{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,5]],"date-time":"2026-05-05T01:38:59Z","timestamp":1777945139998,"version":"3.51.4"},"reference-count":66,"publisher":"Wiley","issue":"3","license":[{"start":{"date-parts":[[2026,4,23]],"date-time":"2026-04-23T00:00:00Z","timestamp":1776902400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"},{"start":{"date-parts":[[2026,4,23]],"date-time":"2026-04-23T00:00:00Z","timestamp":1776902400000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["J of Molecular Recognition"],"published-print":{"date-parts":[[2026,5]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>\n                    Canine parvovirus type 2 (CPV\u20102) is a highly contagious and often fatal canine pathogen, with antigenic variants (CPV\u20102a, 2b, 2c), maternally derived antibody (MDA) interference, and lack of Differentiating Infected from Vaccinated Animals (DIVA) capability limiting current vaccines. Using an immunoinformatics\u2010based reverse vaccinology approach, we identified conserved B\u2010cell, cytotoxic T lymphocyte (CTL), and helper T lymphocyte (HTL) epitopes from CPV\u20102 structural (VP1, VP2) and nonstructural (NS1, NS2) proteins. These were screened for antigenicity, immunogenicity, safety, and stability. The screening identified three B\u2010cell, two CTL, and five HTL epitopes. Out of these, two representative epitopes were utilized for in\u00a0vitro evaluation. The peripheral blood mononuclear cells (PBMCs) were collected from 10 dogs 28\u2009days after vaccination with a live\u2010attenuated whole\u2010virus CPV vaccine and pooled. This pool was expected to contain memory cytotoxic T cells recognizing a broad viral epitope repertoire. On antigenic stimulation, PCPV2 induced significant proliferation of canine PBMCs, with the highest response at 33.33\u2009\u03bcg\/mL, alongside ~15.67\u2010fold upregulation of IFN\u2010\u03b3 mRNA, indicating a memory T\u2010cell response. A multi\u2010epitope vaccine was designed by integrating validated epitopes with the TLR\u20104 agonist RS09 and a Capripoxvirus\u2010derived DIVA marker, linked by AAY\/EAAAK spacers. The vaccine construct can be effectively delivered via a recombinant canine adenovirus\u20102 vector for efficient in\u00a0vivo expression and cellular response. Structural modeling, molecular docking, immune simulations, molecular dynamic simulations, and codon optimization predicted strong receptor binding, stability, IgG\/T\u2010cell responses, and expression feasibility in\n                    <jats:styled-content style=\"fixed-case\">\n                      <jats:italic>Escherichia coli<\/jats:italic>\n                    <\/jats:styled-content>\n                    and canine cells. These findings support PCPV2 as a potent immunogen and demonstrate the promise of the designed construct for a potential adenovirus\u2010vectored vaccine; however, in\u00a0vivo validation will be essential to confirm efficacy, cross\u2010variant protection, and DIVA performance.\n                  <\/jats:p>","DOI":"10.1002\/jmr.70036","type":"journal-article","created":{"date-parts":[[2026,4,24]],"date-time":"2026-04-24T02:05:13Z","timestamp":1776996313000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Design of Multi\u2010Epitope\n                    <scp>DIVA<\/scp>\n                    \u2010Compatible Vaccine Candidate Against Canine Parvovirus 2"],"prefix":"10.1002","volume":"39","author":[{"given":"Shubham","family":"Sharma","sequence":"first","affiliation":[{"name":"Department of Veterinary Pharmacology RAJUVAS  Bikaner Rajasthan India"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Pratishtha","family":"Sharma","sequence":"additional","affiliation":[{"name":"Department of Veterinary Pharmacology RAJUVAS  Bikaner Rajasthan India"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ravindra Pal","family":"Mandar","sequence":"additional","affiliation":[{"name":"Department of Veterinary Pharmacology RAJUVAS  Bikaner Rajasthan India"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ram","family":"Kumar","sequence":"additional","affiliation":[{"name":"Department of Veterinary Microbiology RAJUVAS  Bikaner Rajasthan India"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ashok","family":"Gaur","sequence":"additional","affiliation":[{"name":"Department of Veterinary Pharmacology RAJUVAS  Bikaner Rajasthan India"},{"name":"Center for Ethno\u2010Veterinary Practice and Alternative Medicines (CEVPAM) RAJUVAS  Bikaner Rajasthan India"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mamta","family":"Gurjar","sequence":"additional","affiliation":[{"name":"Department of Veterinary Microbiology RAJUVAS  Bikaner Rajasthan India"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Rajesh","family":"Singathia","sequence":"additional","affiliation":[{"name":"Department of Veterinary Microbiology RAJUVAS  Bikaner Rajasthan India"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Braj Nandan","family":"Shringi","sequence":"additional","affiliation":[{"name":"Department of Veterinary Microbiology RAJUVAS  Bikaner Rajasthan India"},{"name":"Vaccinology and Biological Products Center (VBPRC) RAJUVAS  Bikaner Rajasthan India"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sunita","family":"Choudhary","sequence":"additional","affiliation":[{"name":"Department of Veterinary Medicine RAJUVAS  Bikaner Rajasthan India"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2139-4542","authenticated-orcid":false,"given":"Amit Kumar","family":"Pandey","sequence":"additional","affiliation":[{"name":"Vaccinology and Biological Products Center (VBPRC) RAJUVAS  Bikaner Rajasthan India"},{"name":"Department of Veterinary Biochemistry RAJUVAS  Bikaner Rajasthan India"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2026,4,23]]},"reference":[{"key":"e_1_2_10_2_1","doi-asserted-by":"publisher","DOI":"10.1007\/s13337\u2010010\u20100007\u2010y"},{"key":"e_1_2_10_3_1","doi-asserted-by":"publisher","DOI":"10.18805\/IJAR.B\u20104302"},{"key":"e_1_2_10_4_1","doi-asserted-by":"publisher","DOI":"10.1101\/cshperspect.a012989"},{"key":"e_1_2_10_5_1","doi-asserted-by":"publisher","DOI":"10.1051\/vetres\/2010011"},{"key":"e_1_2_10_6_1","doi-asserted-by":"publisher","DOI":"10.1111\/jsap.13718"},{"key":"e_1_2_10_7_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.anireprosci.2019.06.012"},{"key":"e_1_2_10_8_1","doi-asserted-by":"publisher","DOI":"10.3390\/ani11113260"},{"key":"e_1_2_10_9_1","doi-asserted-by":"publisher","DOI":"10.3390\/vaccines11091499"},{"key":"e_1_2_10_10_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.vetmic.2020.108760"},{"key":"e_1_2_10_11_1","doi-asserted-by":"publisher","DOI":"10.3390\/biomedicines11082180"},{"key":"e_1_2_10_12_1","doi-asserted-by":"publisher","DOI":"10.1007\/s42770\u2010024\u201001442\u20107"},{"key":"e_1_2_10_13_1","doi-asserted-by":"publisher","DOI":"10.1007\/978\u20101\u20100716\u20101884\u20104_6"},{"key":"e_1_2_10_14_1","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkab1112"},{"key":"e_1_2_10_15_1","doi-asserted-by":"publisher","DOI":"10.36106\/ijsr"},{"key":"e_1_2_10_16_1","doi-asserted-by":"publisher","DOI":"10.14202\/vetworld.2025.2273\u20102286"},{"key":"e_1_2_10_17_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jbi.2019.103160"},{"key":"e_1_2_10_18_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.vaccine.2025.127075"},{"key":"e_1_2_10_19_1","doi-asserted-by":"publisher","DOI":"10.1093\/bioinformatics\/17.7.646"},{"key":"e_1_2_10_20_1","doi-asserted-by":"publisher","DOI":"10.1006\/jmbi.2000.4315"},{"key":"e_1_2_10_21_1","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkx346"},{"key":"e_1_2_10_22_1","doi-asserted-by":"publisher","DOI":"10.1186\/1745\u20107580\u20106\u20106"},{"key":"e_1_2_10_23_1","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gku938"},{"key":"e_1_2_10_24_1","doi-asserted-by":"publisher","DOI":"10.1093\/bioinformatics\/btv639"},{"key":"e_1_2_10_25_1","doi-asserted-by":"publisher","DOI":"10.1186\/1471\u20102105\u20108\u20104"},{"key":"e_1_2_10_26_1","doi-asserted-by":"publisher","DOI":"10.1093\/bioinformatics\/btt619"},{"key":"e_1_2_10_27_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0073957"},{"key":"e_1_2_10_28_1","doi-asserted-by":"publisher","DOI":"10.1007\/s10822\u2010016\u20109902\u20107"},{"key":"e_1_2_10_29_1","doi-asserted-by":"publisher","DOI":"10.1385\/1-59259-890-0:571"},{"key":"e_1_2_10_30_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.compbiomed.2023.106929"},{"key":"e_1_2_10_31_1","doi-asserted-by":"publisher","DOI":"10.1186\/1745\u20106150\u20108\u201030"},{"key":"e_1_2_10_32_1","doi-asserted-by":"publisher","DOI":"10.1038\/srep42851"},{"key":"e_1_2_10_33_1","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2022.998262"},{"key":"e_1_2_10_34_1","doi-asserted-by":"publisher","DOI":"10.33011\/livecoms.1.1.5068"},{"key":"e_1_2_10_35_1","doi-asserted-by":"publisher","DOI":"10.3329\/bjvm.v14i2.31403"},{"issue":"2","key":"e_1_2_10_36_1","first-page":"83","article-title":"Epidemiological Study of Parvo Gastroenteritis and Its Management by Different Concentrations of Fluid in Pups","volume":"4","author":"Haque S.","year":"2012","journal-title":"Indian Journal of Canine Practice"},{"key":"e_1_2_10_37_1","doi-asserted-by":"publisher","DOI":"10.1155\/2021\/8862180"},{"key":"e_1_2_10_38_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jgeb.2025.100510"},{"key":"e_1_2_10_39_1","doi-asserted-by":"publisher","DOI":"10.1186\/s12917\u2010024\u201004006\u2010x"},{"key":"e_1_2_10_40_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0244176"},{"key":"e_1_2_10_41_1","doi-asserted-by":"publisher","DOI":"10.1007\/978\u20100\u2010387\u201079208\u20108_3"},{"key":"e_1_2_10_42_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0022011"},{"key":"e_1_2_10_43_1","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.1502018"},{"key":"e_1_2_10_44_1","doi-asserted-by":"publisher","DOI":"10.3389\/fcimb.2023.1309096"},{"key":"e_1_2_10_45_1","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2023.1280299"},{"key":"e_1_2_10_46_1","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.180.9.5771"},{"key":"e_1_2_10_47_1","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2022.872155"},{"key":"e_1_2_10_48_1","first-page":"49","article-title":"Cloning and Sequencing of Bovine Interleukin\u20101\u03b2","volume":"14","author":"Pandey A.","year":"2013","journal-title":"Veterinary Practioner"},{"key":"e_1_2_10_49_1","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2021.712488"},{"key":"e_1_2_10_50_1","doi-asserted-by":"publisher","DOI":"10.1021\/acs.molpharmaceut.5b00802"},{"key":"e_1_2_10_51_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0285177"},{"key":"e_1_2_10_52_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0030839"},{"key":"e_1_2_10_53_1","doi-asserted-by":"publisher","DOI":"10.1155\/vmi\/9191117"},{"key":"e_1_2_10_54_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jmgm.2020.107645"},{"key":"e_1_2_10_55_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0335147"},{"key":"e_1_2_10_56_1","doi-asserted-by":"crossref","unstructured":"Y.Zhu X.Liu Y.Xu G.Zhang Y.Yin andC.Wu \u201cConstruction and Immunogenicity Detection of Canine Parvovirus\u2010Like Particles Fusion With Canine Febrile Antigen Epitopes \u201d Preprint bioRxiv January 2026 https:\/\/doi.org\/10.64898\/2026.01.18.700204.","DOI":"10.64898\/2026.01.18.700204"},{"key":"e_1_2_10_57_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.vetmic.2026.110933"},{"key":"e_1_2_10_58_1","doi-asserted-by":"publisher","DOI":"10.3389\/fvets.2025.1624275"},{"key":"e_1_2_10_59_1","doi-asserted-by":"publisher","DOI":"10.4161\/onci.21355"},{"key":"e_1_2_10_60_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1348\u20100421.2008.00032.x"},{"key":"e_1_2_10_61_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0294663"},{"key":"e_1_2_10_62_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41598\u2010021\u201003932\u20102"},{"key":"e_1_2_10_63_1","doi-asserted-by":"publisher","DOI":"10.4103\/jgid.jgid_96_21"},{"key":"e_1_2_10_64_1","doi-asserted-by":"publisher","DOI":"10.24171\/j.phrp.2024.0013"},{"key":"e_1_2_10_65_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41598\u2010025\u201016143\u2010w"},{"key":"e_1_2_10_66_1","doi-asserted-by":"publisher","DOI":"10.1002\/jmr.3007"},{"key":"e_1_2_10_67_1","doi-asserted-by":"publisher","DOI":"10.1007\/s11259\u2010025\u201010877\u20108"}],"container-title":["Journal of Molecular Recognition"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/jmr.70036","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/full-xml\/10.1002\/jmr.70036","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/jmr.70036","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,5,2]],"date-time":"2026-05-02T09:38:46Z","timestamp":1777714726000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1002\/jmr.70036"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,4,23]]},"references-count":66,"journal-issue":{"issue":"3","published-print":{"date-parts":[[2026,5]]}},"alternative-id":["10.1002\/jmr.70036"],"URL":"https:\/\/doi.org\/10.1002\/jmr.70036","archive":["Portico"],"relation":{},"ISSN":["0952-3499","1099-1352"],"issn-type":[{"value":"0952-3499","type":"print"},{"value":"1099-1352","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,4,23]]},"assertion":[{"value":"2025-09-07","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-04-10","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-04-23","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"e70036"}}