{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,9]],"date-time":"2026-09-09T21:01:34Z","timestamp":1788987694305,"version":"build-2803163510"},"reference-count":47,"publisher":"MDPI AG","issue":"9","license":[{"start":{"date-parts":[[2025,9,22]],"date-time":"2025-09-22T00:00:00Z","timestamp":1758499200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"\u201c14th Five-Year Plan\u201d National Key R&amp;D Program of China","award":["2023YD1802"],"award-info":[{"award-number":["2023YD1802"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Veterinary Sciences"],"abstract":"<jats:p>The widespread prevalence of different serotypes of fowl adenoviruses (FAdVs) has led to diverse vaccine demands, especially for subunit vaccines targeting FAdV-4 and FAdV-11, which cause hydropericardium-hepatitis syndrome (HHS) and inclusion body hepatitis (IBH), respectively. Although the Fiber protein is known to elicit robust immune protection, further exploration is needed to enhance the production of cross-neutralizing antibodies. This study utilized structural prediction and homology modeling techniques, employing domain-swapping strategy to integrate neutralizing epitope-containing amino acid sequences (274\u2013451aa and 364\u2013543aa) into the shaft domain of the Fiber protein. Two novel chimeric proteins were recombinantly expressed and developed into subunit vaccines: Fiber-C4\/D11 and Fiber-D11\/C4. Immunogenicity assessments revealed that the Fiber-C4\/D11 vaccine group rapidly induced an antibody response against FAdV-11 within 7 days post-vaccination. By 28 days post-vaccination (dpv), the Fiber-C4\/D11 vaccine group exhibited significantly higher levels of cross-neutralizing antibodies compared to the Fiber-D11\/C4 group (p &lt; 0.05). Challenge experiments demonstrated that both vaccines effectively alleviated clinical symptoms and prevented mortality in SPF chickens. Compared to Fiber-D11\/C4, Fiber-C4\/D11 significantly reduced body weight loss, liver lesions, viral titers in tissues, and viral shedding. Notably, no cross-neutralizing antibodies were detected following FAdV-4 or FAdV-11 infection, indicating a lack of natural cross-protection between the two serotypes. The chimeric vaccine addressed this gap, offering a promising multivalent approach to mitigate FAdV infections and advancing fowl adenoviral subunit vaccine strategies.<\/jats:p>","DOI":"10.3390\/vetsci12090920","type":"journal-article","created":{"date-parts":[[2025,9,22]],"date-time":"2025-09-22T17:08:17Z","timestamp":1758560897000},"page":"920","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["A Novel Chimeric Fiber-C4\/D11 Subunit Vaccine Induces Cross-Neutralizing Antibodies and Provides Better Protection Against Fowl Adenovirus (FAdV) Type 4 and Type 11 Than the Fiber-D11\/C4 Subunit Vaccine"],"prefix":"10.3390","volume":"12","author":[{"given":"Xiangqin","family":"Wang","sequence":"first","affiliation":[{"name":"College of Veterinary Medicine, Hebei Agricultural University, Baoding 071001, China"},{"name":"National Research Center of Engineering and Technology for Veterinary Biologicals, Nanjing 210014, China"},{"name":"Group Biological Products R and D Center, Shandong Sinder Technology Co., Ltd., Qingdao 266100, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kuan","family":"Zhao","sequence":"additional","affiliation":[{"name":"College of Veterinary Medicine, Hebei Agricultural University, Baoding 071001, China"},{"name":"National Research Center of Engineering and Technology for Veterinary Biologicals, Nanjing 210014, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Baishi","family":"Lei","sequence":"additional","affiliation":[{"name":"College of Veterinary Medicine, Hebei Agricultural University, Baoding 071001, China"},{"name":"National Research Center of Engineering and Technology for Veterinary Biologicals, Nanjing 210014, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1946-7212","authenticated-orcid":false,"given":"Wenming","family":"Jiang","sequence":"additional","affiliation":[{"name":"Laboratory of Surveillance for Avian Diseases, China Animal Health and Epidemiology Center, No. 369 Nanjing Road, Qingdao 266032, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yanliang","family":"Qiao","sequence":"additional","affiliation":[{"name":"Group Biological Products R and D Center, Shandong Sinder Technology Co., Ltd., Qingdao 266100, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wanzhe","family":"Yuan","sequence":"additional","affiliation":[{"name":"College of Veterinary Medicine, Hebei Agricultural University, Baoding 071001, China"},{"name":"National Research Center of Engineering and Technology for Veterinary Biologicals, Nanjing 210014, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,9,22]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"84","DOI":"10.1016\/j.rvsc.2023.04.013","article-title":"Complete genome sequence and pathogenicity analysis of a highly pathogenic FAdV-4 strain","volume":"159","author":"Wang","year":"2023","journal-title":"Res. Vet. Sci."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"111","DOI":"10.1080\/03079457.2017.1385724","article-title":"Fowl adenovirus-induced diseases and strategies for their control\u2014A review on the current global situation","volume":"47","author":"Schachner","year":"2018","journal-title":"Avian Pathol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"247","DOI":"10.1136\/vr.124.10.247","article-title":"Hydropericarditis syndrome in broiler chickens in Pakistan","volume":"124","author":"Anjum","year":"1989","journal-title":"Vet. Rec."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"693","DOI":"10.1080\/03079459508419108","article-title":"Identification and characterization of an avian adenovirus isolated from a \u2018spiking mortality syndrome\u2019 field outbreak in broilers on the Delmarva Peninsula, USA","volume":"24","author":"Mendelson","year":"1995","journal-title":"Avian Pathol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"114","DOI":"10.1007\/s13337-013-0183-7","article-title":"Characterization of fowl adenoviruses associated with hydropericardium syndrome and inclusion body hepatitis in broiler chickens","volume":"25","author":"Mittal","year":"2014","journal-title":"Virusdisease"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"433","DOI":"10.1080\/03079459994443","article-title":"Epidemiological studies on fowl adenoviruses isolated from cases of infectious hydropericardium","volume":"28","author":"Hess","year":"1999","journal-title":"Avian Pathol."},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Grgi\u0107, H., Poljak, Z., Sharif, S., and Nagy, \u00c9. (2013). Pathogenicity and cytokine gene expression pattern of a serotype 4 fowl adenovirus isolate. PLoS ONE, 8.","DOI":"10.1371\/journal.pone.0077601"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"182","DOI":"10.1080\/03079457.2021.1872766","article-title":"Epidemiology of fowl adenovirus (FAdV) infections in South Korean chickens during 2013-2019 following introduction of FAdV-4 vaccines","volume":"50","author":"Lai","year":"2021","journal-title":"Avian Pathol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"105078","DOI":"10.1016\/j.meegid.2021.105078","article-title":"Isolation, identification and genetic characterization analysis of a fowl aviadenovirus serotype 4 strain from Tianjin, China","volume":"96","author":"Wang","year":"2021","journal-title":"Infect. Genet. Evol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"e121","DOI":"10.1111\/tbed.12691","article-title":"Epidemiological investigation of outbreaks of fowl adenovirus infections in commercial chickens in China","volume":"65","author":"Niu","year":"2018","journal-title":"Transbound. Emerg. Dis."},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Cui, J., Xu, Y., Zhou, Z., Xu, Q., Wang, J., Xiao, Y., Li, Z., and Bi, D. (2020). Pathogenicity and Molecular Typing of Fowl Adenovirus-Associated With Hepatitis\/Hydropericardium Syndrome in Central China (2015\u20132018). Front. Vet. Sci., 7.","DOI":"10.3389\/fvets.2020.00190"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"621","DOI":"10.3382\/ps\/pey153","article-title":"Chicken infectious anemia virus helps fowl adenovirus break the protection of maternal antibody and cause inclusion body hepatitis-hydropericardium syndrome in layers after using co-contaminated Newcastle disease virus-attenuated vaccine","volume":"98","author":"Su","year":"2019","journal-title":"Poult. Sci."},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Wang, K., Sun, H., Li, Y., Yang, Z., Ye, J., and Chen, H. (2019). Characterization and pathogenicity of fowl adenovirus serotype 4 isolated from eastern China. BMC Vet. Res., 15.","DOI":"10.1186\/s12917-019-2092-5"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"803","DOI":"10.3382\/ps\/pex338","article-title":"Molecular epidemiology and phylogenetic analysis of fowl adenoviruses caused hydropericardium outbreak in China during 2015","volume":"97","author":"Niu","year":"2018","journal-title":"Poult. Sci."},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Liu, A., Zhang, Y., Cui, H., Wang, X., Gao, Y., and Pan, Q. (2022). Advances in Vaccine Development of the Emerging Novel Genotype Fowl Adenovirus 4. Front. Immunol., 13.","DOI":"10.3389\/fimmu.2022.916290"},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Meng, K., Yuan, X., Yu, J., Zhang, Y., Ai, W., and Wang, Y. (2019). Identification, Pathogenicity of Novel Fowl Adenovirus Serotype 4 SDJN0105 in Shandong, China and Immunoprotective Evaluation of the Newly Developed Inactivated Oil-emulsion FAdV-4 Vaccine. Viruses, 11.","DOI":"10.3390\/v11070627"},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Zhang, Y., Liu, A., Cui, H., Qi, X., Liu, C., Zhang, Y., Li, K., Gao, L., Wang, X., and Pan, Q. (2022). An inactivated vaccine based on artificial non-pathogenic fowl adenovirus 4 protects chickens against hepatitis-hydropericardium syndrome. Vet. Microbiol., 264.","DOI":"10.1016\/j.vetmic.2021.109285"},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Lu, H., Xie, Q., Zhang, W., Zhang, J., Wang, W., Lian, M., Zhao, Z., Ren, D., Xie, S., and Lin, Y. (2022). A Novel Recombinant FAdV-4 Virus with Fiber of FAdV-8b Provides Efficient Protection against Both FAdV-4 and FAdV-8b. Viruses, 14.","DOI":"10.3390\/v14020376"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"262","DOI":"10.1016\/j.rvsc.2018.03.003","article-title":"Broad spectrum protection of broiler chickens against inclusion body hepatitis by immunizing their broiler breeder parents with a bivalent live fowl adenovirus vaccine","volume":"118","author":"Popowich","year":"2018","journal-title":"Res. Vet. Sci."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"744","DOI":"10.1016\/j.vaccine.2017.12.047","article-title":"Inactivated and live bivalent fowl adenovirus (FAdV8b + FAdV11) breeder vaccines provide broad-spectrum protection in chicks against inclusion body hepatitis (IBH)","volume":"36","author":"Gupta","year":"2018","journal-title":"Vaccine"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"396","DOI":"10.1080\/03079457.2019.1612032","article-title":"Vaccination with FAdV-8a induces protection against inclusion body hepatitis caused by homologous and heterologous strains","volume":"48","author":"Sandy","year":"2019","journal-title":"Avian Pathol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"145","DOI":"10.1016\/j.meegid.2018.03.031","article-title":"A subunit vaccine based on fiber-2 protein provides full protection against fowl adenovirus serotype 4 and induces quicker and stronger immune responses than an inactivated oil-emulsion vaccine","volume":"61","author":"Ruan","year":"2018","journal-title":"Infect. Genet. Evol."},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Wang, X., and Yuan, W. (2025). A novel recombinant chimeric Fiber-8a\/8b-AD subunit vaccine provides complete protection against both FAdV-8a and FAdV-8b. Vet. Microbiol., 303.","DOI":"10.1016\/j.vetmic.2025.110425"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"1837","DOI":"10.1016\/j.vaccine.2022.01.060","article-title":"Vaccination with a fowl adenovirus chimeric fiber protein (crecFib-4\/11) simultaneously protects chickens against hepatitis-hydropericardium syndrome (HHS) and inclusion body hepatitis (IBH)","volume":"40","author":"Schachner","year":"2022","journal-title":"Vaccine"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"70","DOI":"10.1136\/vr.140.3.70-b","article-title":"Electron microscopic demonstration of an adenovirus in the hepatocytes of birds experimentally infected with hydropericardium syndrome","volume":"140","author":"Chandra","year":"1997","journal-title":"Vet. Rec."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"99","DOI":"10.1023\/A:1005230703093","article-title":"The hydropericardium syndrome and inclusion body hepatitis in domestic fowl","volume":"32","author":"Chandra","year":"2000","journal-title":"Trop. Anim. Health Prod."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"195","DOI":"10.1080\/03079450050045440","article-title":"Detection and differentiation of avian adenoviruses: A review","volume":"29","author":"Hess","year":"2000","journal-title":"Avian Pathol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"411","DOI":"10.1016\/j.vetmic.2011.11.003","article-title":"Two fiber genes of nearly equal lengths are a common and distinctive feature of Fowl adenovirus C members","volume":"156","author":"Marek","year":"2012","journal-title":"Vet. Microbiol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"29","DOI":"10.1186\/s13567-018-0525-y","article-title":"A novel monoclonal antibody efficiently blocks the infection of serotype 4 fowl adenovirus by targeting fiber-2","volume":"49","author":"Wang","year":"2018","journal-title":"Vet. Res."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"e00954-20","DOI":"10.1128\/JVI.00954-20","article-title":"Fiber-1, Not Fiber-2, Directly Mediates the Infection of the Pathogenic Serotype 4 Fowl Adenovirus via Its Shaft and Knob Domains","volume":"94","author":"Wang","year":"2020","journal-title":"J. Virol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"33","DOI":"10.1016\/j.jim.2018.03.002","article-title":"Fiber-based fluorescent microsphere immunoassay (FMIA) as a novel multiplex serodiagnostic tool for simultaneous detection and differentiation of all clinically relevant fowl adenovirus (FAdV) serotypes","volume":"458","author":"Feichtner","year":"2018","journal-title":"J. Immunol. Methods"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"7678","DOI":"10.1128\/JVI.74.16.7678-7682.2000","article-title":"Immune response to recombinant adenovirus in humans: Capsid components from viral input are targets for vector-specific cytotoxic T lymphocytes","volume":"74","author":"Mehtali","year":"2000","journal-title":"J. Virol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"199","DOI":"10.1038\/s41426-018-0203-1","article-title":"Fiber2 and hexon genes are closely associated with the virulence of the emerging and highly pathogenic fowl adenovirus 4","volume":"7","author":"Zhang","year":"2018","journal-title":"Emerg. Microbes Infect."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"1203","DOI":"10.1016\/j.vaccine.2018.01.028","article-title":"Immunogenicity and protective efficacy of recombinant fiber-2 protein in protecting SPF chickens against fowl adenovirus 4","volume":"36","author":"Chen","year":"2018","journal-title":"Vaccine"},{"key":"ref_35","doi-asserted-by":"crossref","unstructured":"Schachner, A., De Luca, C., Heidl, S., and Hess, M. (2022). Recombinantly Expressed Chimeric Fibers Demonstrate Discrete Type-Specific Neutralizing Epitopes in the Fowl Aviadenovirus E (FAdV-E) Fiber, Promoting the Optimization of FAdV Fiber Subunit Vaccines towards Cross-Protection in vivo. Microbiol. Spectr., 10.","DOI":"10.1128\/spectrum.02123-21"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"103888","DOI":"10.1016\/j.psj.2024.103888","article-title":"A novel subunit vaccine based on Fiber1\/2 knob domain provides full protection against fowl adenovirus serotype 4 and induces stronger immune responses than a Fiber2 subunit vaccine","volume":"103","author":"Liu","year":"2024","journal-title":"Poult. Sci."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"199113","DOI":"10.1016\/j.virusres.2023.199113","article-title":"Knob domain of Fiber 2 protein provides full protection against fowl adenovirus serotype 4","volume":"330","author":"Song","year":"2023","journal-title":"Virus Res."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"73","DOI":"10.1637\/11758-100917-Reg.1","article-title":"Recombinant Fiber-2 Protein-Based Indirect ELISA for Antibody Detection of Fowl Adenovirus Serotype 4","volume":"62","author":"He","year":"2018","journal-title":"Avian Dis."},{"key":"ref_39","doi-asserted-by":"crossref","unstructured":"Zhao, K., Li, X., Lei, B., Han, Y., An, T., Zhang, W., Zhang, H., Li, B., and Yuan, W. (2023). Recombinant porcine Interferon-\u03b1 and Interleukin-2 fusion protein (rPoIFN\u03b1+IL-2) shows potent anti-pseudorabies virus activity in vitro and in vivo. Vet. Microbiol., 279.","DOI":"10.1016\/j.vetmic.2023.109678"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"143","DOI":"10.1186\/s13567-020-00869-8","article-title":"Fowl adenovirus (FAdV) fiber-based vaccine against inclusion body hepatitis (IBH) provides type-specific protection guided by humoral immunity and regulation of B and T cell response","volume":"51","author":"Schachner","year":"2020","journal-title":"Vet. Res."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"143","DOI":"10.1016\/j.vaccine.2019.10.039","article-title":"Successful reproduction of adenoviral gizzard erosion in 20-week-old SPF layer-type chickens and efficacious prophylaxis due to live vaccination with an apathogenic fowl adenovirus serotype 1 strain (CELO)","volume":"38","author":"Grafl","year":"2020","journal-title":"Vaccine"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"7225","DOI":"10.1128\/JVI.77.13.7225-7235.2003","article-title":"Flexibility of the adenovirus fiber is required for efficient receptor interaction","volume":"77","author":"Wu","year":"2003","journal-title":"J. Virol."},{"key":"ref_43","doi-asserted-by":"crossref","unstructured":"Jiao, Y., Zhao, Q., Zhao, Y., Li, Y., Pan, S., Li, Y., Liu, Y., and Shi, W. (2025). Evaluation of the Immune Effect of a Trivalent Fowl Adenovirus Inactivated Vaccine Against FAdV-4\/8a\/8b. Vet. Sci., 12.","DOI":"10.3390\/vetsci12060549"},{"key":"ref_44","doi-asserted-by":"crossref","unstructured":"Schachner, A., Gonzalez, G., Endler, L., Ito, K., and Hess, M. (2019). Fowl Adenovirus (FAdV) Recombination with Intertypic Crossovers in Genomes of FAdV-D and FAdV-E, Displaying Hybrid Serological Phenotypes. Viruses, 11.","DOI":"10.3390\/v11121094"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"573","DOI":"10.3382\/ps\/pey425","article-title":"Pathogenicity and complete genome sequence of a fowl adenovirus serotype 8b isolate from China","volume":"98","author":"Huang","year":"2019","journal-title":"Poult. Sci."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"106","DOI":"10.1080\/03079457.2015.1007919","article-title":"Chronological analysis of gross and histological lesions induced by field strains of fowl adenovirus serotypes 1, 8b and 11 in one-day-old chickens","volume":"44","author":"Steer","year":"2015","journal-title":"Avian Pathol."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"3005","DOI":"10.1007\/s00705-018-3965-9","article-title":"A comparative pathogenicity analysis of two adenovirus strains, 1\/A and 8a\/E, isolated from poultry in Poland","volume":"163","author":"Niczyporuk","year":"2018","journal-title":"Arch. Virol."}],"container-title":["Veterinary Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2306-7381\/12\/9\/920\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T18:47:23Z","timestamp":1760035643000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2306-7381\/12\/9\/920"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,9,22]]},"references-count":47,"journal-issue":{"issue":"9","published-online":{"date-parts":[[2025,9]]}},"alternative-id":["vetsci12090920"],"URL":"https:\/\/doi.org\/10.3390\/vetsci12090920","relation":{},"ISSN":["2306-7381"],"issn-type":[{"value":"2306-7381","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,9,22]]}}}