{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,2]],"date-time":"2026-06-02T13:19:51Z","timestamp":1780406391261,"version":"3.54.1"},"reference-count":44,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2026,2,5]],"date-time":"2026-02-05T00:00:00Z","timestamp":1770249600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100004442","name":"Narodowym Centrum Nauki","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100004442","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Immunol."],"abstract":"<jats:sec>\n                    <jats:title>Introduction<\/jats:title>\n                    <jats:p>\n                      <jats:italic>Toxoplasma gondii<\/jats:italic>\n                      invasion relies on the battery of specialized proteins, such as ROP8, which plays a key role in formation of parasitophorous vacuoles within host cells. The aim of the work was to evaluate diagnostic, immunoprophylactic and therapeutic potential of recombinant ROP8 (rROP8) protein as despite many years of research this parasitosis still is considered a significant medical and economic problem and the main areas requiring improvement are the diagnostics of the infection, vaccine development especially for humans and safe, specific chemotherapy.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>\n                      Recombinant ROP8 protein (rROP8) of\n                      <jats:italic>Toxoplasma gondii<\/jats:italic>\n                      was cloned, expressed in\n                      <jats:italic>E. coli<\/jats:italic>\n                      Rosetta(DE3)pLysS, and purified by nickel-affinity chromatography. Its diagnostic potential was evaluated using ELISA on sera from experimentally infected mice and human patients. The immunogenic properties of rROP8 were assessed through\n                      <jats:italic>in vitro<\/jats:italic>\n                      stimulation of human monocytes, mouse macrophages, and monocyte-derived dendritic cells, followed by cytokine profiling and co-culture with autologous CD4\n                      <jats:sup>+<\/jats:sup>\n                      lymphocytes. Finally, drug affinity responsive target stability (DARTS) assays and confocal microscopy were used to investigate the interaction of thiazolidin-4-one derivatives with rROP8 and their inhibitory effects on parasitophorous vacuole formation.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      The ROP8 proved to be promising as a potential diagnostic tool enabling the detection of specific IgG antibodies in mouse and human immune sera comparably to the native parasite antigen (TLA). The protein was also capable of activating human monocytes and mouse macrophages\n                      <jats:italic>in vitro<\/jats:italic>\n                      leading to the enhanced clearance of\n                      <jats:italic>T. gondii<\/jats:italic>\n                      tachyzoites via phagocytosis. The protein was also able to activate human monocyte-derived dendritic cells as shown by the cytometric analysis of surface markers and multiplex detection of specific cytokines, also in co-cultures with homologous lymphocytes, leading to Th1\/Th17 lymphocyte phenotype. Finally, the proteolytic degradation of rROP8 was significantly inhibited by the presence of all thiazolidin-4-one derivatives, which exhibited high activity against\n                      <jats:italic>T. gondii<\/jats:italic>\n                      in previous studies.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.3389\/fimmu.2026.1768856","type":"journal-article","created":{"date-parts":[[2026,2,5]],"date-time":"2026-02-05T06:44:49Z","timestamp":1770273889000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":1,"title":["Recombinant ROP8 antigen: diagnostics, immunogenicity, and therapeutic targeting in toxoplasmosis"],"prefix":"10.3389","volume":"17","author":[{"given":"Maciej","family":"Chyb","sequence":"first","affiliation":[{"name":"Department of Molecular Microbiology, Faculty of Biology and Environmental Protection, University of Lodz","place":["Lodz, Poland"]},{"name":"The Bio-Med-Chem Doctoral School of the University of Lodz and Lodz Institutes of the Polish Academy of Sciences, Faculty of Biology and Environmental Protection, University of Lodz","place":["Lodz, Poland"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Adrian","family":"Bekier","sequence":"additional","affiliation":[{"name":"Department of Molecular Microbiology, Faculty of Biology and Environmental Protection, University of Lodz","place":["Lodz, Poland"]},{"name":"Department of Microbiology and Laboratory Medical Immunology, Medical University of Lodz","place":["Lodz, Poland"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Malwina","family":"Kawka","sequence":"additional","affiliation":[{"name":"Department of Molecular Microbiology, Faculty of Biology and Environmental Protection, University of Lodz","place":["Lodz, Poland"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bart\u0142omiej","family":"Ferra","sequence":"additional","affiliation":[{"name":"Department of Tropical Parasitology, Institute of Maritime and Tropical Medicine in Gdynia, Medical University of Gda\u0144sk","place":["Gdynia, Poland"]},{"name":"Institute of Molecular Biology and Biotechnology (IMBB), Foundation for Research and Technology \u2013 Hellas (FORTH)","place":["Heraklion, Greece"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bo\u017cena","family":"Dziadek","sequence":"additional","affiliation":[{"name":"Department of Molecular Microbiology, Faculty of Biology and Environmental Protection, University of Lodz","place":["Lodz, Poland"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sylwia","family":"Michlewska","sequence":"additional","affiliation":[{"name":"Laboratory of Microscopic Imaging and Specialized Biological Techniques, Faculty of Biology and Environmental Protection, University of Lodz","place":["Lodz, Poland"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Katarzyna","family":"Dzitko","sequence":"additional","affiliation":[{"name":"Department of Molecular Microbiology, Faculty of Biology and Environmental Protection, University of Lodz","place":["Lodz, Poland"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nazar","family":"Trotsko","sequence":"additional","affiliation":[{"name":"Department of Organic Chemistry, Faculty of Pharmacy, Medical University of Lublin","place":["Lublin, Poland"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Justyna","family":"Gatkowska","sequence":"additional","affiliation":[{"name":"Department of Molecular Microbiology, Faculty of Biology and Environmental Protection, University of Lodz","place":["Lodz, Poland"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2026,2,5]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","DOI":"10.1128\/CMR.05013-11","article-title":"Epidemiology of and diagnostic strategies for toxoplasmosis","volume":"25","author":"Robert-Gangneux","year":"2012","journal-title":"Clin Microbiol Rev"},{"key":"B2","doi-asserted-by":"publisher","DOI":"10.4067\/s0718-04622007000200009","article-title":"Toxoplasmosis","volume":"363","author":"Eriz","year":"2007","journal-title":"Atenea"},{"key":"B3","doi-asserted-by":"publisher","DOI":"10.1017\/S0031182021001013","article-title":"Congenital toxoplasmosis in humans: An update of worldwide rate of congenital infections","volume":"148","author":"Dubey","year":"2021","journal-title":"Parasitology"},{"key":"B4","doi-asserted-by":"publisher","DOI":"10.1016\/j.pt.2020.08.005","article-title":"Toxoplasma gondii: an underestimated threat","volume":"36","author":"Milne","year":"2020","journal-title":"Trends Parasitol"},{"key":"B5","doi-asserted-by":"publisher","first-page":"e00037","DOI":"10.1016\/j.fawpar.2019.e00037","article-title":"Toxoplasma gondii infection and toxoplasmosis in farm animals: Risk factors and economic impact","volume":"15","author":"Stelzer","year":"2019","journal-title":"Food Waterborne Parasitol"},{"key":"B6","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1186\/s13071-020-04445-z","article-title":"The life-cycle of Toxoplasma gondii reviewed using animations","volume":"13","author":"Attias","year":"2020","journal-title":"Parasit Vectors"},{"key":"B7","article-title":"Overview of apoptosis, autophagy, and inflammatory processes in toxoplasma gondii infected cells","volume-title":"Pathogens.","author":"Ahmadpour","year":"2023"},{"key":"B8","doi-asserted-by":"publisher","first-page":"4","DOI":"10.7774\/cevr.2019.8.1.4","article-title":"Rhoptry antigens as Toxoplasma gondii vaccine target","volume":"8","author":"Foroutan","year":"2019","journal-title":"Clin Exp Vaccine Res"},{"key":"B9","doi-asserted-by":"publisher","first-page":"79","DOI":"10.1038\/nrmicro1800","article-title":"Kiss and spit: The dual roles of Toxoplasma rhoptries","volume":"6","author":"Boothroyd","year":"2008","journal-title":"Nat Rev Microbiol"},{"key":"B10","doi-asserted-by":"publisher","DOI":"10.1016\/j.mib.2007.09.013","article-title":"Rhoptries: an arsenal of secreted virulence factors","volume":"10","author":"Bradley","year":"2007","journal-title":"Curr\u00a0Opin Microbiol"},{"key":"B11","doi-asserted-by":"publisher","first-page":"193","DOI":"10.1016\/j.meegid.2018.04.033","article-title":"Bioinformatics analysis of ROP8 protein to improve vaccine design against Toxoplasma gondii","volume":"62","author":"Foroutan","year":"2018","journal-title":"Infection Genet Evol"},{"key":"B12","doi-asserted-by":"publisher","first-page":"297","DOI":"10.1186\/1756-3305-7-297","article-title":"Sero-diagnostic evaluation of Toxoplasma gondii recombinant Rhoptry antigen 8 expressed in E","volume":"7","author":"Sonaimuthu","year":"2014","journal-title":"coli. Parasit Vectors"},{"key":"B13","doi-asserted-by":"publisher","first-page":"101413","DOI":"10.1016\/j.cimid.2020.101413","article-title":"Vaccination with a novel multi-epitope ROP8 DNA vaccine against acute Toxoplasma gondii infection induces strong B and T cell responses in mice","volume":"69","author":"Foroutan","year":"2020","journal-title":"Comp Immunol Microbiol Infect Dis"},{"key":"B14","doi-asserted-by":"publisher","DOI":"10.4269\/ajtmh.12-0727","article-title":"Protective immune response in BALB\/c mice induced by DNA vaccine of the ROP8 gene of Toxoplasma gondii","volume":"88","author":"Parthasarathy","year":"2013","journal-title":"Am J Trop Med Hygiene"},{"key":"B15","doi-asserted-by":"publisher","DOI":"10.1007\/s11224-019-01458-y","article-title":"In silico studies of novel scaffold of thiazolidin-4-one derivatives as anti-Toxoplasma gondii agents by 2D\/3D-QSAR, molecular docking, and molecular dynamics simulations","volume":"31","author":"Abdizadeh","year":"2020","journal-title":"Struct Chem"},{"key":"B16","doi-asserted-by":"publisher","DOI":"10.3390\/ijms25084384","article-title":"The development of toxoplasma gondii recombinant trivalent chimeric proteins as an alternative to toxoplasma lysate antigen (TLA) in enzyme-linked immunosorbent assay (ELISA) for the detection of immunoglobulin G (IgG) in small ruminants","volume":"25","author":"Ferra","year":"2024","journal-title":"Int J Mol Sci"},{"key":"B17","first-page":"e3000411","article-title":"Reporting animal research: Explanation and elaboration for the arrive guidelines 2.0","volume-title":"PLoS Biology.","author":"du Sert","year":"2020"},{"key":"B18","doi-asserted-by":"publisher","first-page":"5621","DOI":"10.3390\/ijms26125621","article-title":"The trivalent\u00a0recombinant chimeric proteins containing immunodominant fragments\u00a0of toxoplasma gondii sag1\u00a0and sag2 antigens in their core\u2014a good diagnostic tool for detecting igg antibodies in human serum samples","volume":"26","author":"Ferra","year":"2025","journal-title":"Int J Mol Sci"},{"key":"B19","doi-asserted-by":"publisher","first-page":"12976","DOI":"10.1038\/s41598-023-40147-z","article-title":"Evaluation of long-term immunity and protection against T. gondii after immunization with multivalent recombinant chimeric T. gondii proteins","volume":"13","author":"Chyb","year":"2023","journal-title":"Sci Rep"},{"key":"B20","doi-asserted-by":"publisher","first-page":"812","DOI":"10.3390\/molecules22050812","article-title":"Evaluation of the mycobactericidal effect of thio-functionalized carbohydrate derivatives","volume":"22","author":"Korycka-Machala","year":"2017","journal-title":"Molecules"},{"key":"B21","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2022.833635","article-title":"Halophilic archaea halorhabdus\u00a0rudnickae and natrinema salaciae activate human dendritic cells and orient T helper cell responses","volume":"13","author":"Krawczyk","year":"2022","journal-title":"Front Immunol"},{"key":"B22","doi-asserted-by":"publisher","first-page":"3029","DOI":"10.3390\/molecules24173029","article-title":"Synthesis and in vitro anti-Toxoplasma gondii activity of novel thiazolidin-4-one derivatives","volume":"24","author":"Trotsko","year":"2019","journal-title":"Molecules"},{"key":"B23","doi-asserted-by":"publisher","first-page":"3213","DOI":"10.3390\/ijms23063213","article-title":"4-arylthiosemicarbazide derivatives as toxoplasmic aromatic amino acid hydroxylase inhibitors and anti-inflammatory agents","volume":"23","author":"Bekier","year":"2022","journal-title":"Int J Mol Sci"},{"key":"B24","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2024.1385582","article-title":"Identification of novel biomarkers for anti-Toxoplasma gondii IgM detection and the potential application in rapid diagnostic fluorescent tests","volume":"15","author":"Nguyen","year":"2024","journal-title":"Front Microbiol"},{"key":"B25","doi-asserted-by":"publisher","DOI":"10.1017\/S0031182009991636","article-title":"Ovine toxoplasmosis","volume":"136","author":"Innes","year":"2010","journal-title":"Parasitology"},{"key":"B26","doi-asserted-by":"publisher","first-page":"131","DOI":"10.1038\/s41541-022-00563-0","article-title":"Toxoplasmosis vaccines: what we have and where to go","volume":"7","author":"Zhang","year":"2022","journal-title":"NPJ Vaccines"},{"key":"B27","doi-asserted-by":"publisher","DOI":"10.1016\/j.pt.2019.01.005","article-title":"Advances in the development of anti-toxoplasma gondii vaccines: challenges, opportunities, and perspectives","volume":"35","author":"Wang","year":"2019","journal-title":"Trends Parasitol"},{"key":"B28","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2018.02587","article-title":"Drug resistance in Toxoplasma gondii","volume":"9","author":"Montazeri","year":"2018","journal-title":"Front Microbiol"},{"key":"B29","doi-asserted-by":"publisher","first-page":"43","DOI":"10.3390\/pathogens9010043","article-title":"Toxoplasma gondii recombinant antigen AMA1: Diagnostic utility of protein fragments for the detection of IgG and IgM antibodies","volume":"9","author":"Ferra","year":"2020","journal-title":"Pathogens"},{"key":"B30","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2024.1356216","article-title":"IFNs in host defence and parasite immune evasion during Toxoplasma gondii infections","volume":"15","author":"L\u00fcder","year":"2024","journal-title":"Front Immunol"},{"key":"B31","doi-asserted-by":"publisher","first-page":"2082","DOI":"10.3390\/vaccines","article-title":"Exploiting the Macrophage Production of IL-12 in Improvement of Vaccine Development against Toxoplasma gondii and Neospora caninum Infections","volume":"10","author":"Massari","year":"2022"},{"key":"B32","doi-asserted-by":"crossref","DOI":"10.1007\/978-1-0716-1888-2_8","article-title":"Macrophage Stimulation as a Useful Approach for Immunoscreening of Potential Vaccine Candidates Against Toxoplasma gondii and Neospora caninum Infections","volume-title":"Methods Mol Biol","author":"Fereig","year":"2022"},{"key":"B33","doi-asserted-by":"publisher","DOI":"10.1038\/sj.onc.1209943","article-title":"NF-\u03baB and the immune response","volume":"25","author":"Hayden","year":"2006","journal-title":"Oncogene"},{"key":"B34","doi-asserted-by":"publisher","first-page":"106650","DOI":"10.1016\/j.actatropica.2022.106650","article-title":"Toxoplasma gondii rhoptry protein (TgROP18) enhances the expression of pro-inflammatory factor in LPS\/IFN-\u03b3-induced murine BV2 microglia cells via NF-\u03baB signal pathway","volume":"235","author":"Xue","year":"2022","journal-title":"Acta Trop"},{"key":"B35","doi-asserted-by":"publisher","first-page":"154","DOI":"10.3390\/vaccines7040154","article-title":"The impact of the antigenic composition of chimeric proteins on their immunoprotective activity against chronic toxoplasmosis in mice","volume":"7","author":"Gatkowska","year":"2019","journal-title":"Vaccines (Basel)"},{"key":"B36","doi-asserted-by":"publisher","DOI":"10.3389\/fcell.2019.00091","article-title":"TNFR1 and TNFR2 in the control of the life and death balance of macrophages","volume":"7","author":"Wajant","year":"2019","journal-title":"Front Cell Dev Biol"},{"key":"B37","doi-asserted-by":"publisher","first-page":"e100279","DOI":"10.1371\/journal.ppat.1002794","article-title":"Phagocyte responses to protozoan infection and how toxoplasma gondii meets the challenge","volume":"8","author":"Denkers","year":"2012","journal-title":"PloS Pathog"},{"key":"B38","volume-title":"Monitoring dendritic cell activation and maturation.,\u201d Methods in Molecular Biology","author":"Macri","year":"2019"},{"key":"B39","doi-asserted-by":"publisher","first-page":"925135","DOI":"10.1155\/2012\/925135","article-title":"CD4 +T cells: Differentiation and functions","volume":"2012","author":"Luckheeram","year":"2012","journal-title":"Clin Dev Immunol"},{"key":"B40","doi-asserted-by":"publisher","first-page":"e12532","DOI":"10.1111\/pim.12532","article-title":"T-cell profiles elicited by Toxoplasma gondii in acutely\/chronically infected humans","volume":"40","author":"Silva-Gutierrez","year":"2018","journal-title":"Parasite Immunol"},{"key":"B41","doi-asserted-by":"publisher","first-page":"38","DOI":"10.3390\/ijms24010038","article-title":"Memory macrophages","volume":"24","author":"Kloc","year":"2023","journal-title":"Int J Mol Sci"},{"key":"B42","doi-asserted-by":"publisher","first-page":"79","DOI":"10.1007\/s00281-009-0191-2","article-title":"Th17 cytokines and vaccine-induced immunity","volume":"32","author":"Lin","year":"2010","journal-title":"Semin Immunopathol"},{"key":"B43","doi-asserted-by":"publisher","first-page":"105150","DOI":"10.1016\/j.meegid.2021.105150","article-title":"Evaluation of Th17 immune responses of recombinant DNA vaccine encoding GRA14 and ROP13 genes against Toxoplasma gondii in BALB\/c mice","volume":"96","author":"Fatollahzadeh","year":"2021","journal-title":"Infection Genet Evol"},{"key":"B44","doi-asserted-by":"publisher","first-page":"e1005902","DOI":"10.1371\/journal.ppat.1005902","article-title":"Th17 cells are more protective than th1 cells against the intracellular parasite trypanosoma cruzi","volume":"12","author":"Cai","year":"2016","journal-title":"PloS Pathog"}],"container-title":["Frontiers in Immunology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fimmu.2026.1768856\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,2,5]],"date-time":"2026-02-05T06:44:51Z","timestamp":1770273891000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fimmu.2026.1768856\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,2,5]]},"references-count":44,"alternative-id":["10.3389\/fimmu.2026.1768856"],"URL":"https:\/\/doi.org\/10.3389\/fimmu.2026.1768856","relation":{},"ISSN":["1664-3224"],"issn-type":[{"value":"1664-3224","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,2,5]]},"article-number":"1768856"}}