{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,28]],"date-time":"2026-01-28T03:06:20Z","timestamp":1769569580203,"version":"3.49.0"},"reference-count":37,"publisher":"MDPI AG","issue":"6","license":[{"start":{"date-parts":[[2019,6,3]],"date-time":"2019-06-03T00:00:00Z","timestamp":1559520000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100002999","name":"Ministerie van Volksgezondheid, Welzijn en Sport","doi-asserted-by":"publisher","award":["SPR S\/112008"],"award-info":[{"award-number":["SPR S\/112008"]}],"id":[{"id":"10.13039\/501100002999","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Viruses"],"abstract":"<jats:p>Human respiratory syncytial virus (RSV) is a major cause of severe lower respiratory tract disease requiring hospitalization in infants. There are no market-approved vaccines or antiviral agents available, but a growing number of vaccines and therapeutics are in (pre)clinical stages of development. Reliable animal models are crucial to evaluate new vaccine concepts, but in vivo RSV research is hampered by the lack of well-characterized animal models that faithfully mimic the pathogenesis of RSV infection in humans. Mice are frequently used in RSV infection and vaccination studies. However, differences in the use of mouse strains, RSV subtypes, and methodology often lead to divergent study outcomes. To our knowledge, a comparison between different RSV inoculation methods in mice has not been described in the literature, even though multiple methods are being used across different studies. In this study, we evaluated various pathological and immunological parameters in BALB\/c mice after intratracheal or intranasal inoculation with RSV-A2. Our study reveals that intranasal inoculation induces robust pathology and inflammation, whereas this is not the case for intratracheal inoculation. As immunopathology is an important characteristic of RSV disease in infants, these data suggest that in mice intranasal inoculation is a more appropriate method to study RSV infection than intratracheal inoculation. These findings will contribute to the rational experimental design of future in vivo RSV experiments.<\/jats:p>","DOI":"10.3390\/v11060508","type":"journal-article","created":{"date-parts":[[2019,6,3]],"date-time":"2019-06-03T11:31:01Z","timestamp":1559561461000},"page":"508","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":9,"title":["Pathogenesis of Respiratory Syncytial Virus Infection in BALB\/c Mice Differs Between Intratracheal and Intranasal Inoculation"],"prefix":"10.3390","volume":"11","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-1541-9053","authenticated-orcid":false,"given":"Elisabeth A.","family":"van Erp","sequence":"first","affiliation":[{"name":"Centre for Infectious Disease Control, National Institute for Public Health and the Environment (RIVM), 3721 MA Bilthoven, The Netherlands"},{"name":"Section Pediatric Infectious Diseases, Laboratory of Medical Immunology, Radboud Institute for Molecular Life Sciences, Radboudumc, 6525 GA Nijmegen, The Netherlands"},{"name":"Radboud Center for Infectious Diseases, Radboudumc, 6525 GA Nijmegen, The Netherlands"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4386-3995","authenticated-orcid":false,"given":"Anke J.","family":"Lakerveld","sequence":"additional","affiliation":[{"name":"Centre for Infectious Disease Control, National Institute for Public Health and the Environment (RIVM), 3721 MA Bilthoven, The Netherlands"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"H. Lie","family":"Mulder","sequence":"additional","affiliation":[{"name":"Centre for Infectious Disease Control, National Institute for Public Health and the Environment (RIVM), 3721 MA Bilthoven, The Netherlands"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Willem","family":"Luytjes","sequence":"additional","affiliation":[{"name":"Centre for Infectious Disease Control, National Institute for Public Health and the Environment (RIVM), 3721 MA Bilthoven, The Netherlands"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Gerben","family":"Ferwerda","sequence":"additional","affiliation":[{"name":"Section Pediatric Infectious Diseases, Laboratory of Medical Immunology, Radboud Institute for Molecular Life Sciences, Radboudumc, 6525 GA Nijmegen, The Netherlands"},{"name":"Radboud Center for Infectious Diseases, Radboudumc, 6525 GA Nijmegen, The Netherlands"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4369-6055","authenticated-orcid":false,"given":"Puck B.","family":"van Kasteren","sequence":"additional","affiliation":[{"name":"Centre for Infectious Disease Control, National Institute for Public Health and the Environment (RIVM), 3721 MA Bilthoven, The Netherlands"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2019,6,3]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"946","DOI":"10.1016\/S0140-6736(17)30938-8","article-title":"Global, regional, and national disease burden estimates of acute lower respiratory infections due to respiratory syncytial virus in young children in 2015: a systematic review and modelling study","volume":"390","author":"Shi","year":"2017","journal-title":"Lancet"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"L148","DOI":"10.1152\/ajplung.00065.2011","article-title":"Animal models of human respiratory syncytial virus disease","volume":"301","author":"Bem","year":"2011","journal-title":"Am. J. Physiology. Lung Cell. Mol. Physiol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"469","DOI":"10.1016\/j.vaccine.2016.11.054","article-title":"Animal models of respiratory syncytial virus infection","volume":"35","author":"Taylor","year":"2017","journal-title":"Vaccine"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1856","DOI":"10.1086\/386372","article-title":"Respiratory syncytial virus induces pneumonia, cytokine response, airway obstruction, and chronic inflammatory infiltrates associated with long-term airway hyperresponsiveness in mice","volume":"189","author":"Jafri","year":"2004","journal-title":"J. Infect. Dis."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"149","DOI":"10.1111\/j.1600-065X.2010.00972.x","article-title":"Biological challenges and technological opportunities for respiratory syncytial virus vaccine development","volume":"239","author":"Graham","year":"2011","journal-title":"Immunol. Rev."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"470","DOI":"10.1111\/j.1365-2249.2005.02780.x","article-title":"Respiratory syncytial virus and neutrophil activation","volume":"140","author":"Bataki","year":"2005","journal-title":"Clin. Exp. Immunol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"764","DOI":"10.1128\/iai.26.2.764-766.1979","article-title":"Respiratory syncytial virus infection in inbred mice","volume":"26","author":"Prince","year":"1979","journal-title":"Infect. Immun."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"92","DOI":"10.1002\/jmv.2196","article-title":"Genetic susceptibility to respiratory syncytial virus infection in inbred mice","volume":"67","author":"Stark","year":"2002","journal-title":"J. Med. Virol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"5782","DOI":"10.1128\/JVI.01693-10","article-title":"Differential pathogenesis of respiratory syncytial virus clinical isolates in BALB\/c mice","volume":"85","author":"Stokes","year":"2011","journal-title":"J. Virol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"977","DOI":"10.2353\/ajpath.2006.051055","article-title":"Differential immune responses and pulmonary pathophysiology are induced by two different strains of respiratory syncytial virus","volume":"169","author":"Lukacs","year":"2006","journal-title":"Am. J. Pathol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"649","DOI":"10.1128\/iai.43.2.649-655.1984","article-title":"Respiratory syncytial virus infection in mice","volume":"43","author":"Taylor","year":"1984","journal-title":"Infect. Immun."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"10710","DOI":"10.1128\/JVI.00986-09","article-title":"Respiratory syncytial virus grown in Vero cells contains a truncated attachment protein that alters its infectivity and dependence on glycosaminoglycans","volume":"83","author":"Kwilas","year":"2009","journal-title":"J. Virol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1415","DOI":"10.1016\/0264-410X(93)90170-3","article-title":"Enhanced pulmonary pathology associated with the use of formalin-inactivated respiratory syncytial virus vaccine in cotton rats is not a unique viral phenomenon","volume":"11","author":"Piedra","year":"1993","journal-title":"Vaccine"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"30","DOI":"10.1016\/j.ajpath.2011.03.026","article-title":"Pathogenesis of Influenza A\/H5N1 virus infection in ferrets differs between intranasal and intratracheal routes of inoculation","volume":"179","author":"Bodewes","year":"2011","journal-title":"Am. J. Pathol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"1534","DOI":"10.1126\/science.1213362","article-title":"Airborne transmission of influenza A\/H5N1 virus between ferrets","volume":"336","author":"Herfst","year":"2012","journal-title":"Science"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"127","DOI":"10.1078\/0940-2993-00240","article-title":"Pulmonary microbial infection in mice: comparison of different application methods and correlation of bacterial numbers and histopathology","volume":"54","author":"Munder","year":"2002","journal-title":"Exp. Toxicol. Pathol. Off. J. Ges. Fur Toxikol. Pathol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"148","DOI":"10.1186\/1465-9921-12-148","article-title":"Acute intratracheal Pseudomonas aeruginosa infection in cystic fibrosis mice is age-independent","volume":"12","author":"Munder","year":"2011","journal-title":"Respir. Res."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"209","DOI":"10.1007\/978-1-4939-3687-8_15","article-title":"Mouse and Cotton Rat Models of Human Respiratory Syncytial Virus","volume":"1442","author":"Rudd","year":"2016","journal-title":"Methods Mol. Biol. (Cliftonn. J.)"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1015","DOI":"10.1172\/JCI5849","article-title":"Surfactant protein-A enhances respiratory syncytial virus clearance in vivo","volume":"103","author":"LeVine","year":"1999","journal-title":"J. Clin. Investig."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"310","DOI":"10.3892\/mmr.2017.6558","article-title":"Respiratory syncytial virus infection accelerates lung fibrosis through the unfolded protein response in a bleomycin-induced pulmonary fibrosis animal model","volume":"16","author":"Wang","year":"2017","journal-title":"Mol. Med. Rep."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"331","DOI":"10.1093\/toxsci\/kfg031","article-title":"Effect of preexposure to ultrafine carbon black on respiratory syncytial virus infection in mice","volume":"72","author":"Lambert","year":"2003","journal-title":"Toxicol. Sci. Off. J. Soc. Toxicol."},{"key":"ref_22","unstructured":"Mahy, B.W.J., and Kangro, H.O. (1996). Virus Isolation and Quantitation. Virology Methods Manual, Academic Press."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"L231","DOI":"10.1152\/ajplung.00049.2003","article-title":"Evaluation of lung injury in rats and mice","volume":"286","author":"Parker","year":"2004","journal-title":"Am. J. Physiol. Lung Cell. Mol. Physiol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"L833","DOI":"10.1152\/ajplung.00173.2001","article-title":"Distribution of intranasal instillations in mice: effects of volume, time, body position, and anesthesia","volume":"282","author":"Southam","year":"2002","journal-title":"Am. J. Physiol. Lung Cell. Mol. Physiol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"369","DOI":"10.1165\/ajrcmb\/6.4.369","article-title":"Respiratory syncytial virus infection of human primary nasal and bronchial epithelial cell cultures and bronchoalveolar macrophages","volume":"6","author":"Becker","year":"1992","journal-title":"Am. J. Respir. Cell Mol. Biol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"842","DOI":"10.1164\/rccm.201304-0750OC","article-title":"Relative respiratory syncytial virus cytopathogenesis in upper and lower respiratory tract epithelium","volume":"188","author":"Canning","year":"2013","journal-title":"Am. J. Respir. Crit. Care Med."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"272","DOI":"10.1258\/002367704323133655","article-title":"Influence of different anaesthetics on pro-inflammatory cytokine expression in rat spleen","volume":"38","author":"Bette","year":"2004","journal-title":"Lab. Anim."},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Chiang, N., Schwab, J.M., Fredman, G., Kasuga, K., Gelman, S., and Serhan, C.N. (2008). Anesthetics impact the resolution of inflammation. PLoS ONE, 3.","DOI":"10.1371\/journal.pone.0001879"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"241","DOI":"10.2174\/1389557053175407","article-title":"Anti-inflammatory effects of intravenous anesthetics on endotoxemia","volume":"5","author":"Taniguchi","year":"2005","journal-title":"Mini Rev. Med. Chem."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"67","DOI":"10.1186\/s13054-017-1645-x","article-title":"Anti-inflammatory properties of anesthetic agents","volume":"21","author":"Cruz","year":"2017","journal-title":"Crit. Care (Lond. Engl.)"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"485","DOI":"10.1097\/SHK.0b013e3181c3cea5","article-title":"Lipoteichoic acid-induced TNF-alpha and IL-6 gene expressions and oxidative stress production in macrophages are suppressed by ketamine through downregulating Toll-like receptor 2-mediated activation oF ERK1\/2 and NFkappaB","volume":"33","author":"Chang","year":"2010","journal-title":"Shock (Augustaga.)"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"1051","DOI":"10.1213\/ANE.0b013e3181ed12fc","article-title":"The effect of ketamine anesthesia on the immune function of mice with postoperative septicemia","volume":"111","author":"Takahashi","year":"2010","journal-title":"Anesth. Analg."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"1750","DOI":"10.1128\/AAC.36.8.1750","article-title":"Inhibition of rabies virus transcription in rat cortical neurons with the dissociative anesthetic ketamine","volume":"36","author":"Lockhart","year":"1992","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"143","DOI":"10.1213\/ANE.0000000000001296","article-title":"Inhaled Anesthetics Exert Different Protective Properties in a Mouse Model of Ventilator-Induced Lung Injury","volume":"123","author":"Strosing","year":"2016","journal-title":"Anesth. Analg."},{"key":"ref_35","first-page":"402","article-title":"From the Cover: Prolonged Exposure to Volatile Anesthetic Isoflurane Worsens the Outcome of Polymicrobial Abdominal Sepsis","volume":"156","author":"Koutsogiannaki","year":"2017","journal-title":"Toxicol. Sci. Off. J. Soc. Toxicol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"1603","DOI":"10.4049\/jimmunol.121.4.1603","article-title":"Quantitative differences in immune responses during the various stages of the estrous cycle in female BALB\/c mice","volume":"121","author":"Krzych","year":"1978","journal-title":"J. Immunol."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"16","DOI":"10.1016\/j.virol.2004.06.051","article-title":"Myocarditis susceptibility in female mice depends upon ovarian cycle phase at infection","volume":"330","author":"Schwartz","year":"2004","journal-title":"Virology"}],"container-title":["Viruses"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1999-4915\/11\/6\/508\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T12:55:43Z","timestamp":1760187343000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1999-4915\/11\/6\/508"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2019,6,3]]},"references-count":37,"journal-issue":{"issue":"6","published-online":{"date-parts":[[2019,6]]}},"alternative-id":["v11060508"],"URL":"https:\/\/doi.org\/10.3390\/v11060508","relation":{},"ISSN":["1999-4915"],"issn-type":[{"value":"1999-4915","type":"electronic"}],"subject":[],"published":{"date-parts":[[2019,6,3]]}}}