{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,17]],"date-time":"2026-01-17T23:14:03Z","timestamp":1768691643730,"version":"3.49.0"},"reference-count":28,"publisher":"MDPI AG","issue":"10","license":[{"start":{"date-parts":[[2023,10,10]],"date-time":"2023-10-10T00:00:00Z","timestamp":1696896000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"National Key Research and Development Program of China","award":["2022YFD1800602"],"award-info":[{"award-number":["2022YFD1800602"]}]},{"name":"National Key Research and Development Program of China","award":["GY202104"],"award-info":[{"award-number":["GY202104"]}]},{"name":"National Key Research and Development Program of China","award":["202105"],"award-info":[{"award-number":["202105"]}]},{"name":"Public Welfare Project of the China Institute of Veterinary Drug Control","award":["2022YFD1800602"],"award-info":[{"award-number":["2022YFD1800602"]}]},{"name":"Public Welfare Project of the China Institute of Veterinary Drug Control","award":["GY202104"],"award-info":[{"award-number":["GY202104"]}]},{"name":"Public Welfare Project of the China Institute of Veterinary Drug Control","award":["202105"],"award-info":[{"award-number":["202105"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Viruses"],"abstract":"<jats:p>Single or mixed infections of multiple pathogens such as avian hepatitis E virus (aHEV) and avian leukosis virus subgroup J (ALV-J) have been detected in numerous laying hens with severe liver injury in China. Thus, aHEV and immunosuppressive viruses are speculated to cause co-infections. In this study, co-infection with aHEV and fowl adenovirus (FAdV) was confirmed by nested RT-PCR and recombinase-aided amplification combined with gene sequencing in two flocks with severe liver injury. Subsequently, the two reference strains, aHEV and FAdV-4, were inoculated into LMH cells to identify their co-infection potential. Confocal microscopy revealed aHEV and FAdV-4 co-infected LMH cells. In addition, the replication dynamics of aHEV and FAdV-4 along with the expression levels of immuno-cytokines were measured. The results indicated colocalization of aHEV and FAdV-4 and inhibition of viral replication in LMH cells. The transcription levels of MDA5, Mx, OASL, and IFN-\u03b1 were significantly upregulated in LMH cells, whereas those of immune-related factors induced by FAdV-4 were downregulated upon FAdV-4 and aHEV co-infection. These results confirmed the co-infection of aHEV and FAdV-4 in vitro and prompted the antagonistic pathogenic effects of FAdV-4 and aHEV, thereby providing novel insights into the counterbalancing effects of these viruses.<\/jats:p>","DOI":"10.3390\/v15102072","type":"journal-article","created":{"date-parts":[[2023,10,10]],"date-time":"2023-10-10T06:14:04Z","timestamp":1696918444000},"page":"2072","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["FAdV-4 Promotes Expression of Multiple Cytokines and Inhibits the Proliferation of aHEV in LMH Cells"],"prefix":"10.3390","volume":"15","author":[{"given":"Lidan","family":"Hou","sequence":"first","affiliation":[{"name":"China Institute of Veterinary Drug Control, Beijing 100081, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Wei","family":"Wang","sequence":"additional","affiliation":[{"name":"Zhaoyuan Center for Disease Control and Prevention, Yantai 265400, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Zengna","family":"Chi","sequence":"additional","affiliation":[{"name":"College of Animal Science and Veterinary Medicine, Shandong Agricultural University, Taian 271018, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yawen","family":"Zhang","sequence":"additional","affiliation":[{"name":"College of Animal Science and Veterinary Medicine, Shandong Agricultural University, Taian 271018, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Zhong","family":"Zou","sequence":"additional","affiliation":[{"name":"Hubei Jiangxia Laboratory, Wuhan 430200, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Peng","family":"Zhao","sequence":"additional","affiliation":[{"name":"College of Animal Science and Veterinary Medicine, Shandong Agricultural University, Taian 271018, China"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2023,10,10]]},"reference":[{"key":"ref_1","first-page":"500","article-title":"British Columbia. \u201cHepatitis-splenomegaly\u201d syndrome in commercial egg laying hens","volume":"32","author":"Ritchie","year":"1991","journal-title":"Can. Vet J."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"725","DOI":"10.1080\/03079459408419041","article-title":"Necrotic, haemorrhagic, hepatomegalic hepatitis associated with vasculitis and amyloidosis in commercial laying hens","volume":"23","author":"Tablante","year":"1994","journal-title":"Avian Pathol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"4197","DOI":"10.1128\/JCM.40.11.4197-4202.2002","article-title":"Heterogeneity and seroprevalence of a newly identified avian hepatitis e virus from chickens in the United States","volume":"40","author":"Huang","year":"2002","journal-title":"J. Clin. Microbiol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"31","DOI":"10.1016\/j.vetmic.2008.12.010","article-title":"Evidence of widespread infection of avian hepatitis E virus (avian HEV) in chickens from Spain","volume":"137","author":"Peralta","year":"2009","journal-title":"Vet. Microbiol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"863","DOI":"10.1099\/vir.0.007179-0","article-title":"Sequence analysis and comparison of avian hepatitis E viruses from Australia and Europe indicate the existence of different genotypes","volume":"90","author":"Bilic","year":"2009","journal-title":"J. Gen. Virol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1469","DOI":"10.3201\/eid1609.100626","article-title":"Analysis of avian hepatitis E virus from chickens, China","volume":"16","author":"Zhao","year":"2010","journal-title":"Emerg. Infect. Dis."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1365","DOI":"10.3201\/eid1808.111669","article-title":"Putative novel genotype of avian hepatitis E virus, Hungary, 2010","volume":"18","author":"Banyai","year":"2012","journal-title":"Emerg. Infect. Dis."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1113","DOI":"10.3382\/ps.2011-02050","article-title":"Outbreak of gizzard erosion associated with fowl adenovirus infection in Korea","volume":"91","author":"Lim","year":"2012","journal-title":"Poult. Sci."},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Zhang, X.L., Li, W.F., Yuan, S., Guo, J.Y., Li, Z.L., Chi, S.H., Huang, W.J., Li, X.W., Huang, S.J., and Shao, J.W. (2019). Meta-transcriptomic analysis reveals a new subtype of genotype 3 avian hepatitis E virus in chicken flocks with high mortality in Guangdong, China. BMC Vet. Res., 15.","DOI":"10.1186\/s12917-019-1884-y"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"2403","DOI":"10.1111\/tbed.13578","article-title":"Genetic diversity of avian hepatitis E virus in China, 2018\u20132019","volume":"67","author":"Su","year":"2020","journal-title":"Transbound. Emerg. Dis."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1287","DOI":"10.1016\/j.psj.2019.10.067","article-title":"Co-infection with avian hepatitis E virus and avian leukosis virus subgroup J as the cause of an outbreak of hepatitis and liver hemorrhagic syndromes in a brown layer chicken flock in China","volume":"99","author":"Sun","year":"2020","journal-title":"Poult. Sci."},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Sun, Y., Du, T., Liu, B., Syed, S.F., Chen, Y., Li, H., Wang, X., Zhang, G., Zhou, E.M., and Zhao, Q. (2016). Seroprevalence of avian hepatitis E virus and avian leucosis virus subgroup J in chicken flocks with hepatitis syndrome, China. BMC Vet. Res., 12.","DOI":"10.1186\/s12917-016-0892-4"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"698","DOI":"10.1637\/11386-013016-Reg.1","article-title":"Natural Infection with Avian Hepatitis E Virus and Marek\u2019s Disease Virus in Brown Layer Chickens in China","volume":"60","author":"Yang","year":"2016","journal-title":"Avian Dis."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"776","DOI":"10.1111\/tbed.13082","article-title":"Epidemiological investigation of the novel genotype avian hepatitis E virus and co-infected immunosuppressive viruses in farms with hepatic rupture haemorrhage syndrome, recently emerged in China","volume":"66","author":"Su","year":"2019","journal-title":"Transbound. Emerg. Dis."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"1699","DOI":"10.3382\/ps\/pex414","article-title":"Co-infection of fowl adenovirus with different immunosuppressive viruses in a chicken flock","volume":"97","author":"Meng","year":"2018","journal-title":"Poult. Sci."},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Yu, G., Lin, Y., Dou, Y., Tang, Y., and Diao, Y. (2019). Prevalence of Fowl Adenovirus Serotype 4 and Co-Infection by Immunosuppressive Viruses in Fowl with Hydropericardium Hepatitis Syndrome in Shandong Province, China. Viruses, 11.","DOI":"10.3390\/v11060517"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"101540","DOI":"10.1016\/j.psj.2021.101540","article-title":"Development of a PCR-based dot blot assay for the detection of fowl adenovirus","volume":"101","author":"Hou","year":"2022","journal-title":"Poult. Sci."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"693","DOI":"10.1099\/vir.0.19582-0","article-title":"Genetic identification of avian hepatitis E virus (HEV) from healthy chicken flocks and characterization of the capsid gene of 14 avian HEV isolates from chickens with hepatitis-splenomegaly syndrome in different geographical regions of the United States","volume":"85","author":"Sun","year":"2004","journal-title":"J. Gen. Virol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"102591","DOI":"10.1016\/j.psj.2023.102591","article-title":"Combined inhibitory effect of interferon and antiserum on avian hepatitis E virus","volume":"102","author":"Yan","year":"2023","journal-title":"Poult. Sci."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"1339","DOI":"10.1128\/JCM.01626-10","article-title":"TaqMan real-time reverse transcription-PCR assay for universal detection and quantification of avian hepatitis E virus from clinical samples in the presence of a heterologous internal control RNA","volume":"49","author":"Troxler","year":"2011","journal-title":"J. Clin. Microbiol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"21","DOI":"10.1016\/j.vetimm.2016.08.013","article-title":"Marek\u2019s disease virus immunosuppression alters host cellular responses and immune gene expression in the skin of infected chickens","volume":"180","author":"Heidari","year":"2016","journal-title":"Vet. Immunol. Immunopathol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"57","DOI":"10.1007\/s12250-015-3696-y","article-title":"Responses of the Toll-like receptor and melanoma differentiation-associated protein 5 signaling pathways to avian infectious bronchitis virus infection in chicks","volume":"31","author":"He","year":"2016","journal-title":"Virol. Sin."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"76","DOI":"10.1038\/s41426-018-0075-4","article-title":"Avian hepatitis E virus infection of duck, goose, and rabbit in northwest China","volume":"7","author":"Liu","year":"2018","journal-title":"Emerg. Microbes Infect."},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Liu, B., Chen, Y., Zhao, L., Zhang, M., Ren, X., Zhang, Y., Zhang, B., Fan, M., Zhao, Q., and Zhou, E.M. (2020). Identification and pathogenicity of a novel genotype avian hepatitis E virus from silkie fowl (gallus gallus). Vet. Microbiol., 245.","DOI":"10.1016\/j.vetmic.2020.108688"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"6657","DOI":"10.1016\/j.psj.2020.09.083","article-title":"Research Note: A putative novel subtype of the avian hepatitis E virus of genotype 3, Jiangxi province, China","volume":"99","author":"Li","year":"2020","journal-title":"Poult. Sci."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"281","DOI":"10.1007\/s00705-021-05318-1","article-title":"The pros and cons of cytokines for fowl adenovirus serotype 4 infection","volume":"167","author":"Wang","year":"2022","journal-title":"Arch. Virol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"65","DOI":"10.1016\/j.vetmic.2017.12.007","article-title":"Insights into leghorn male hepatocellular cells response to fowl adenovirus serotype 4 infection by transcriptome analysis","volume":"214","author":"Zhang","year":"2018","journal-title":"Vet. Microbiol."},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Wang, Z., and Zhao, J. (2019). Pathogenesis of Hypervirulent Fowl Adenovirus Serotype 4: The Contributions of Viral and Host Factors. Viruses, 11.","DOI":"10.3390\/v11080741"}],"container-title":["Viruses"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1999-4915\/15\/10\/2072\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T21:04:16Z","timestamp":1760130256000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1999-4915\/15\/10\/2072"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,10,10]]},"references-count":28,"journal-issue":{"issue":"10","published-online":{"date-parts":[[2023,10]]}},"alternative-id":["v15102072"],"URL":"https:\/\/doi.org\/10.3390\/v15102072","relation":{},"ISSN":["1999-4915"],"issn-type":[{"value":"1999-4915","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,10,10]]}}}