse), Hybridoma (CVCL_B0NT), BHK-G43 — Mesocricetus auratus (Golden hamster), Spontaneously immortalized cell line (CVCL_1914), AP205 — Homo sapiens (Human), Xeroderma pigmentosum variant type, Transformed cell line (CVCL_2556), CCL- — Mus musculus (Mouse), Undefined cell line type (CVCL_M023), Vero — Chlorocebus sabaeus (Green monkey), Spontaneously immortalized cell line (CVCL_0059)

## Full text

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## Figures

8 figures with captions in the complete paper: https://tomesphere.com/paper/PMC13030806/full.md

## References

62 references — full list in the complete paper: https://tomesphere.com/paper/PMC13030806/full.md

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Source: https://tomesphere.com/paper/PMC13030806