{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,23]],"date-time":"2026-04-23T05:49:56Z","timestamp":1776923396690,"version":"3.51.2"},"reference-count":51,"publisher":"MDPI AG","issue":"4","license":[{"start":{"date-parts":[[2019,12,16]],"date-time":"2019-12-16T00:00:00Z","timestamp":1576454400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"The alumni association for Kitasato University","award":["2018"],"award-info":[{"award-number":["2018"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Pathogens"],"abstract":"<jats:p>Feline coronavirus (FCoV) is a pathogen causing a lethal infectious disease in cats, feline infectious peritonitis. It has two serotypes (type I FCoV and type II FCoV). According to our previous study, type I FCoV infection is inhibited by compounds inducing intracellular cholesterol accumulation, whereas type II FCoV infection is not inhibited. Intracellular cholesterol accumulation was reported to disrupt late endosome function. Based on these findings, types I and II FCoV are considered to enter the cytosol through late and early endosomes, respectively. We investigated whether the antiviral activities of a late endosome trafficking inhibitor and cholesterol-accumulating agents are different between the FCoV serotypes. The late endosome trafficking inhibitor did not inhibit type II FCoV infection, but it inhibited type I FCoV infection. Type I FCoV infection was inhibited by cholesterol-accumulating triazoles, but not by non-cholesterol-accumulating triazoles. These phenomena were observed in both feline cell lines and feline primary macrophages. This study provides additional information on the differences in intracellular reproductive cycle between type I and type II FCoV.<\/jats:p>","DOI":"10.3390\/pathogens8040300","type":"journal-article","created":{"date-parts":[[2019,12,17]],"date-time":"2019-12-17T02:59:01Z","timestamp":1576551541000},"page":"300","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":20,"title":["Endocytic Pathway of Feline Coronavirus for Cell Entry: Differences in Serotype-Dependent Viral Entry Pathway"],"prefix":"10.3390","volume":"8","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-3468-0708","authenticated-orcid":false,"given":"Tomomi","family":"Takano","sequence":"first","affiliation":[{"name":"Laboratory of Veterinary Infectious Disease, School of Veterinary Medicine, Kitasato University, Towada, Aomori 034-8628, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yumeho","family":"Wakayama","sequence":"additional","affiliation":[{"name":"Laboratory of Veterinary Infectious Disease, School of Veterinary Medicine, Kitasato University, Towada, Aomori 034-8628, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Tomoyoshi","family":"Doki","sequence":"additional","affiliation":[{"name":"Laboratory of Veterinary Infectious Disease, School of Veterinary Medicine, Kitasato University, Towada, Aomori 034-8628, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2019,12,16]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"490","DOI":"10.1016\/j.tim.2016.03.003","article-title":"Epidemiology, genetic recombination, and pathogenesis of coronaviruses","volume":"24","author":"Su","year":"2016","journal-title":"Trends Microbiol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"469","DOI":"10.1007\/BF01718424","article-title":"Molecular cloning and sequence determination of the peplomer protein gene of feline infectious peritonitis virus type I","volume":"140","author":"Motokawa","year":"1995","journal-title":"Arch. Virol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"557","DOI":"10.1292\/jvms.54.557","article-title":"The prevalence of types I and II feline coronavirus infections in cats","volume":"54","author":"Hohdatsu","year":"1992","journal-title":"J. Vet. Med. Sci."},{"key":"ref_4","first-page":"1209","article-title":"Feline coronavirus serotypes 1 and 2: Seroprevalence and association with disease in Switzerland","volume":"12","author":"Kummrow","year":"2005","journal-title":"Clin. Diagn. Lab. Immunol."},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Wang, Y.T., Chueh, L.L., and Wan, C.H. (2014). An eight-year epidemiologic study based on baculovirus-expressed type-specific spike proteins for the differentiation of type I and II feline coronavirus infections. BMC Vet. Res., 10.","DOI":"10.1186\/s12917-014-0186-7"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"193","DOI":"10.1016\/bs.aivir.2016.08.002","article-title":"Feline coronaviruses: Pathogenesis of feline infectious peritonitis","volume":"96","author":"Tekes","year":"2016","journal-title":"Adv. Virus Res."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"123","DOI":"10.1016\/j.tvjl.2014.04.017","article-title":"An update on feline infectious peritonitis: Virology and immunopathogenesis","volume":"201","author":"Pedersen","year":"2016","journal-title":"Vet. J."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"436","DOI":"10.1128\/jvi.63.1.436-440.1989","article-title":"Intrinsic resistance of feline peritoneal macrophages to coronavirus infection correlates with in vivo virulence","volume":"63","author":"Stoddart","year":"1989","journal-title":"J. Virol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"64","DOI":"10.1016\/j.virol.2007.02.006","article-title":"TNF-alpha, produced by feline infectious peritonitis virus (FIPV)-infected macrophages, upregulates expression of type II FIPV receptor feline aminopeptidase N in feline macrophages","volume":"364","author":"Takano","year":"2007","journal-title":"Virology"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"161","DOI":"10.1016\/j.virusres.2011.03.027","article-title":"Vascular endothelial growth factor (VEGF), produced by feline infectious peritonitis (FIP) virus-infected monocytes and macrophages, induces vascular permeability and effusion in cats with FIP","volume":"158","author":"Takano","year":"2011","journal-title":"Virus Res."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"96","DOI":"10.1016\/j.antiviral.2017.07.022","article-title":"The cholesterol transport inhibitor U18666A inhibits type I feline coronavirus infection","volume":"145","author":"Takano","year":"2017","journal-title":"Antivir. Res."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"5","DOI":"10.1186\/s13567-019-0625-3","article-title":"Antiviral activity of itraconazole against type I feline coronavirus infection","volume":"50","author":"Takano","year":"2019","journal-title":"Vet. Res."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"4764","DOI":"10.1073\/pnas.0910872107","article-title":"Cholesterol trafficking is required for mTOR activation in endothelial cells","volume":"107","author":"Xu","year":"2010","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"191","DOI":"10.1016\/j.antiviral.2011.11.014","article-title":"U18666A, an intra-cellular cholesterol transport inhibitor, inhibits dengue virus entry and replication","volume":"93","author":"Poh","year":"2012","journal-title":"Antivir. Res."},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Lyu, J., Yang, E.J., and Shim, J.S. (2019). Cholesterol trafficking: An emerging therapeutic target for angiogenesis and cancer. Cells, 8.","DOI":"10.3390\/cells8050389"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"452","DOI":"10.1016\/j.chom.2013.03.006","article-title":"The antiviral effector IFITM3 disrupts intracellular cholesterol homeostasis to block viral entry","volume":"13","author":"Choi","year":"2013","journal-title":"Cell Host Microbe"},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Sobo, K., Le Blanc, I., Luyet, P.P., Fivaz, M., Ferguson, C., Parton, R.G., Gruenberg, J., and van der Goot, F.G. (2007). Late endosomal cholesterol accumulation leads to impaired intra-endosomal trafficking. PLoS ONE, 2.","DOI":"10.1371\/journal.pone.0000851"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"600","DOI":"10.1016\/j.celrep.2014.12.054","article-title":"Itraconazole inhibits enterovirus replication by targeting the oxysterol-binding protein","volume":"10","author":"Strating","year":"2015","journal-title":"Cell Rep."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"318","DOI":"10.1016\/j.redox.2018.09.003","article-title":"25-Hydroxycholesterol and 27-hydroxycholesterol inhibit human rotavirus infection by sequestering viral particles into late endosomes","volume":"19","author":"Civra","year":"2018","journal-title":"Redox Biol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"E4904","DOI":"10.1073\/pnas.1302334110","article-title":"Selective inhibitor of endosomal trafficking pathways exploited by multiple toxins and viruses","volume":"110","author":"Gillespie","year":"2013","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Vonderheit, A., and Helenius, A. (2005). Rab7 associates with early endosomes to mediate sorting and transport of Semliki forest virus to late endosomes. PLoS Biol., 3.","DOI":"10.1371\/journal.pbio.0030233"},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Saeed, M.F., Kolokoltsov, A.A., Albrecht, T., and Davey, R.A. (2010). Cellular entry of ebola virus involves uptake by a macropinocytosis-like mechanism and subsequent trafficking through early and late endosomes. PLoS Pathog., 6.","DOI":"10.1371\/journal.ppat.1001110"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"2265","DOI":"10.1111\/j.1600-0854.2008.00835.x","article-title":"Infectious adenovirus type 2 transport through early but not late endosomes","volume":"9","author":"Gastaldelli","year":"2008","journal-title":"Traffic"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"7641","DOI":"10.1128\/JVI.73.9.7641-7657.1999","article-title":"Localization of mouse hepatitis virus nonstructural proteins and RNA synthesis indicates a role for late endosomes in viral replication","volume":"73","author":"Snijder","year":"1999","journal-title":"J. Virol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"21","DOI":"10.1016\/j.virol.2008.04.046","article-title":"Lymphocytic choriomeningitis virus uses a novel endocytic pathway for infectious entry via late endosomes","volume":"378","author":"Quirin","year":"2008","journal-title":"Virology"},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Burkard, C., Verheije, M.H., Wicht, O., van Kasteren, S.I., van Kuppeveld, F.J., Haagmans, B.L., Pelkmans, L., Rottier, P.J., Bosch, B.J., and de Haan, C.A. (2014). Coronavirus cell entry occurs through the endo-\/lysosomal pathway in a proteolysis-dependent manner. PLoS Pathog., 10.","DOI":"10.1371\/journal.ppat.1004502"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"142","DOI":"10.1165\/rcmb.2007-0217OC","article-title":"Amiodarone alters late endosomes and inhibits SARS coronavirus infection at a post-endosomal level","volume":"39","author":"Stadler","year":"2008","journal-title":"Am. J. Respir. Cell Mol. Biol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"1011","DOI":"10.3390\/v4061011","article-title":"Mechanisms of coronavirus cell entry mediated by the viral spike protein","volume":"4","author":"Belouzard","year":"2012","journal-title":"Viruses"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"6048","DOI":"10.1128\/JVI.78.11.6048-6054.2004","article-title":"Cleavage inhibition of the murine coronavirus spike protein by a furin-like enzyme affects cell-cell but not virus-cell fusion","volume":"78","author":"Stadler","year":"2004","journal-title":"J. Virol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"4240","DOI":"10.1073\/pnas.0306446101","article-title":"Characterization of severe acute respiratory syndrome-associated coronavirus (SARS-CoV) spike glycoprotein-mediated viral entry","volume":"101","author":"Simmons","year":"2004","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"8887","DOI":"10.1128\/JVI.00415-08","article-title":"Cathepsin L functionally cleaves the severe acute respiratory syndrome coronavirus class I fusion protein upstream of rather than adjacent to the fusion peptide","volume":"82","author":"Bosch","year":"2008","journal-title":"J. Virol."},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Qian, Z., Dominguez, S.R., and Holmes, K.V. (2013). Role of the spike glycoprotein of human Middle East respiratory syndrome coronavirus (MERS-CoV) in virus entry and syncytia formation. PLoS ONE, 8.","DOI":"10.1371\/journal.pone.0076469"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"605","DOI":"10.1016\/j.antiviral.2013.09.028","article-title":"Proteolytic activation of the SARS-coronavirus spike protein: Cutting enzymes at the cutting edge of antiviral research","volume":"100","author":"Simmons","year":"2013","journal-title":"Antivir. Res."},{"key":"ref_34","doi-asserted-by":"crossref","unstructured":"Cermak, S., Kosicek, M., Mladenovic-Djordjevic, A., Smiljanic, K., Kanazir, S., and Hecimovic, S. (2016). Loss of Cathepsin B and l leads to lysosomal dysfunction, NPC-like cholesterol sequestration and accumulation of the key Alzheimer\u2019s proteins. PLoS ONE, 11.","DOI":"10.1371\/journal.pone.0167428"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"235","DOI":"10.1016\/j.vetmic.2008.05.019","article-title":"Differential role for low pH and cathepsin-mediated cleavage of the viral spike protein during entry of serotype II feline coronaviruses","volume":"132","author":"Regan","year":"2008","journal-title":"Vet. Microbiol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"92","DOI":"10.1016\/j.immuni.2012.11.005","article-title":"Interferon-inducible cholesterol-25-hydroxylase broadly inhibits viral entry by production of 25-hydroxycholesterol","volume":"38","author":"Liu","year":"2013","journal-title":"Immunity"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"446","DOI":"10.1016\/j.immuni.2017.02.012","article-title":"25-Hydroxycholesterol protects host against Zika virus infection and its associated microcephaly in a mouse model","volume":"46","author":"Li","year":"2017","journal-title":"Immunity"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"7487","DOI":"10.1038\/srep07487","article-title":"Inhibition of pathogenic non-enveloped viruses by 25-hydroxycholesterol and 27-hydroxycholesterol","volume":"4","author":"Civra","year":"2014","journal-title":"Sci. Rep."},{"key":"ref_39","doi-asserted-by":"crossref","unstructured":"Shrivastava-Ranjan, P., Bergeron, \u00c9., Chakrabarti, A.K., Albari\u00f1o, C.G., Flint, M., Nichol, S.T., and Spiropoulou, C.F. (2016). 25-Hydroxycholesterol inhibition of Lassa virus infection through aberrant GP1 glycosylation. MBio, 7.","DOI":"10.1128\/mBio.01808-16"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"395","DOI":"10.3851\/IMP3232","article-title":"Antiviral effect of 25-hydroxycholesterol against porcine reproductive and respiratory syndrome virus in vitro","volume":"23","author":"Dong","year":"2018","journal-title":"Antivir. Ther."},{"key":"ref_41","doi-asserted-by":"crossref","unstructured":"Shawli, G.T., Adeyemi, O.O., Stonehouse, N.J., and Herod, M.R. (2019). The Oxysterol 25-Hydroxycholesterol Inhibits Replication of Murine Norovirus. Viruses, 11.","DOI":"10.3390\/v11020097"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"497","DOI":"10.1002\/jsfa.2740550402","article-title":"Cholesterol oxides and atherosclerosis: A review","volume":"55","author":"Kumar","year":"1991","journal-title":"J. Sci. Food Agric."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"2045","DOI":"10.1253\/circj.CJ-10-0860","article-title":"Macrophages, oxysterols and atherosclerosis","volume":"74","author":"Shibata","year":"2010","journal-title":"Circ. J."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"1025","DOI":"10.1099\/vir.0.83558-0","article-title":"Analysis of the mechanism of antibody-dependent enhancement of feline infectious peritonitis virus infection: Aminopeptidase N is not important and a process of acidification of the endosome is necessary","volume":"89","author":"Takano","year":"2008","journal-title":"J. Gen. Virol."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"89","DOI":"10.1073\/pnas.1619571114","article-title":"Triazoles inhibit cholesterol export from lysosomes by binding to NPC1","volume":"114","author":"Trinh","year":"2017","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"1213","DOI":"10.1016\/j.cell.2009.03.049","article-title":"Structure of N-terminal domain of NPC1 reveals distinct subdomains for binding and transfer of cholesterol","volume":"137","author":"Kwon","year":"2009","journal-title":"Cell"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"174","DOI":"10.1021\/acschembio.6b00849","article-title":"Simultaneous targeting of NPC1 and VDAC1 by itraconazole leads to synergistic inhibition of mTOR signaling and angiogenesis","volume":"12","author":"Head","year":"2016","journal-title":"ACS Chem. Biol."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"125","DOI":"10.1007\/s00705-015-2655-0","article-title":"Differential effect of cholesterol on type I and II feline coronavirus infection","volume":"161","author":"Takano","year":"2016","journal-title":"Arch. Virol."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.virol.2018.08.022","article-title":"Characterizing replication kinetics and plaque production of type I feline infectious peritonitis virus in three feline cell lines","volume":"525","author":"Mettelman","year":"2018","journal-title":"Virology"},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"226","DOI":"10.1016\/j.virol.2019.08.030","article-title":"Generating and evaluating type I interferon receptor-deficient and feline TMPRSS2-expressing cells for propagating serotype I feline infectious peritonitis virus","volume":"537","author":"Mettelman","year":"2019","journal-title":"Virology"},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"13","DOI":"10.1016\/0378-1135(91)90096-X","article-title":"Antigenic analysis of feline coronaviruses with monoclonal antibodies (MAbs): Preparation of MAbs which discriminate between FIPV strain 79-1146 and FECV strain 79-1683","volume":"28","author":"Hohdatsu","year":"1991","journal-title":"Vet. Microbiol."}],"container-title":["Pathogens"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-0817\/8\/4\/300\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T13:42:48Z","timestamp":1760190168000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-0817\/8\/4\/300"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2019,12,16]]},"references-count":51,"journal-issue":{"issue":"4","published-online":{"date-parts":[[2019,12]]}},"alternative-id":["pathogens8040300"],"URL":"https:\/\/doi.org\/10.3390\/pathogens8040300","relation":{},"ISSN":["2076-0817"],"issn-type":[{"value":"2076-0817","type":"electronic"}],"subject":[],"published":{"date-parts":[[2019,12,16]]}}}