{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,5]],"date-time":"2026-10-05T12:55:12Z","timestamp":1791204912243,"version":"4.1.0"},"reference-count":87,"publisher":"Wiley","issue":"5","license":[{"start":{"date-parts":[[2026,8,24]],"date-time":"2026-08-24T00:00:00Z","timestamp":1787529600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,8,24]],"date-time":"2026-08-24T00:00:00Z","timestamp":1787529600000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Veterinary Medicine &amp;amp; Sci"],"published-print":{"date-parts":[[2026,9]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:sec>\n                    <jats:title>Background<\/jats:title>\n                    <jats:p>\n                      <jats:italic>Toxoplasma gondii<\/jats:italic>\n                      is a protozoan parasite of medical and veterinary importance causing abortion. While current therapies are limited to address the chronic phase of infection, effective prophylactic vaccines are warranted.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Objectives<\/jats:title>\n                    <jats:p>In this study, we applied a rational vaccine design approach centred around immunodominant dense granule antigens (GRAs).<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>\n                      In silico epitope mapping was performed on six GRAs (GRA15, GRA60, GRA76, GRA83, GRA\u2010\u03b1 and GRA\u2010\u03b2). Using web servers, B\u2010cell, cytotoxic T\u2010lymphocyte and helper T\u2010lymphocyte epitopes were predicted and then filtered for antigenicity, solubility and allergenicity. We designed four vaccine constructs by fusing predicted epitopes with specific linkers and adjuvants (RS\u201009, RpfE\/50S ribosomal protein of\n                      <jats:italic>Mycobacterium tuberculosis<\/jats:italic>\n                      , and human interferon gamma [IFN\u2010\u03b3]). Constructs were subjected to rigorous evaluation, which included physicochemical evaluation, 3D structural prediction and refinement, toll\u2010like receptor\u20104 (TLR\u20104) docking, immune simulation and codon adaptation.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      All candidates exhibited high antigenicity (VaxiJen scores &gt;\u00a00.9) and solubility (&gt;\u00a00.6). Rpf\u2010Toxo emerged optimal, with non\u2010allergenic, antigenic (0.9148), soluble (0.658), stable (instability index: 35.41) and hydrophilic (grand average of hydropathicity: \u22120.641) properties. Docking revealed strong TLR\u20104 binding activity (affinity: \u221220.6\u00a0kcal\/mol;\n                      <jats:italic>K<\/jats:italic>\n                      <jats:sub>d<\/jats:sub>\n                      : 7.3e\u201016\u00a0M) and immune simulation predicted robust responses, including antibody titers &gt;\u00a0170,000, Th1\u2010skewed IFN\u2010\u03b3 (380,000\u00a0ng\/mL) and memory cell activation; however, these in silico predictions need experimental validation. Codon optimization enhanced expression (CAI: 1.00; GC: 65.63%), and\u00a0in silico\u00a0cloning indicated compatibility with pET28a(+).\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusion<\/jats:title>\n                    <jats:p>These computational predictions require future experimental validation through in vitro and in vivo studies to confirm safety and protective efficacy.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1002\/vms3.71189","type":"journal-article","created":{"date-parts":[[2026,8,25]],"date-time":"2026-08-25T04:26:52Z","timestamp":1787632012000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Rpf\u2010Toxo: A Preliminary Computationally Designed Dense Granule Antigen\u2010Based Multi\u2010Epitope Vaccine Against\n                    <i>Toxoplasma 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