{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T01:36:30Z","timestamp":1760146590101,"version":"build-2065373602"},"reference-count":52,"publisher":"MDPI AG","issue":"11","license":[{"start":{"date-parts":[[2024,11,19]],"date-time":"2024-11-19T00:00:00Z","timestamp":1731974400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Federal Ministry of Education and Research (BMBF) of the Federal Republic of Germany","award":["01KI2023","GRK2504","TH2166\/1-2"],"award-info":[{"award-number":["01KI2023","GRK2504","TH2166\/1-2"]}]},{"name":"Deutsche Forschungsgemeinschaft (DFG)","award":["01KI2023","GRK2504","TH2166\/1-2"],"award-info":[{"award-number":["01KI2023","GRK2504","TH2166\/1-2"]}]},{"name":"\u201cBavarian Equal Opportunities Sponsorship\u2014Realisierung von Chancengleichheit von Frauen in Forschung und Lehre (FFL)\u2014Realization Equal Opportunities for Women in Research and Teaching\u201d","award":["01KI2023","GRK2504","TH2166\/1-2"],"award-info":[{"award-number":["01KI2023","GRK2504","TH2166\/1-2"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Pathogens"],"abstract":"<jats:p>Human T-cell leukemia virus type 1 (HTLV-1) infects CD4+ T-cells through close cell\u2013cell contacts. The viral Tax-1 (Tax) protein regulates transcription by transactivating the HTLV-1 U3R promoter in the 5\u2032 long terminal repeat of the integrated provirus. Here, we generated a clonal Tax-responsive T-cell line to track HTLV-1 infection at the single-cell level using flow cytometry, bypassing intracellular viral protein staining. Jurkat T-cells stably transduced with the SMPU vector carrying green fluorescent protein (GFP) under control of 18 \u00d7 21 bp Tax-responsive element repeats of the U3R were evaluated. Among 40 clones analyzed for Tax responsiveness, the top two were characterized. Upon overexpression of Tax, over 40% of the cells showed GFP positivity, and approximately 90% of the Tax-positive cells were GFP-positive, indicating efficient reporter activity. However, with CREB-deficient Tax mutant M47, both total GFP-positive cell counts and those within the Tax-positive group significantly decreased. Co-culture with chronically HTLV-1-infected MT-2 or C91-PL cells led to an average of 0.9% or 2.4% GFP-positive cells, respectively, confirming the suitability to monitor HTLV-1 transmission and that HTLV-1 infection is very low. Thus, the novel Tax-responsive reporter T-cell line is a suitable tool to monitor infection of HTLV-1 on the single-cell level.<\/jats:p>","DOI":"10.3390\/pathogens13111015","type":"journal-article","created":{"date-parts":[[2024,11,19]],"date-time":"2024-11-19T04:53:05Z","timestamp":1731991985000},"page":"1015","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["A Novel Tax-Responsive Reporter T-Cell Line to Analyze Infection of HTLV-1"],"prefix":"10.3390","volume":"13","author":[{"given":"Stefanie","family":"Heym","sequence":"first","affiliation":[{"name":"Institute of Clinical and Molecular Virology, Friedrich-Alexander-Universit\u00e4t Erlangen-N\u00fcrnberg (FAU), 91054 Erlangen, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Pauline","family":"Krebs","sequence":"additional","affiliation":[{"name":"Institute of Clinical and Molecular Virology, Friedrich-Alexander-Universit\u00e4t Erlangen-N\u00fcrnberg (FAU), 91054 Erlangen, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kristin","family":"Ott","sequence":"additional","affiliation":[{"name":"Institute of Clinical and Molecular Virology, Friedrich-Alexander-Universit\u00e4t Erlangen-N\u00fcrnberg (FAU), 91054 Erlangen, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Norbert","family":"Donhauser","sequence":"additional","affiliation":[{"name":"Institute of Clinical and Molecular Virology, Friedrich-Alexander-Universit\u00e4t Erlangen-N\u00fcrnberg (FAU), 91054 Erlangen, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Laura M.","family":"Kemeter","sequence":"additional","affiliation":[{"name":"Institute of Clinical and Molecular Virology, Friedrich-Alexander-Universit\u00e4t Erlangen-N\u00fcrnberg (FAU), 91054 Erlangen, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2225-0081","authenticated-orcid":false,"given":"Florian","family":"Simon","sequence":"additional","affiliation":[{"name":"Institute of Clinical and Molecular Virology, Friedrich-Alexander-Universit\u00e4t Erlangen-N\u00fcrnberg (FAU), 91054 Erlangen, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sebastian","family":"Millen","sequence":"additional","affiliation":[{"name":"Institute of Clinical and Molecular Virology, Friedrich-Alexander-Universit\u00e4t Erlangen-N\u00fcrnberg (FAU), 91054 Erlangen, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8568-9526","authenticated-orcid":false,"given":"Andrea K.","family":"Thoma-Kress","sequence":"additional","affiliation":[{"name":"Institute of Clinical and Molecular Virology, Friedrich-Alexander-Universit\u00e4t Erlangen-N\u00fcrnberg (FAU), 91054 Erlangen, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2024,11,19]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1893","DOI":"10.1016\/S0140-6736(18)30974-7","article-title":"Time to eradicate HTLV-1: An open letter to WHO","volume":"391","author":"Martin","year":"2018","journal-title":"Lancet"},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Gross, C., and Thoma-Kress, A.K. (2016). Molecular Mechanisms of HTLV-1 Cell-to-Cell Transmission. Viruses, 8.","DOI":"10.3390\/v8030074"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1713","DOI":"10.1126\/science.1080115","article-title":"Spread of HTLV-I between lymphocytes by virus-induced polarization of the cytoskeleton","volume":"299","author":"Igakura","year":"2003","journal-title":"Science"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"83","DOI":"10.1038\/nm.2065","article-title":"Biofilm-like extracellular viral assemblies mediate HTLV-1 cell-to-cell transmission at virological synapses","volume":"16","author":"Sachse","year":"2010","journal-title":"Nat. Med."},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Donhauser, N., Socher, E., Millen, S., Heym, S., Sticht, H., and Thoma-Kress, A.K. (2020). Transfer of HTLV-1 p8 and Gag to target T-cells depends on VASP, a novel interaction partner of p8. PLoS Pathog., 16.","DOI":"10.1371\/journal.ppat.1008879"},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Omsland, M., Pise-Masison, C., Fujikawa, D., Galli, V., Fenizia, C., Parks, R.W., Gjertsen, B.T., Franchini, G., and Andresen, V. (2018). Inhibition of Tunneling Nanotube (TNT) Formation and Human T-cell Leukemia Virus Type 1 (HTLV-1) Transmission by Cytarabine. Sci. Rep., 8.","DOI":"10.1038\/s41598-018-29391-w"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"20738","DOI":"10.1073\/pnas.1009635107","article-title":"Human T-cell leukemia virus type 1 p8 protein increases cellular conduits and virus transmission","volume":"107","author":"Gold","year":"2010","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"9922","DOI":"10.1128\/JVI.77.18.9922-9930.2003","article-title":"Human T-cell leukemia virus type 1 envelope glycoprotein gp46 interacts with cell surface heparan sulfate proteoglycans","volume":"77","author":"Pinon","year":"2003","journal-title":"J. Virol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"449","DOI":"10.1016\/S0092-8674(03)00881-X","article-title":"The ubiquitous glucose transporter GLUT-1 is a receptor for HTLV","volume":"115","author":"Manel","year":"2003","journal-title":"Cell"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"6844","DOI":"10.1128\/JVI.02719-05","article-title":"Neuropilin-1 is involved in human T-cell lymphotropic virus type 1 entry","volume":"80","author":"Ghez","year":"2006","journal-title":"J. Virol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"31","DOI":"10.1186\/1742-4690-4-31","article-title":"Isolated receptor binding domains of HTLV-1 and HTLV-2 envelopes bind Glut-1 on activated CD4+ and CD8+ T cells","volume":"4","author":"Kinet","year":"2007","journal-title":"Retrovirology"},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Currer, R., Van Duyne, R., Jaworski, E., Guendel, I., Sampey, G., Das, R., Narayanan, A., and Kashanchi, F. (2012). HTLV tax: A fascinating multifunctional co-regulator of viral and cellular pathways. Front. Microbiol., 3.","DOI":"10.3389\/fmicb.2012.00406"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"2175","DOI":"10.1128\/jvi.61.7.2175-2181.1987","article-title":"Identification of p40x-responsive regulatory sequences within the human T-cell leukemia virus type I long terminal repeat","volume":"61","author":"Brady","year":"1987","journal-title":"J. Virol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"6502","DOI":"10.1073\/pnas.82.19.6502","article-title":"Location of cis-acting regulatory sequences in the human T-cell leukemia virus type I long terminal repeat","volume":"82","author":"Rosen","year":"1985","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"5207","DOI":"10.1093\/nar\/17.13.5207","article-title":"Differential activation of the 21-base-pair enhancer element of human T-cell leukemia virus type I by its own trans-activator and cyclic AMP","volume":"17","author":"Nakamura","year":"1989","journal-title":"Nucleic Acids Res."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1604","DOI":"10.1128\/jvi.63.4.1604-1611.1989","article-title":"Response of the human T-cell leukemia virus type 1 long terminal repeat to cyclic AMP","volume":"63","author":"Poteat","year":"1989","journal-title":"J. Virol."},{"key":"ref_17","first-page":"361","article-title":"Tax-independent binding of multiple cellular factors to Tax-response element DNA of HTLV-I","volume":"5","author":"Beimling","year":"1990","journal-title":"Oncogene"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"2537","DOI":"10.1002\/j.1460-2075.1990.tb07434.x","article-title":"Multiple cDNA clones encoding nuclear proteins that bind to the tax-dependent enhancer of HTLV-1: All contain a leucine zipper structure and basic amino acid domain","volume":"9","author":"Yoshimura","year":"1990","journal-title":"EMBO J."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"7070","DOI":"10.1073\/pnas.89.15.7070","article-title":"Human T-cell lymphotropic virus type I (HTLV-I) transcriptional activator, Tax, enhances CREB binding to HTLV-I 21-base-pair repeats by protein-protein interaction","volume":"89","author":"Zhao","year":"1992","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"610","DOI":"10.1073\/pnas.90.2.610","article-title":"The trans-activator tax of human T-cell leukemia virus type 1 (HTLV-1) interacts with cAMP-responsive element (CRE) binding and CRE modulator proteins that bind to the 21-base-pair enhancer of HTLV-1","volume":"90","author":"Suzuki","year":"1993","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"642","DOI":"10.1038\/380642a0","article-title":"Control of cAMP-regulated enhancers by the viral transactivator Tax through CREB and the co-activator CBP","volume":"380","author":"Kwok","year":"1996","journal-title":"Nature"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1016","DOI":"10.1182\/blood-2008-03-136770","article-title":"HTLV-1-Tax and ICAM-1 act on T-cell signal pathways to polarize the microtubule-organizing center at the virological synapse","volume":"114","author":"Nejmeddine","year":"2009","journal-title":"Blood"},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Gross, C., Wiesmann, V., Millen, S., Kalmer, M., Wittenberg, T., Gettemans, J., and Thoma-Kress, A.K. (2016). The Tax-Inducible Actin-Bundling Protein Fascin Is Crucial for Release and Cell-to-Cell Transmission of Human T-Cell Leukemia Virus Type 1 (HTLV-1). PLoS Pathog., 12.","DOI":"10.1371\/journal.ppat.1005916"},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Arone, C., Martial, S., Burlaud-Gaillard, J., Thoulouze, M.I., Roingeard, P., Dutartre, H., and Muriaux, D. (2023). HTLV-1 biofilm polarization maintained by tetraspanin CD82 is required for efficient viral transmission. mBio, 14.","DOI":"10.1128\/mbio.01326-23"},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Chevalier, S.A., Turpin, J., Cachat, A., Afonso, P.V., Gessain, A., Brady, J.N., Pise-Masison, C.A., and Mahieux, R. (2014). Gem-induced cytoskeleton remodeling increases cellular migration of HTLV-1-infected cells, formation of infected-to-target T-cell conjugates and viral transmission. PLoS Pathog., 10.","DOI":"10.1371\/journal.ppat.1003917"},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Hiyoshi, M., Takahashi, N., Eltalkhawy, Y.M., Noyori, O., Lotfi, S., Panaampon, J., Okada, S., Tanaka, Y., Ueno, T., and Fujisawa, J.I. (2021). M-Sec induced by HTLV-1 mediates an efficient viral transmission. PLoS Pathog., 17.","DOI":"10.1371\/journal.ppat.1010126"},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"Millen, S., Gross, C., Donhauser, N., Mann, M.C., Peloponese, J.M., and Thoma-Kress, A.K. (2019). Collagen IV (COL4A1, COL4A2), a Component of the Viral Biofilm, Is Induced by the HTLV-1 Oncoprotein Tax and Impacts Virus Transmission. Front. Microbiol., 10.","DOI":"10.3389\/fmicb.2019.02439"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"10580","DOI":"10.1128\/JVI.01799-15","article-title":"Viral Source-Independent High Susceptibility of Dendritic Cells to Human T-Cell Leukemia Virus Type 1 Infection Compared to That of T Lymphocytes","volume":"89","author":"Alais","year":"2015","journal-title":"J. Virol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"10","DOI":"10.1128\/JVI.00608-19","article-title":"Human T-Cell Leukemia Virus Type 1 (HTLV-1) bZIP Factor Upregulates the Expression of ICAM-1 To Facilitate HTLV-1 Infection","volume":"93","author":"Fazio","year":"2019","journal-title":"J. Virol."},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"Hatayama, Y., Yamaoka, Y., Morita, T., Jeremiah, S.S., Miyakawa, K., Nishi, M., Kimura, Y., Mitsunaga, M., Iwase, T., and Kimura, H. (2022). Development of a Monoclonal Antibody Targeting HTLV-1 Envelope gp46 Glycoprotein and Its Application to Near-Infrared Photoimmuno-Antimicrobial Strategy. Viruses, 14.","DOI":"10.3390\/v14102153"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"7459","DOI":"10.1128\/JVI.00130-06","article-title":"Versatile reporter systems show that transactivation by human T-cell leukemia virus type 1 Tax occurs independently of chromatin remodeling factor BRG1","volume":"80","author":"Zhang","year":"2006","journal-title":"J. Virol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"5402","DOI":"10.1073\/pnas.80.17.5402","article-title":"Transformation of human umbilical cord blood T cells by human T-cell leukemia\/lymphoma virus","volume":"80","author":"Popovic","year":"1983","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"4576","DOI":"10.1182\/blood-2002-09-2986","article-title":"Arsenic\/interferon specifically reverses 2 distinct gene networks critical for the survival of HTLV-1-infected leukemic cells","volume":"101","author":"Nasr","year":"2003","journal-title":"Blood"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"1875","DOI":"10.1101\/gad.4.11.1875","article-title":"Identification of HTLV-I tax trans-activator mutants exhibiting novel transcriptional phenotypes","volume":"4","author":"Smith","year":"1990","journal-title":"Genes Dev."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"30","DOI":"10.1186\/s12977-020-00538-w","article-title":"A novel positive feedback-loop between the HTLV-1 oncoprotein Tax and NF-kappaB activity in T-cells","volume":"17","author":"Millen","year":"2020","journal-title":"Retrovirology"},{"key":"ref_36","first-page":"295","article-title":"Development and characterization of monoclonal antibodies to the HTLV-I Tax (P40X) protein","volume":"8","author":"Langton","year":"1988","journal-title":"Med. Virol."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"2266","DOI":"10.1016\/j.ymthe.2023.03.014","article-title":"Precise excision of HTLV-1 provirus with a designer-recombinase","volume":"31","author":"Karpinski","year":"2023","journal-title":"Mol. Ther."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"29","DOI":"10.1016\/j.ymeth.2015.02.016","article-title":"Appraising the suitability of succinimidyl and lipophilic fluorescent dyes to track proliferation in non-quiescent cells by dye dilution","volume":"82","author":"Filby","year":"2015","journal-title":"Methods"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"101","DOI":"10.1007\/978-1-0716-2376-3_9","article-title":"CellTrace Violet Flow Cytometric Assay to Assess Cell Proliferation","volume":"2508","author":"Lemieszek","year":"2022","journal-title":"Methods Mol. Biol."},{"key":"ref_40","doi-asserted-by":"crossref","unstructured":"Mazurov, D., Ilinskaya, A., Heidecker, G., Lloyd, P., and Derse, D. (2010). Quantitative comparison of HTLV-1 and HIV-1 cell-to-cell infection with new replication dependent vectors. PLoS Pathog., 6.","DOI":"10.1371\/journal.ppat.1000788"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"e2005568118","DOI":"10.1073\/pnas.2005568118","article-title":"NF-kappaB-induced R-loop accumulation and DNA damage select for nucleotide excision repair deficiencies in adult T cell leukemia","volume":"118","author":"He","year":"2021","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"11106","DOI":"10.1128\/JVI.75.22.11106-11115.2001","article-title":"Human T-cell leukemia virus type 1 Tax protein binds to assembled nuclear proteasomes and enhances their proteolytic activity","volume":"75","author":"Hemelaar","year":"2001","journal-title":"J. Virol."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"316","DOI":"10.1016\/0042-6822(92)90320-O","article-title":"Characterization of a novel nuclear localization signal in the HTLV-I tax transactivator protein","volume":"187","author":"Smith","year":"1992","journal-title":"Virology"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"1035","DOI":"10.1093\/protein\/12.12.1035","article-title":"Attenuation of green fluorescent protein half-life in mammalian cells","volume":"12","author":"Corish","year":"1999","journal-title":"Protein Eng."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"720","DOI":"10.1073\/pnas.0507631103","article-title":"HTLV-I basic leucine zipper factor gene mRNA supports proliferation of adult T cell leukemia cells","volume":"103","author":"Satou","year":"2006","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"1283","DOI":"10.4049\/jimmunol.143.4.1283","article-title":"Mechanisms of T cell activation by the calcium ionophore ionomycin","volume":"143","author":"Chatila","year":"1989","journal-title":"J. Immunol."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"821","DOI":"10.1146\/annurev.biochem.68.1.821","article-title":"CREB: A stimulus-induced transcription factor activated by a diverse array of extracellular signals","volume":"68","author":"Shaywitz","year":"1999","journal-title":"Annu. Rev. Biochem."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"6","DOI":"10.1186\/s12977-021-00550-8","article-title":"Extracellular vesicles from HTLV-1 infected cells modulate target cells and viral spread","volume":"18","author":"Pinto","year":"2021","journal-title":"Retrovirology"},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"13386","DOI":"10.1128\/JVI.02758-13","article-title":"Alpha interferon restricts human T-lymphotropic virus type 1 and 2 de novo infection through PKR activation","volume":"87","author":"Cachat","year":"2013","journal-title":"J. Virol."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"10591","DOI":"10.1128\/JVI.01940-15","article-title":"Improvement of HIV-1 and Human T Cell Lymphotropic Virus Type 1 Replication-Dependent Vectors via Optimization of Reporter Gene Reconstitution and Modification with Intronic Short Hairpin RNA","volume":"89","author":"Shunaeva","year":"2015","journal-title":"J. Virol."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"8461","DOI":"10.1128\/JVI.75.18.8461-8468.2001","article-title":"Examining human T-lymphotropic virus type 1 infection and replication by cell-free infection with recombinant virus vectors","volume":"75","author":"Derse","year":"2001","journal-title":"J. Virol."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"26","DOI":"10.1186\/1742-4690-10-26","article-title":"Restriction of diverse retroviruses by SAMHD1","volume":"10","author":"Gramberg","year":"2013","journal-title":"Retrovirology"}],"container-title":["Pathogens"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-0817\/13\/11\/1015\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T16:35:03Z","timestamp":1760114103000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-0817\/13\/11\/1015"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,11,19]]},"references-count":52,"journal-issue":{"issue":"11","published-online":{"date-parts":[[2024,11]]}},"alternative-id":["pathogens13111015"],"URL":"https:\/\/doi.org\/10.3390\/pathogens13111015","relation":{},"ISSN":["2076-0817"],"issn-type":[{"type":"electronic","value":"2076-0817"}],"subject":[],"published":{"date-parts":[[2024,11,19]]}}}