{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,21]],"date-time":"2026-01-21T10:36:46Z","timestamp":1768991806222,"version":"3.49.0"},"reference-count":29,"publisher":"MDPI AG","issue":"2","license":[{"start":{"date-parts":[[2023,2,5]],"date-time":"2023-02-05T00:00:00Z","timestamp":1675555200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001871","name":"Funda\u00e7\u00e3o para a Ci\u00eancia e a Tecnologia","doi-asserted-by":"publisher","award":["PTDC\/CVT\/01234\/2008"],"award-info":[{"award-number":["PTDC\/CVT\/01234\/2008"]}],"id":[{"id":"10.13039\/501100001871","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001871","name":"Funda\u00e7\u00e3o para a Ci\u00eancia e a Tecnologia","doi-asserted-by":"publisher","award":["311931"],"award-info":[{"award-number":["311931"]}],"id":[{"id":"10.13039\/501100001871","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001871","name":"Funda\u00e7\u00e3o para a Ci\u00eancia e a Tecnologia","doi-asserted-by":"publisher","award":["UIDB\/00276\/2020"],"award-info":[{"award-number":["UIDB\/00276\/2020"]}],"id":[{"id":"10.13039\/501100001871","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001871","name":"Funda\u00e7\u00e3o para a Ci\u00eancia e a Tecnologia","doi-asserted-by":"publisher","award":["LA\/P\/0059\/2020"],"award-info":[{"award-number":["LA\/P\/0059\/2020"]}],"id":[{"id":"10.13039\/501100001871","id-type":"DOI","asserted-by":"publisher"}]},{"name":"European Union\u05f3s Seventh Framework Programme","award":["PTDC\/CVT\/01234\/2008"],"award-info":[{"award-number":["PTDC\/CVT\/01234\/2008"]}]},{"name":"European Union\u05f3s Seventh Framework Programme","award":["311931"],"award-info":[{"award-number":["311931"]}]},{"name":"European Union\u05f3s Seventh Framework Programme","award":["UIDB\/00276\/2020"],"award-info":[{"award-number":["UIDB\/00276\/2020"]}]},{"name":"European Union\u05f3s Seventh Framework Programme","award":["LA\/P\/0059\/2020"],"award-info":[{"award-number":["LA\/P\/0059\/2020"]}]},{"DOI":"10.13039\/501100001871","name":"Funda\u00e7\u00e3o para a Ci\u00eancia e a Tecnologia","doi-asserted-by":"publisher","award":["PTDC\/CVT\/01234\/2008"],"award-info":[{"award-number":["PTDC\/CVT\/01234\/2008"]}],"id":[{"id":"10.13039\/501100001871","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001871","name":"Funda\u00e7\u00e3o para a Ci\u00eancia e a Tecnologia","doi-asserted-by":"publisher","award":["311931"],"award-info":[{"award-number":["311931"]}],"id":[{"id":"10.13039\/501100001871","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001871","name":"Funda\u00e7\u00e3o para a Ci\u00eancia e a Tecnologia","doi-asserted-by":"publisher","award":["UIDB\/00276\/2020"],"award-info":[{"award-number":["UIDB\/00276\/2020"]}],"id":[{"id":"10.13039\/501100001871","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001871","name":"Funda\u00e7\u00e3o para a Ci\u00eancia e a Tecnologia","doi-asserted-by":"publisher","award":["LA\/P\/0059\/2020"],"award-info":[{"award-number":["LA\/P\/0059\/2020"]}],"id":[{"id":"10.13039\/501100001871","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001871","name":"AL4AnimalS","doi-asserted-by":"publisher","award":["PTDC\/CVT\/01234\/2008"],"award-info":[{"award-number":["PTDC\/CVT\/01234\/2008"]}],"id":[{"id":"10.13039\/501100001871","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001871","name":"AL4AnimalS","doi-asserted-by":"publisher","award":["311931"],"award-info":[{"award-number":["311931"]}],"id":[{"id":"10.13039\/501100001871","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001871","name":"AL4AnimalS","doi-asserted-by":"publisher","award":["UIDB\/00276\/2020"],"award-info":[{"award-number":["UIDB\/00276\/2020"]}],"id":[{"id":"10.13039\/501100001871","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001871","name":"AL4AnimalS","doi-asserted-by":"publisher","award":["LA\/P\/0059\/2020"],"award-info":[{"award-number":["LA\/P\/0059\/2020"]}],"id":[{"id":"10.13039\/501100001871","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Viruses"],"abstract":"<jats:p>The African Swine Fever Virus (ASFV) is an economically important, large DNA virus which causes a highly contagious and frequently fatal disease in domestic pigs. Due to the acute nature of the infection and the complexity of the protective porcine anti-ASFV response, there is no accepted vaccine in use. As resistance to ASFV is known to correlate with a robust IFN response, the virus is predicted to have evolved strategies to inhibit innate immunity by modulating the IFN response. The deletion of virus host evasion gene(s) inhibiting IFN is a logical solution to develop an attenuated virus vaccine. One such candidate, the ASFV ORF I329L gene, is highly conserved in pathogenic and non-pathogenic virus isolates and in this study we confirm and extend the conclusion that it has evolved for the inhibition of innate immunity initiated through Toll-like receptors (TLRs). Specifically, the ASFV I329L extracellular (ECD) and intracellular (ICD) domains inhibit TLR signalling by two entirely different mechanisms. Bioinformatics modelling suggests that the ECD inhibits several TLR signalling pathways through a short sequence homologous to the conserved TLR dimerization domain, here termed the putative dimerization domain (PDD). Remarkably, both full length and PDD constructs of I329L were demonstrated to inhibit activation, not only of TLR3, but also TLR4, TLR5, TLR8 and TLR9. Additionally, the demonstration of a weak association of I329L with TLR3 is consistent with the formation of a non-signalling I329L-TLR3 heterodimer, perhaps mediated through the PDD of I329L. Finally, the ICD associates with TRIF, thereby impacting on both TLR3 and TLR4 signalling. Thus, I329L offers potential as a general inhibitor of TLR responses and is a rational candidate for construction and testing of an I329L deletion mutant vaccine.<\/jats:p>","DOI":"10.3390\/v15020445","type":"journal-article","created":{"date-parts":[[2023,2,6]],"date-time":"2023-02-06T04:08:07Z","timestamp":1675656487000},"page":"445","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":14,"title":["I329L: A Dual Action Viral Antagonist of TLR Activation Encoded by the African Swine Fever Virus (ASFV)"],"prefix":"10.3390","volume":"15","author":[{"given":"S\u00edlvia","family":"Correia","sequence":"first","affiliation":[{"name":"Instituto Gulbenkian de Ci\u00eancia, 2780-156 Oeiras, Portugal"},{"name":"CIISA\u2014Centro de Investiga\u00e7\u00e3o Interdisciplinar em Sanidade Animal, Faculdade de Medicina Veterin\u00e1ria, Universidade de Lisboa, 1300-477 Lisbon, Portugal"},{"name":"Laborat\u00f3rio Associado para Ci\u00eancia Animal e Veterin\u00e1ria (AL4AnimalS), 1300-477 Lisbon, Portugal"}]},{"given":"Pedro Lu\u00eds","family":"Moura","sequence":"additional","affiliation":[{"name":"Instituto Gulbenkian de Ci\u00eancia, 2780-156 Oeiras, Portugal"}]},{"given":"S\u00f3nia","family":"Ventura","sequence":"additional","affiliation":[{"name":"Instituto Gulbenkian de Ci\u00eancia, 2780-156 Oeiras, Portugal"}]},{"given":"Alexandre","family":"Leit\u00e3o","sequence":"additional","affiliation":[{"name":"CIISA\u2014Centro de Investiga\u00e7\u00e3o Interdisciplinar em Sanidade Animal, Faculdade de Medicina Veterin\u00e1ria, Universidade de Lisboa, 1300-477 Lisbon, Portugal"},{"name":"Laborat\u00f3rio Associado para Ci\u00eancia Animal e Veterin\u00e1ria (AL4AnimalS), 1300-477 Lisbon, Portugal"}]},{"given":"Robert Michael Evans","family":"Parkhouse","sequence":"additional","affiliation":[{"name":"Instituto Gulbenkian de Ci\u00eancia, 2780-156 Oeiras, Portugal"}]}],"member":"1968","published-online":{"date-parts":[[2023,2,5]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Van Etten, J.L. (2009). Lesser Known Large dsDNA Viruses, Springer. Current Topics in Microbiology and Immunology.","DOI":"10.1007\/978-3-540-68618-7"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"e1179","DOI":"10.1111\/tbed.14466","article-title":"One Hundred Years of African Swine Fever in Africa: Where Have We Been, Where Are We Now, Where Are We Going?","volume":"69","author":"Penrith","year":"2022","journal-title":"Transbound. Emerg. Dis."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"87","DOI":"10.1016\/j.virusres.2012.10.013","article-title":"Identification and Utility of Innate Immune System Evasion Mechanisms of ASFV","volume":"173","author":"Correia","year":"2013","journal-title":"Virus Res."},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Zhu, J.J. (2022). African Swine Fever Vaccinology: The Biological Challenges from Immunological Perspectives. Viruses, 14.","DOI":"10.3390\/v14092021"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"10514","DOI":"10.1128\/JVI.00485-06","article-title":"Macrophage Transcriptional Responses Following In Vitro Infection with a Highly Virulent African Swine Fever Virus Isolate","volume":"80","author":"Zhang","year":"2006","journal-title":"J. Virol."},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"He, W.-R., Yuan, J., Ma, Y.-H., Zhao, C.-Y., Yang, Z.-Y., Zhang, Y., Han, S., Wan, B., and Zhang, G.-P. (2022). Modulation of Host Antiviral Innate Immunity by African Swine Fever Virus: A Review. Animals, 12.","DOI":"10.3390\/ani12212935"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"597","DOI":"10.1007\/s00705-010-0894-7","article-title":"A Novel TLR3 Inhibitor Encoded by African Swine Fever Virus (ASFV)","volume":"156","author":"Almeida","year":"2011","journal-title":"Arch. Virol."},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Reis, A.L., Goatley, L.C., Jabbar, T., Lopez, E., Rathakrishnan, A., and Dixon, L.K. (2020). Deletion of the Gene for the Type I Interferon Inhibitor I329L from the Attenuated African Swine Fever Virus OURT88\/3 Strain Reduces Protection Induced in Pigs. Vaccines, 8.","DOI":"10.3390\/vaccines8020262"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"247","DOI":"10.1002\/pro.554","article-title":"Modeling of the Toll-like Receptor 3 and a Putative Toll-like Receptor 3 Antagonist Encoded by the African Swine Fever Virus","volume":"20","author":"Henriques","year":"2011","journal-title":"Protein Sci."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"363","DOI":"10.1038\/nprot.2009.2","article-title":"Protein Structure Prediction on the Web: A Case Study Using the Phyre Server","volume":"4","author":"Kelley","year":"2009","journal-title":"Nat. Protoc."},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Zhang, Y. (2008). I-TASSER Server for Protein 3D Structure Prediction. BMC Bioinform., 9.","DOI":"10.1186\/1471-2105-9-40"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"2.9.1","DOI":"10.1002\/0471140864.ps0209s50","article-title":"Comparative Protein Structure Modeling Using MODELLER","volume":"50","author":"Eswar","year":"2007","journal-title":"Curr. Protoc. Protein Sci."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"283","DOI":"10.1107\/S0021889892009944","article-title":"PROCHECK: A Program to Check the Stereochemical Quality of Protein Structures","volume":"26","author":"Laskowski","year":"1993","journal-title":"J. Appl. Crystallogr."},{"key":"ref_14","first-page":"932","article-title":"Gene Transduction Efficiency in Cells of Different Species by HIV and EIAV Vectors","volume":"9","author":"Ikeda","year":"2002","journal-title":"Gene"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"379","DOI":"10.1126\/science.1155406","article-title":"Structural Basis of Toll-Like Receptor 3 Signaling with Double-Stranded RNA","volume":"320","author":"Liu","year":"2008","journal-title":"Science"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"761","DOI":"10.1038\/nsmb.1453","article-title":"A Second Binding Site for Double-Stranded RNA in TLR3 and Consequences for Interferon Activation","volume":"15","author":"Pirher","year":"2008","journal-title":"Nat. Struct. Mol. Biol."},{"key":"ref_17","first-page":"298","article-title":"Three B-Cell Surface Molecules Associating with Membrane Immunoglobulin","volume":"69","author":"Parkhouse","year":"1990","journal-title":"Immunology"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"13","DOI":"10.1007\/s11262-022-01939-z","article-title":"Attenuated African Swine Fever Virus through Serial Passaging of Viruses in Cell Culture: A Brief Review on the Knowledge Gathered during 60 Years of Research","volume":"59","author":"Zhang","year":"2022","journal-title":"Virus Genes"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"2694","DOI":"10.1016\/j.vaccine.2018.03.040","article-title":"African Swine Fever Virus (ASFV) Protection Mediated by NH\/P68 and NH\/P68 Recombinant Live-Attenuated Viruses","volume":"36","author":"Gallardo","year":"2018","journal-title":"Vaccine"},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Gladue, D.P., O\u2019Donnell, V., Ramirez-Medina, E., Rai, A., Pruitt, S., Vuono, E.A., Silva, E., Velazquez-Salinas, L., and Borca, M.V. (2020). Deletion of CD2-Like (CD2v) and C-Type Lectin-Like (EP153R) Genes from African Swine Fever Virus Georgia-\u22069GL Abrogates Its Effectiveness as an Experimental Vaccine. Viruses, 12.","DOI":"10.3390\/v12101185"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"449","DOI":"10.1016\/j.it.2012.05.002","article-title":"Dissecting Negative Regulation of Toll-like Receptor Signaling","volume":"33","author":"Kondo","year":"2012","journal-title":"Trends Immunol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"301","DOI":"10.3389\/fimmu.2018.00301","article-title":"Molecular Signatures of a TLR4 Agonist-Adjuvanted HIV-1 Vaccine Candidate in Humans","volume":"9","author":"Anderson","year":"2018","journal-title":"Front. Immunol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"510","DOI":"10.1016\/j.coph.2012.06.002","article-title":"New Developments in Toll-like Receptor Targeted Therapeutics","volume":"12","author":"Connolly","year":"2012","journal-title":"Curr. Opin. Pharmacol."},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Szabo, A., Gogolak, P., Pazmandi, K., Kis-Toth, K., Riedl, K., Wizel, B., Lingnau, K., Bacsi, A., Rethi, B., and Rajnavolgyi, E. (2013). The Two-Component Adjuvant IC31\u00ae Boosts Type I Interferon Production of Human Monocyte-Derived Dendritic Cells via Ligation of Endosomal TLRs. PLoS ONE, 8.","DOI":"10.1371\/annotation\/322893f7-942c-4059-9295-7ecd26162aca"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"9","DOI":"10.1016\/j.cellimm.2010.10.008","article-title":"Novel Antagonist Antibody to TLR3 Blocks Poly(I:C)-Induced Inflammation in Vivo and in Vitro","volume":"267","author":"Bunting","year":"2011","journal-title":"Cell. Immunol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"778","DOI":"10.1128\/CMR.14.4.778-809.2001","article-title":"Antiviral Actions of Interferons","volume":"14","author":"Samuel","year":"2001","journal-title":"Clin. Microbiol. Rev."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1099\/vir.0.83391-0","article-title":"Interferons and Viruses: An Interplay between Induction, Signalling, Antiviral Responses and Virus Countermeasures","volume":"89","author":"Randall","year":"2008","journal-title":"J. Gen. Virol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"1858","DOI":"10.1128\/JVI.78.4.1858-1864.2004","article-title":"African Swine Fever Virus Multigene Family 360 and 530 Genes Affect Host Interferon Response","volume":"78","author":"Afonso","year":"2004","journal-title":"J. Virol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"198931","DOI":"10.1016\/j.virusres.2022.198931","article-title":"African Swine Fever Virus MGF505-7R Protein Interacted with IRF7and TBK1 to Inhibit Type I Interferon Production","volume":"322","author":"Yang","year":"2022","journal-title":"Virus Res."}],"container-title":["Viruses"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1999-4915\/15\/2\/445\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T18:25:03Z","timestamp":1760120703000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1999-4915\/15\/2\/445"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,2,5]]},"references-count":29,"journal-issue":{"issue":"2","published-online":{"date-parts":[[2023,2]]}},"alternative-id":["v15020445"],"URL":"https:\/\/doi.org\/10.3390\/v15020445","relation":{},"ISSN":["1999-4915"],"issn-type":[{"value":"1999-4915","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,2,5]]}}}