{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,6]],"date-time":"2026-09-06T04:49:52Z","timestamp":1788670192036,"version":"build-2803163510"},"reference-count":53,"publisher":"MDPI AG","issue":"1","license":[{"start":{"date-parts":[[2023,12,26]],"date-time":"2023-12-26T00:00:00Z","timestamp":1703548800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"university\u2013industry cooperation project of the Fujian 793 Province of China","award":["2021N5003"],"award-info":[{"award-number":["2021N5003"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>African swine fever (ASF) leads to high mortality in domestic pigs and wild boar, and it is caused by the African swine fever virus (ASFV). Currently, no commercially available vaccine exists for its prevention in China. In this study, we engineered a pseudorabies recombinant virus (PRV) expressing ASFV CD2v and p54 proteins (PRV-\u2206TK-(CD2v)-\u2206gE-(p54)) using CRISPR\/Cas9 and homologous recombination technology. PRV-\u2206TK-(CD2v)-\u2206gE-(p54) effectively delivers CD2v and p54, and it exhibits reduced virulence. Immunization with PRV-\u2206TK-(CD2v)-\u2206gE-(p54) neither induces pruritus nor causes systemic infection and inflammation. Furthermore, a double knockout of the TK and gE genes eliminates the depletion of T, B, and monocytes\/macrophages in the blood caused by wild-type viral infection, decreases the proliferation of granulocytes to eliminate T-cell immunosuppression from granulocytes, and enhances the ability of the immune system against PRV infection. An overexpression of CD2v and p54 proteins does not alter the characteristics of PRV-\u2206TK\/\u2206gE. Moreover, PRV-\u2206TK-(CD2v)-\u2206gE-(p54) successfully induces antibody production via intramuscular (IM) vaccination and confers effective protection for vaccinated mice upon challenge. Thus, PRV-\u2206TK-(CD2v)-\u2206gE-(p54) demonstrates good immunogenicity and safety, providing highly effective protection against PRV and ASFV. It potentially represents a suitable candidate for the development of a bivalent vaccine against both PRV and ASFV infections.<\/jats:p>","DOI":"10.3390\/ijms25010335","type":"journal-article","created":{"date-parts":[[2023,12,26]],"date-time":"2023-12-26T04:40:44Z","timestamp":1703565644000},"page":"335","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":11,"title":["Generation and Characterization of Recombinant Pseudorabies Virus Delivering African Swine Fever Virus CD2v and p54"],"prefix":"10.3390","volume":"25","author":[{"given":"Jianhui","family":"Wei","sequence":"first","affiliation":[{"name":"Fujian Key Laboratory of Innate Immune Biology, Biomedical Research Center of South China, College of Life Science, Fujian Normal University Qishan Campus, Fuzhou 350117, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0007-9317-1798","authenticated-orcid":false,"given":"Chuancheng","family":"Liu","sequence":"additional","affiliation":[{"name":"Fujian Key Laboratory of Innate Immune Biology, Biomedical Research Center of South China, College of Life Science, Fujian Normal University Qishan Campus, Fuzhou 350117, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xinyan","family":"He","sequence":"additional","affiliation":[{"name":"Fujian Key Laboratory of Innate Immune Biology, Biomedical Research Center of South China, College of Life Science, Fujian Normal University Qishan Campus, Fuzhou 350117, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bilal","family":"Abbas","sequence":"additional","affiliation":[{"name":"Fujian Key Laboratory of Innate Immune Biology, Biomedical Research Center of South China, College of Life Science, Fujian Normal University Qishan Campus, Fuzhou 350117, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9063-6819","authenticated-orcid":false,"given":"Qi","family":"Chen","sequence":"additional","affiliation":[{"name":"Fujian Key Laboratory of Innate Immune Biology, Biomedical Research Center of South China, College of Life Science, Fujian Normal University Qishan Campus, Fuzhou 350117, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhaolong","family":"Li","sequence":"additional","affiliation":[{"name":"Institute of Animal Husbandry and Veterinary Medicine, Fujian Academy of Agricultural Sciences, Fuzhou 350117, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3465-645X","authenticated-orcid":false,"given":"Zhihua","family":"Feng","sequence":"additional","affiliation":[{"name":"Fujian Key Laboratory of Innate Immune Biology, Biomedical Research Center of South China, College of Life Science, Fujian Normal University Qishan Campus, Fuzhou 350117, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2023,12,26]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"157","DOI":"10.1016\/j.virs.2022.03.006","article-title":"Regulation of antiviral immune response by African swine fever virus (ASFV)","volume":"37","author":"Zheng","year":"2022","journal-title":"Virol. Sin."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"715582","DOI":"10.3389\/fimmu.2021.715582","article-title":"Structure of African Swine Fever Virus and Associated Molecular Mechanisms Underlying Infection and Immunosuppression: A Review","volume":"12","author":"Wang","year":"2021","journal-title":"Front. Immunol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"613","DOI":"10.1099\/jgv.0.001049","article-title":"ICTV Virus Taxonomy Profile: Asfarviridae","volume":"99","author":"Alonso","year":"2018","journal-title":"J. Gen. Virol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"249","DOI":"10.1006\/viro.1995.1149","article-title":"Analysis of the complete nucleotide sequence of African swine fever virus","volume":"208","author":"Nogal","year":"1995","journal-title":"Virology"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"3","DOI":"10.1016\/j.virusres.2012.10.020","article-title":"African swine fever virus replication and genomics","volume":"173","author":"Dixon","year":"2013","journal-title":"Virus Res."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"146","DOI":"10.1038\/s41392-022-00996-y","article-title":"COVID-19 vaccine development: Milestones, lessons and prospects","volume":"7","author":"Li","year":"2022","journal-title":"Signal Transduct. Target. Ther."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"749","DOI":"10.1016\/S2213-2600(22)00131-X","article-title":"Safety and immunogenicity of a live-attenuated influenza virus vector-based intranasal SARS-CoV-2 vaccine in adults: Randomised, double-blind, placebo-controlled, phase 1 and 2 trials","volume":"10","author":"Zhu","year":"2022","journal-title":"Lancet Respir. Med."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"e2102775118","DOI":"10.1073\/pnas.2102775118","article-title":"Scalable live-attenuated SARS-CoV-2 vaccine candidate demonstrates preclinical safety and efficacy","volume":"118","author":"Wang","year":"2021","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"109493","DOI":"10.1016\/j.celrep.2021.109493","article-title":"Development of safe and highly protective live-attenuated SARS-CoV-2 vaccine candidates by genome recoding","volume":"36","author":"Trimpert","year":"2021","journal-title":"Cell Rep."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"623","DOI":"10.1007\/s11427-020-1657-9","article-title":"A seven-gene-deleted African swine fever virus is safe and effective as a live attenuated vaccine in pigs","volume":"63","author":"Chen","year":"2020","journal-title":"Sci. China Life Sci."},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Borca, M.V., Ramirez-Medina, E., Silva, E., Vuono, E., Rai, A., Pruitt, S., Holinka, L.G., Velazquez-Salinas, L., Zhu, J., and Gladue, D.P. (2020). Development of a Highly Effective African Swine Fever Virus Vaccine by Deletion of the I177L Gene Results in Sterile Immunity against the Current Epidemic Eurasia Strain. J. Virol., 94.","DOI":"10.1128\/JVI.02017-19"},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Teklue, T., Wang, T., Luo, Y., Hu, R., Sun, Y., and Qiu, H.J. (2020). Generation and Evaluation of an African Swine Fever Virus Mutant with Deletion of the CD2v and UK Genes. Vaccines, 8.","DOI":"10.3390\/vaccines8040763"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"4593","DOI":"10.1016\/j.vaccine.2011.04.052","article-title":"Protection of European domestic pigs from virulent African isolates of African swine fever virus by experimental immunisation","volume":"29","author":"King","year":"2011","journal-title":"Vaccine"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"2905","DOI":"10.1080\/21645515.2020.1820808","article-title":"Recombinant vaccines for COVID-19","volume":"16","author":"Yadav","year":"2020","journal-title":"Hum. Vaccin. Immunother."},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Zhou, M., Abid, M., Cao, S., and Zhu, S. (2023). Recombinant Pseudorabies Virus Usage in Vaccine Development against Swine Infectious Disease. Viruses, 15.","DOI":"10.3390\/v15020370"},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Jin, Y.L., Yin, D., Xing, G., Huang, Y.M., Fan, C.M., Fan, C.F., Qiu, X.H., Dong, W.R., Yan, Y., and Gu, J.Y. (2022). The Inactivated gE\/TK Gene-Deleted Vaccine Against Pseudorabies Virus Type II Confers Effective Protection in Mice and Pigs. Front. Microbiol., 13.","DOI":"10.3389\/fmicb.2022.943707"},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Boadella, M., Gort\u00e1zar, C., Vicente, J., and Ruiz-Fons, F. (2012). Wild boar: An increasing concern for Aujeszky\u2019s disease control in pigs?. BMC Vet. Res., 8.","DOI":"10.1186\/1746-6148-8-7"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"462","DOI":"10.1128\/MMBR.69.3.462-500.2005","article-title":"Molecular biology of pseudorabies virus: Impact on neurovirology and veterinary medicine","volume":"69","author":"Pomeranz","year":"2005","journal-title":"Microbiol. Mol. Biol. Rev."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"102","DOI":"10.3201\/eid2001.130531","article-title":"Pathogenic pseudorabies virus, China, 2012","volume":"20","author":"Yu","year":"2014","journal-title":"Emerg. Infect. Dis."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"1749","DOI":"10.3201\/eid1911.130177","article-title":"Pseudorabies virus variant in Bartha-K61-vaccinated pigs, China, 2012","volume":"19","author":"An","year":"2013","journal-title":"Emerg. Infect. Dis."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"23","DOI":"10.5501\/wjv.v5.i1.23","article-title":"Pathogenicity of a currently circulating Chinese variant pseudorabies virus in pigs","volume":"5","author":"Yang","year":"2016","journal-title":"World J. Virol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"272","DOI":"10.1186\/1743-422X-8-272","article-title":"Growth, physicochemical properties, and morphogenesis of Chinese wild-type PRV Fa and its gene-deleted mutant strain PRV SA215","volume":"8","author":"Zhu","year":"2011","journal-title":"Virol. J."},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Li, W., Zhuang, D., Li, H., Zhao, M., Zhu, E., Xie, B., Chen, J., and Zhao, M. (2021). Recombinant pseudorabies virus with gI\/gE deletion generated by overlapping polymerase chain reaction and homologous recombination technology induces protection against the PRV variant PRV-GD2013. BMC Vet. Res., 17.","DOI":"10.1186\/s12917-021-02861-6"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"797611","DOI":"10.3389\/fvets.2021.797611","article-title":"Construction and Immunogenicity of a Recombinant Pseudorabies Virus Variant With TK\/gI\/gE\/11k\/28k Deletion","volume":"8","author":"Yan","year":"2021","journal-title":"Front. Vet. Sci."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"424","DOI":"10.1128\/JVI.78.1.424-440.2004","article-title":"Complete, annotated sequence of the pseudorabies virus genome","volume":"78","author":"Klupp","year":"2004","journal-title":"J. Virol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"3294","DOI":"10.1128\/JVI.01994-10","article-title":"Processing and localization of the african swine fever virus CD2v transmembrane protein","volume":"85","author":"Goatley","year":"2011","journal-title":"J. Virol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"5312","DOI":"10.1128\/jvi.67.9.5312-5320.1993","article-title":"African swine fever virus encodes a CD2 homolog responsible for the adhesion of erythrocytes to infected cells","volume":"67","author":"Rodriguez","year":"1993","journal-title":"J. Virol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"2881","DOI":"10.1128\/JVI.72.4.2881-2889.1998","article-title":"Deletion of a CD2-like gene, 8-DR, from African swine fever virus affects viral infection in domestic swine","volume":"72","author":"Borca","year":"1998","journal-title":"J. Virol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"285","DOI":"10.1006\/viro.1996.0193","article-title":"Functional and immunological properties of the baculovirus-expressed hemagglutinin of African swine fever virus","volume":"218","author":"Escribano","year":"1996","journal-title":"Virology"},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"Hua, R.H., Liu, J., Zhang, S.J., Liu, R.Q., Zhang, X.F., He, X.J., Zhao, D.M., and Bu, Z.G. (2023). Mammalian Cell-Line-Expressed CD2v Protein of African Swine Fever Virus Provides Partial Protection against the HLJ\/18 Strain in the Early Infection Stage. Viruses, 15.","DOI":"10.3390\/v15071467"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"2265661","DOI":"10.1080\/22221751.2023.2265661","article-title":"A protective multiple gene-deleted African swine fever virus genotype II, Georgia 2007\/1, expressing a modified non-haemadsorbing CD2v protein","volume":"12","author":"Rathakrishnan","year":"2023","journal-title":"Emerg. Microbes Infect."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"180","DOI":"10.1186\/s12985-020-01450-7","article-title":"The recombinant pseudorabies virus expressing African swine fever virus CD2v protein is safe and effective in mice","volume":"17","author":"Feng","year":"2020","journal-title":"Virol. J."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"224","DOI":"10.1016\/j.febslet.2004.06.001","article-title":"The African swine fever virus dynein-binding protein p54 induces infected cell apoptosis","volume":"569","author":"Dixon","year":"2004","journal-title":"FEBS Lett."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"461","DOI":"10.1006\/viro.1998.9068","article-title":"The African swine fever virus proteins p54 and p30 are involved in two distinct steps of virus attachment and both contribute to the antibody-mediated protective immune response","volume":"243","author":"Oviedo","year":"1998","journal-title":"Virology"},{"key":"ref_35","doi-asserted-by":"crossref","unstructured":"Laval, K., Vernejoul, J.B., Van Cleemput, J., Koyuncu, O.O., and Enquist, L.W. (2018). Virulent Pseudorabies Virus Infection Induces a Specific and Lethal Systemic Inflammatory Response in Mice. J. Virol., 92.","DOI":"10.1128\/JVI.01614-18"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"6048","DOI":"10.1128\/JVI.00554-15","article-title":"African Swine Fever Virus Georgia Isolate Harboring Deletions of MGF360 and MGF505 Genes Is Attenuated in Swine and Confers Protection against Challenge with Virulent Parental Virus","volume":"89","author":"Holinka","year":"2015","journal-title":"J. Virol."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"8556","DOI":"10.1128\/JVI.00969-15","article-title":"African Swine Fever Virus Georgia 2007 with a Deletion of Virulence-Associated Gene 9GL (B119L), when Administered at Low Doses, Leads to Virus Attenuation in Swine and Induces an Effective Protection against Homologous Challenge","volume":"89","author":"Holinka","year":"2015","journal-title":"J. Virol."},{"key":"ref_38","doi-asserted-by":"crossref","unstructured":"Monteagudo, P.L., Lacasta, A., L\u00f3pez, E., Bosch, L., Collado, J., Pina-Pedrero, S., Correa-Fiz, F., Accensi, F., Navas, M.J., and Vidal, E. (2017). BA71\u0394CD2: A New Recombinant Live Attenuated African Swine Fever Virus with Cross-Protective Capabilities. J. Virol., 91.","DOI":"10.1128\/JVI.01058-17"},{"key":"ref_39","doi-asserted-by":"crossref","unstructured":"Wang, J., Song, Z., Ge, A., Guo, R., Qiao, Y., Xu, M., Wang, Z., Liu, Y., Zheng, Y., and Fan, H. (2018). Safety and immunogenicity of an attenuated Chinese pseudorabies variant by dual deletion of TK&gE genes. BMC Vet. Res., 14.","DOI":"10.1186\/s12917-018-1536-7"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"824630","DOI":"10.1155\/2014\/824630","article-title":"An overview of live attenuated recombinant pseudorabies viruses for use as novel vaccines","volume":"2014","author":"Dong","year":"2014","journal-title":"J. Immunol. Res."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"307","DOI":"10.1186\/1743-422X-8-307","article-title":"A novel recombinant pseudorabies virus expressing parvovirus VP2 gene: Immunogenicity and protective efficacy in swine","volume":"8","author":"Chen","year":"2011","journal-title":"Virol. J."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"3963","DOI":"10.1128\/MCB.05297-11","article-title":"Phosphotyrosine-dependent coupling of Tim-3 to T-cell receptor signaling pathways","volume":"31","author":"Lee","year":"2011","journal-title":"Mol. Cell. Biol."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"411","DOI":"10.1093\/intimm\/14.4.411","article-title":"IFN-alpha\/beta enhances BCR-dependent B cell responses","volume":"14","author":"Braun","year":"2002","journal-title":"Int. Immunol."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"294","DOI":"10.1089\/vim.2019.0207","article-title":"B Cell Activation and Response Regulation During Viral Infections","volume":"33","author":"Lam","year":"2020","journal-title":"Viral. Immunol."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"4343","DOI":"10.4049\/jimmunol.176.7.4343","article-title":"Type I IFN receptor signals directly stimulate local B cells early following influenza virus infection","volume":"176","author":"Coro","year":"2006","journal-title":"J. Immunol."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"2094","DOI":"10.1002\/eji.200635993","article-title":"Early type I interferon-mediated signals on B cells specifically enhance antiviral humoral responses","volume":"36","author":"Fink","year":"2006","journal-title":"Eur. J. Immunol."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"355","DOI":"10.1038\/nature02284","article-title":"Lymphocyte egress from thymus and peripheral lymphoid organs is dependent on S1P receptor 1","volume":"427","author":"Matloubian","year":"2004","journal-title":"Nature"},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"1693","DOI":"10.3389\/fimmu.2017.01693","article-title":"Role of Monocyte\/Macrophages during HIV\/SIV Infection in Adult and Pediatric Acquired Immune Deficiency Syndrome","volume":"8","author":"Merino","year":"2017","journal-title":"Front. Immunol."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"723","DOI":"10.1038\/nri3073","article-title":"Protective and pathogenic functions of macrophage subsets","volume":"11","author":"Murray","year":"2011","journal-title":"Nat. Rev. Immunol."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"508","DOI":"10.1111\/j.1365-2141.2005.05794.x","article-title":"Circulating CD10-\/CD16low neutrophils provide a quantitative index of active bone marrow neutrophil release","volume":"131","author":"Orr","year":"2005","journal-title":"Br. J. Haematol."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"887","DOI":"10.1002\/cyto.b.21739","article-title":"Immature Granulocytes: A Risk Factor of Infection after Cardiac Surgery","volume":"94","author":"Daix","year":"2018","journal-title":"Cytom. B Clin. Cytom."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"e0188922","DOI":"10.1128\/jvi.01889-22","article-title":"African Swine Fever Virus Envelope Glycoprotein CD2v Interacts with Host CSF2RA to Regulate the JAK2-STAT3 Pathway and Inhibit Apoptosis to Facilitate Virus Replication","volume":"97","author":"Gao","year":"2023","journal-title":"J. Virol."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"1055","DOI":"10.1128\/CVI.00131-10","article-title":"Correlates of protection induced by vaccination","volume":"17","author":"Plotkin","year":"2010","journal-title":"Clin. Vaccine Immunol."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/25\/1\/335\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T21:42:08Z","timestamp":1760132528000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/25\/1\/335"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,12,26]]},"references-count":53,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2024,1]]}},"alternative-id":["ijms25010335"],"URL":"https:\/\/doi.org\/10.3390\/ijms25010335","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,12,26]]}}}