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The viral protein 1 (VP1) contains major antigenic and neutralizing epitopes, making it a promising target for subunit vaccine development.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Aims<\/jats:title>\n                    <jats:p>This study evaluated the immunogenicity of a recombinant VP1 fragment (amino acids 198-297) fused to a truncated Newcastle Disease Virus nucleoprotein (NPt-VP1t).<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Materials and methodology<\/jats:title>\n                    <jats:p>\n                      Soluble NPt-VP1t (SP) and insoluble NPt-VP1t (IB) versions of the protein were expressed in\n                      <jats:italic>E. coli<\/jats:italic>\n                      , purified, and verified by SDS-PAGE and Western blot using anti-VP1 and anti-NDV antibodies, confirming the ~59 kDa fusion protein. Adult female BALB\/c mice were immunized intraperitoneally with SP, IB, or PBS control, with two booster doses at two-week intervals.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      Results demonstrated that mice immunized with the SP formulation produced significantly higher anti-VP1 IgG reactivity than those receiving the IB or controls (\n                      <jats:italic>p<\/jats:italic>\n                      \u2009&lt;\u20090.05). After the first booster (week 4), the antibody level in the SP was approximately 2-fold higher than the IB, with the highest OD readings observed at week 8 post-immunization. The SP maintained high antibody levels for at least two weeks post-booster. Splenocyte proliferation assays revealed that the SB group had a stimulation index (S.I.) of 0.826\u2009\u00b1\u20090.104, about 1.6 times greater than the IB group (\n                      <jats:italic>p<\/jats:italic>\n                      \u2009&lt;\u20090.05). Cytokine profiling showed significantly elevated Th1 (IFN-\u03b3, IL-2) and Th2 (IL-4, IL-6, IL-10) cytokines in both SP and IB groups compared to controls (\n                      <jats:italic>p<\/jats:italic>\n                      \u2009&lt;\u20090.05), with IFN-\u03b3 levels markedly higher in vaccinated mice, indicating activation of both humoral and cell-mediated immunity. SDS-PAGE of the IB protein revealed contaminant bands, suggesting the actual amount of NPt-VP1t administered was lower than in the soluble formulation, potentially affecting its immunogenicity. All groups received formulations with Freund\u2019s adjuvant, which may limit assessment of vaccine-specific safety.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusion<\/jats:title>\n                    <jats:p>Overall, the SP recombinant NPt-VP1t protein elicited robust humoral and cellular immune responses in mice, outperforming the insoluble form. These findings support the immunogenic potential of SP as a subunit vaccine candidate for EV71. Future studies should include viral challenge and neutralization assays to confirm protective efficacy.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1007\/s42770-025-01784-w","type":"journal-article","created":{"date-parts":[[2025,9,23]],"date-time":"2025-09-23T08:13:15Z","timestamp":1758615195000},"page":"2743-2755","update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Soluble recombinant enterovirus 71 VP1 fused to truncated newcastle disease virus nucleoprotein elicits immune responses in mice"],"prefix":"10.1007","volume":"56","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-3384-1397","authenticated-orcid":false,"given":"Suhaili","family":"Mustafa","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Noraini","family":"Abd-Aziz","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Khatijah","family":"Yusoff","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Norazizah","family":"Shafee","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2025,9,23]]},"reference":[{"issue":"1","key":"1784_CR1","doi-asserted-by":"publisher","first-page":"169","DOI":"10.3390\/ijms24010169","volume":"24","author":"X Zhang","year":"2023","unstructured":"Zhang X, Zhang Y, Li H, Liu L (2023) Hand-Foot-and-Mouth Disease-Associated enterovirus and the development of multivalent HFMD vaccines. 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