{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,29]],"date-time":"2026-10-29T22:21:22Z","timestamp":1793312482791,"version":"4.5.0"},"reference-count":51,"publisher":"MDPI AG","issue":"4","license":[{"start":{"date-parts":[[2021,4,16]],"date-time":"2021-04-16T00:00:00Z","timestamp":1618531200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"International cooperation project of Hubei province","award":["2018AHB011"],"award-info":[{"award-number":["2018AHB011"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Viruses"],"abstract":"<jats:p>Owing to viral evolution and recombination, emerging pseudorabies virus (PRV) strains have caused unprecedented outbreaks in swine farms even when the pigs were previously vaccinated, which might indicate that traditional vaccines were unable to provide effective protection. The development of safe and efficacious vaccines presents prospects to minimize the clinical signs and eventually eradicate the infection. In this study, we used an emerging PRV strain, HNX, as the parental strain to construct a recombinant PRV with TK\/gE gene deletion and Fms-related tyrosine kinase 3 ligand (Flt3L) expression, named HNX-TK\u2212\/gE\u2212-Flt3L. HNX-TK\u2212\/gE\u2212-Flt3L enhanced the maturation of bone marrow derived dendritic cells (DCs) in vitro. Significantly more activated DCs were detected in HNX-TK\u2212\/gE\u2212-Flt3L-immunized mice compared with those immunized with HNX-TK\u2212\/gE\u2212. Subsequently, a remarkable increase of neutralizing antibodies, gB-specific IgG antibodies, and interferon-gamma (IFN-\u03b3) was observed in mice vaccinated with HNX-TK\u2212\/gE\u2212-Flt3L. In addition, a lower mortality and less histopathological damage were observed in HNX-TK\u2212\/gE\u2212-Flt3L vaccinated mice with upon PRV lethal challenge infection. Taken together, our results revealed the potential of Flt3L as an ideal adjuvant that can activate DCs and enhance protective immune responses and support the further evaluation of HNX-TK\u2212\/gE\u2212-Flt3L as a promising PRV vaccine candidate.<\/jats:p>","DOI":"10.3390\/v13040691","type":"journal-article","created":{"date-parts":[[2021,4,19]],"date-time":"2021-04-19T21:59:49Z","timestamp":1618869589000},"page":"691","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":19,"title":["Recombinant Pseudorabies Virus with TK\/gE Gene Deletion and Flt3L Co-Expression Enhances the Innate and Adaptive Immune Response via Activating Dendritic Cells"],"prefix":"10.3390","volume":"13","author":[{"given":"Lun","family":"Yao","sequence":"first","affiliation":[{"name":"State Key Laboratory of Agricultural Microbiology College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China"},{"name":"The Cooperative Innovation Center for Sustainable Pig Production, Huazhong Agricultural University, Wuhan 430000, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qiao","family":"Hu","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Agricultural Microbiology College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China"},{"name":"The Cooperative Innovation Center for Sustainable Pig Production, Huazhong Agricultural University, Wuhan 430000, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Siqi","family":"Chen","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Agricultural Microbiology College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tong","family":"Zhou","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Agricultural Microbiology College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xuexiang","family":"Yu","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Agricultural Microbiology College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China"},{"name":"The Cooperative Innovation Center for Sustainable Pig Production, Huazhong Agricultural University, Wuhan 430000, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hailong","family":"Ma","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Agricultural Microbiology College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China"},{"name":"The Cooperative Innovation Center for Sustainable Pig Production, Huazhong Agricultural University, Wuhan 430000, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ahmed.","family":"H. Ghonaim","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Agricultural Microbiology College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China"},{"name":"The Cooperative Innovation Center for Sustainable Pig Production, Huazhong Agricultural University, Wuhan 430000, China"},{"name":"Desert Research Center, Cairo 11435, Egypt"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hao","family":"Wu","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Agricultural Microbiology College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China"},{"name":"The Cooperative Innovation Center for Sustainable Pig Production, Huazhong Agricultural University, Wuhan 430000, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qi","family":"Sun","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Agricultural Microbiology College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China"},{"name":"The Cooperative Innovation Center for Sustainable Pig Production, Huazhong Agricultural University, Wuhan 430000, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shengxian","family":"Fan","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Agricultural Microbiology College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China"},{"name":"The Cooperative Innovation Center for Sustainable Pig Production, Huazhong Agricultural University, Wuhan 430000, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qigai","family":"He","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Agricultural Microbiology College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China"},{"name":"The Cooperative Innovation Center for Sustainable Pig Production, Huazhong Agricultural University, Wuhan 430000, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2021,4,16]]},"reference":[{"key":"ref_1","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_2","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_3","doi-asserted-by":"crossref","first-page":"79","DOI":"10.1007\/978-1-4939-6869-5_5","article-title":"Generating Recombinant Pseudorabies Virus for Use as a Vaccine Platform","volume":"1581","author":"Tan","year":"2017","journal-title":"Methods Mol. Biol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"285","DOI":"10.1016\/0042-6822(79)90484-7","article-title":"Analysis of the structure of the genome of pseudorabies virus","volume":"95","author":"Rixon","year":"1979","journal-title":"Virology"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"5733","DOI":"10.1016\/j.vaccine.2015.09.066","article-title":"Novel pseudorabies virus variant with defects in TK, gE and gI protects growing pigs against lethal challenge","volume":"33","author":"Hu","year":"2015","journal-title":"Vaccine"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"214","DOI":"10.1186\/s12985-015-0449-3","article-title":"Construction and immune efficacy of recombinant pseudorabies virus expressing PrM-E proteins of Japanese encephalitis virus genotype capital I.; Ukrainian","volume":"12","author":"Qian","year":"2015","journal-title":"Virol. J."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"401","DOI":"10.1007\/s11262-015-1190-0","article-title":"Molecular epidemiology of outbreak-associated pseudorabies virus (PRV) strains in central China","volume":"50","author":"Wang","year":"2015","journal-title":"Virus Genes"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"2432","DOI":"10.1016\/j.vaccine.2015.03.094","article-title":"Construction of a triple gene-deleted Chinese Pseudorabies virus variant and its efficacy study as a vaccine candidate on suckling piglets","volume":"33","author":"Zhang","year":"2015","journal-title":"Vaccine"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"e36","DOI":"10.4142\/jvs.2020.21.e36","article-title":"Epidemiological investigation of porcine pseudorabies virus and its coinfection rate in Shandong Province in China from 2015 to 2018","volume":"21","author":"Ma","year":"2020","journal-title":"J. Vet. Sci."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"550","DOI":"10.1089\/vim.2020.0025","article-title":"Prevalence of Porcine Pseudorabies Virus and Its Coinfection Rate in Heilongjiang Province in China from 2013 to 2018","volume":"33","author":"Zhou","year":"2020","journal-title":"Viral Immunol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"388","DOI":"10.1111\/tbed.13362","article-title":"Dynamics of Aujeszky\u2019s disease virus infection in wild boar in enzootic scenarios","volume":"67","year":"2020","journal-title":"Transbound. Emerg. Dis."},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Pacini, M.I., Forzan, M., Cilia, G., Bernardini, L., Marzoli, F., Pedonese, F., Bandecchi, P., Fratini, F., and Mazzei, M. (2020). Detection of Pseudorabies Virus in Wild Boar Foetus. Animals, 10.","DOI":"10.3390\/ani10020366"},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Liu, Q., Wang, X., Xie, C., Ding, S., Yang, H., Guo, S., Li, J., Qin, L., Ban, F., and Wang, D. (2020). A Novel Human Acute Encephalitis Caused by Pseudorabies Virus Variant Strain. Clin. Infect. Dis.","DOI":"10.1093\/cid\/ciaa987"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"442","DOI":"10.1007\/s13365-019-00822-2","article-title":"Human encephalitis caused by pseudorabies virus infection: A case report","volume":"26","author":"Wang","year":"2020","journal-title":"J. NeuroVirol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"92","DOI":"10.1016\/j.ijid.2019.08.007","article-title":"Characteristics of human encephalitis caused by pseudorabies virus: A case series study","volume":"87","author":"Yang","year":"2019","journal-title":"Int. J. Infect. Dis."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"199","DOI":"10.1038\/79734","article-title":"Dendritic cell regulation of TH1-TH2 development","volume":"1","author":"Moser","year":"2000","journal-title":"Nat. Immunol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"2498","DOI":"10.1038\/s41467-019-10491-8","article-title":"Mesenchymal stem cell therapy induces FLT3L and CD1c+ dendritic cells in systemic lupus erythematosus patients","volume":"10","author":"Yuan","year":"2019","journal-title":"Nat. Commun."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"1982","DOI":"10.4049\/jimmunol.1302391","article-title":"FLT3-Ligand Treatment of Humanized Mice Results in the Generation of Large Numbers of CD141+and CD1c+Dendritic Cells In Vivo","volume":"192","author":"Ding","year":"2014","journal-title":"J. Immunol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"676","DOI":"10.1038\/ni.1615","article-title":"The receptor tyrosine kinase Flt3 is required for dendritic cell development in peripheral lymphoid tissues","volume":"9","author":"Waskow","year":"2008","journal-title":"Nat. Immunol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"1634","DOI":"10.1128\/JVI.01552-10","article-title":"Rabies Virus Expressing Dendritic Cell-Activating Molecules Enhances the Innate and Adaptive Immune Response to Vaccination","volume":"85","author":"Wen","year":"2010","journal-title":"J. Virol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"9642","DOI":"10.1128\/JVI.00326-10","article-title":"Expression of MIP-1\u03b1 (CCL3) by a Recombinant Rabies Virus Enhances Its Immunogenicity by Inducing Innate Immunity and Recruiting Dendritic Cells and B Cells","volume":"84","author":"Zhao","year":"2010","journal-title":"J. Virol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1178","DOI":"10.1038\/s41591-018-0085-8","article-title":"A natural killer\u2013dendritic cell axis defines checkpoint therapy\u2013responsive tumor microenvironments","volume":"24","author":"Barry","year":"2018","journal-title":"Nat. Med."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"3154","DOI":"10.1038\/s41598-018-21575-8","article-title":"CRISPR-Cas9, a tool to efficiently increase the development of recombinant African swine fever viruses","volume":"8","author":"Borca","year":"2018","journal-title":"Sci. Rep."},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Chang, P., Yao, Y., Tang, N., Sadeyen, J.-R., Sealy, J., Clements, A., Bhat, S., Munir, M., Bryant, J.E., and Iqbal, M. (2018). The Application of NHEJ-CRISPR\/Cas9 and Cre-Lox System in the Generation of Bivalent Duck Enteritis Virus Vaccine against Avian Influenza Virus. Viruses, 10.","DOI":"10.3390\/v10020081"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"19176","DOI":"10.1038\/srep19176","article-title":"A CRISPR\/Cas9 and Cre\/Lox system-based express vaccine development strategy against re-emerging Pseudorabies virus","volume":"6","author":"Liang","year":"2016","journal-title":"Sci. Rep."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"816","DOI":"10.1126\/science.1225829","article-title":"A Programmable dual-RNA-guided DNA endonuclease in adaptive bacterial immunity","volume":"337","author":"Jinek","year":"2012","journal-title":"Science"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"823","DOI":"10.1126\/science.1232033","article-title":"RNA-Guided Human Genome Engineering via Cas9","volume":"339","author":"Mali","year":"2013","journal-title":"Science"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"2281","DOI":"10.1038\/nprot.2013.143","article-title":"Genome engineering using the CRISPR-Cas9 system","volume":"8","author":"Ran","year":"2013","journal-title":"Nat. Protoc."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"819","DOI":"10.1126\/science.1231143","article-title":"Multiplex Genome Engineering Using CRISPR\/Cas Systems","volume":"339","author":"Cong","year":"2013","journal-title":"Science"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"467","DOI":"10.1016\/0022-2836(81)90375-2","article-title":"Bacteriophage P1 site-specific recombination. I. Recombination between loxP sites","volume":"150","author":"Sternberg","year":"1981","journal-title":"J. Mol. Biol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"21571","DOI":"10.1021\/acsami.5b06876","article-title":"Antiviral Activity of Graphene Oxide: How Sharp Edged Structure and Charge Matter","volume":"7","author":"Ye","year":"2015","journal-title":"ACS Appl. Mater. Interfaces"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"1195","DOI":"10.1172\/JCI31205","article-title":"DC-based cancer vaccines","volume":"117","author":"Gilboa","year":"2007","journal-title":"J. Clin. Investig."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"77","DOI":"10.1016\/S0022-1759(98)00204-X","article-title":"An advanced culture method for generating large quantities of highly pure dendritic cells from mouse bone marrow","volume":"223","author":"Lutz","year":"1999","journal-title":"J. Immunol. Methods"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"831","DOI":"10.18632\/oncotarget.23160","article-title":"Recombinant rabies virus with the glycoprotein fused with a DC-binding peptide is an efficacious rabies vaccine","volume":"9","author":"Zhang","year":"2017","journal-title":"Oncotarget"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"20","DOI":"10.1016\/j.vetmic.2012.07.002","article-title":"Oral immunization of wild boar and domestic pigs with attenuated live vaccine protects against Pseudorabies virus infection","volume":"161","author":"Maresch","year":"2012","journal-title":"Vet. Microbiol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"287","DOI":"10.1186\/s12917-018-1536-7","article-title":"Safety and immunogenicity of an attenuated Chinese pseudorabies variant by dual deletion of TK&gE genes","volume":"14","author":"Wang","year":"2018","journal-title":"BMC Vet. Res."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"e01614-18","DOI":"10.1128\/JVI.01614-18","article-title":"Virulent Pseudorabies Virus Infection Induces a Specific and Lethal Systemic Inflammatory Response in Mice","volume":"92","author":"Laval","year":"2018","journal-title":"J. Virol."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"3","DOI":"10.1016\/j.vetmic.2016.11.019","article-title":"Vaccines against pseudorabies virus (PrV)","volume":"206","author":"Freuling","year":"2017","journal-title":"Vet. Microbiol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"105876","DOI":"10.1016\/j.intimp.2019.105876","article-title":"The immune-adjuvant activity and the mechanism of resveratrol on pseudorabies virus vaccine in a mouse model","volume":"76","author":"Chen","year":"2019","journal-title":"Int. Immunopharmacol."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"31","DOI":"10.1208\/s12249-018-1245-3","article-title":"Determination of Mycoplasma hyopneumoniae-Specific IgG, IgG1, and IgG2a Titers in BALB\/c Mice Induced by Mineral Oil-Based Oil-in-Water Emulsion Adjuvants Prepared Using a Self-Emulsifying Drug Delivery System","volume":"20","author":"Bastola","year":"2019","journal-title":"AAPS PharmSciTech"},{"key":"ref_41","first-page":"731","article-title":"Aluminium-adjuvanted vaccines\u2014A review of the current state of knowledge","volume":"69","year":"2015","journal-title":"Przeglad Epidemiol."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"102037","DOI":"10.1016\/j.nano.2019.102037","article-title":"Exploring the interactions between engineered nanomaterials and immune cells at 3D nano-bio interfaces to discover potent nano-adjuvants","volume":"21","author":"Ma","year":"2019","journal-title":"Nanomed. Nanotechnol. Biol. Med."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"687","DOI":"10.1016\/j.immuni.2007.11.003","article-title":"How Do Adjuvants Work? Important Considerations for New Generation Adjuvants","volume":"27","author":"McKee","year":"2007","journal-title":"Immunity"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"19","DOI":"10.1186\/s12985-020-1292-8","article-title":"Characterization of a recombinant pseudorabies virus expressing porcine parvovirus VP2 protein and porcine IL-6","volume":"17","author":"Zheng","year":"2020","journal-title":"Virol. J."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"5276","DOI":"10.1016\/j.vaccine.2013.08.052","article-title":"New attenuated vaccine against columnaris disease in fish: Choosing the right parental strain is critical for vaccine efficacy","volume":"31","author":"Mohammed","year":"2013","journal-title":"Vaccine"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"474","DOI":"10.1007\/s11262-016-1324-z","article-title":"Growth characteristics and complete genomic sequence analysis of a novel pseudorabies virus in China","volume":"52","author":"Yu","year":"2016","journal-title":"Virus Genes"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"1705","DOI":"10.1093\/infdis\/jiy731","article-title":"Interspecies Transmission, Genetic Diversity, and Evolutionary Dynamics of Pseudorabies Virus","volume":"219","author":"He","year":"2018","journal-title":"J. Infect. Dis."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"109","DOI":"10.1186\/1743-422X-7-109","article-title":"Attenuation of vaccinia virus by the expression of human Flt3 ligand","volume":"7","author":"Zurkova","year":"2010","journal-title":"Virol. J."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"662","DOI":"10.1007\/s12250-019-00144-x","article-title":"A Recombinant Rabies Virus Expressing Fms-like Tyrosine Kinase 3 Ligand (Flt3L) Induces Enhanced Immunogenicity in Mice","volume":"34","author":"Zhang","year":"2019","journal-title":"Virol. Sin."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"510","DOI":"10.3382\/ps\/pex314","article-title":"Inflammation: Friend or foe for animal production?","volume":"97","author":"Broom","year":"2018","journal-title":"Poult. Sci."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"6299","DOI":"10.1016\/j.vaccine.2008.08.052","article-title":"Vaccine injection technique and reactogenicity\u2014Evidence for practice","volume":"26","year":"2008","journal-title":"Vaccine"}],"container-title":["Viruses"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1999-4915\/13\/4\/691\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T05:48:50Z","timestamp":1760161730000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1999-4915\/13\/4\/691"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,4,16]]},"references-count":51,"journal-issue":{"issue":"4","published-online":{"date-parts":[[2021,4]]}},"alternative-id":["v13040691"],"URL":"https:\/\/doi.org\/10.3390\/v13040691","relation":{},"ISSN":["1999-4915"],"issn-type":[{"value":"1999-4915","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,4,16]]}}}