{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,2]],"date-time":"2026-09-02T09:24:13Z","timestamp":1788341053177,"version":"build-2803163510"},"reference-count":50,"publisher":"MDPI AG","issue":"9","license":[{"start":{"date-parts":[[2026,9,2]],"date-time":"2026-09-02T00:00:00Z","timestamp":1788307200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"FBIS SSC VB \u201cVector\u201d Rospotrebnadzor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Vaccines"],"abstract":"<jats:p>Background\/Objectives: In recent years, researchers have directed considerable attention toward the activation of the T-cell response in the development of vaccines against viral infections, including the tick-borne encephalitis virus (TBEV). A particularly promising approach to developing effective and safe T-cell vaccines involves the use of artificial multi-epitope immunogens. The selection of spacers that link epitopes within a construct can have a significant effect on the immunogenicity of multi-epitope constructs. The objective of this study was to design multi-epitope TBEV immunogens using various spacers and to evaluate their immunogenicity. Methods: The present study involved the design of three multi-epitope immunogens, which were developed based on T-cell epitopes from TBEV proteins. The AG1-ub construct contained optimized alanine spacers, the AG2-ub construct contained GPGPG spacers, and the AG4-ub construct contained no spacers. Three DNA vaccines encoding the designed immunogens were subsequently produced. To assess the immunogenicity of the constructs, BALB\/c mice were immunized with the designed DNA vaccines via electroporation. Results: The ELISpot assay demonstrated that DNA vaccines encoding AG1-ub and AG4-ub induced a significant number of IFN-\u03b3-producing cells. The DNA vaccine encoding AG2-ub, a multi-epitope with GPGPG spacers, exhibited low immunogenicity, potentially attributable to inadequate processing or misfolding of the AG2-ub protein, thereby affecting the molecule\u2019s structure. Conclusions: The data provided in this study on the effect of spacers on the immunogenicity of multi-epitope constructs can be used to optimize the design of vaccine candidates. However, further research is needed to assess their clinical potential.<\/jats:p>","DOI":"10.3390\/vaccines14090770","type":"journal-article","created":{"date-parts":[[2026,9,2]],"date-time":"2026-09-02T08:56:01Z","timestamp":1788339361000},"page":"770","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["How Does Inter-Epitope Spacer Variation Within Artificial Immunogens Based on T-Cell Epitopes of Tick-Borne Encephalitis Virus Affect Immunogenicity?"],"prefix":"10.3390","volume":"14","author":[{"given":"Elena V.","family":"Yakovleva","sequence":"first","affiliation":[{"name":"State Scientific Center of Virology and Biotechnology \u201cVector\u201d, Rospotrebnadzor, 630559 Koltsovo, 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Scientific Center of Virology and Biotechnology \u201cVector\u201d, Rospotrebnadzor, 630559 Koltsovo, Russia"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"methodology","vocabulary":"credit"},{"role":"investigation","vocabulary":"credit"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1733-9524","authenticated-orcid":false,"given":"Ekaterina V.","family":"Starostina","sequence":"additional","affiliation":[{"name":"State Scientific Center of Virology and Biotechnology \u201cVector\u201d, Rospotrebnadzor, 630559 Koltsovo, Russia"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"methodology","vocabulary":"credit"},{"role":"investigation","vocabulary":"credit"}]},{"given":"Vladimir A.","family":"Yakovlev","sequence":"additional","affiliation":[{"name":"State Scientific Center of Virology and Biotechnology \u201cVector\u201d, Rospotrebnadzor, 630559 Koltsovo, 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630559 Koltsovo, Russia"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"methodology","vocabulary":"credit"},{"role":"investigation","vocabulary":"credit"}]},{"given":"Andrey P.","family":"Rudometov","sequence":"additional","affiliation":[{"name":"State Scientific Center of Virology and Biotechnology \u201cVector\u201d, Rospotrebnadzor, 630559 Koltsovo, Russia"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"conceptualization","vocabulary":"credit"},{"role":"validation","vocabulary":"credit"},{"role":"writing-review-editing","vocabulary":"credit"},{"role":"supervision","vocabulary":"credit"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4365-8809","authenticated-orcid":false,"given":"Larisa I.","family":"Karpenko","sequence":"additional","affiliation":[{"name":"State Scientific Center of Virology and Biotechnology \u201cVector\u201d, Rospotrebnadzor, 630559 Koltsovo, 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