{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,24]],"date-time":"2026-09-24T02:48:05Z","timestamp":1790218085776,"version":"4.1.0"},"reference-count":74,"publisher":"American Society for Microbiology","issue":"4","license":[{"start":{"date-parts":[[2024,4,10]],"date-time":"2024-04-10T00:00:00Z","timestamp":1712707200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2024,4,10]],"date-time":"2024-04-10T00:00:00Z","timestamp":1712707200000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"funder":[{"DOI":"10.13039\/501100000691","name":"Academy of Medical Sciences","doi-asserted-by":"crossref","award":["SBF006\\1024"],"award-info":[{"award-number":["SBF006\\1024"]}],"id":[{"id":"10.13039\/501100000691","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/100010269","name":"Wellcome Trust","doi-asserted-by":"crossref","award":["WT105618MA"],"award-info":[{"award-number":["WT105618MA"]}],"id":[{"id":"10.13039\/100010269","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/100014013","name":"UK Research and Innovation","doi-asserted-by":"crossref","award":["MR\/N006364\/2"],"award-info":[{"award-number":["MR\/N006364\/2"]}],"id":[{"id":"10.13039\/100014013","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/100014013","name":"UK Research and Innovation","doi-asserted-by":"crossref","award":["MR\/V033417\/1"],"award-info":[{"award-number":["MR\/V033417\/1"]}],"id":[{"id":"10.13039\/100014013","id-type":"DOI","asserted-by":"crossref"}]},{"name":"Alzheimer's Research UK","award":["ARUK-NC2021-SW"],"award-info":[{"award-number":["ARUK-NC2021-SW"]}]},{"name":"HHS | National Institutes of Health","award":["AI052733-16"],"award-info":[{"award-number":["AI052733-16"]}]},{"name":"HHS | National Institutes of Health","award":["AI152078-01"],"award-info":[{"award-number":["AI152078-01"]}]},{"name":"HHS | National Institutes of Health","award":["HL059842 19"],"award-info":[{"award-number":["HL059842 19"]}]},{"name":"National Science Foundation","award":["NS127076"],"award-info":[{"award-number":["NS127076"]}]},{"name":"HHS | National Institutes of Health","award":["AI168539"],"award-info":[{"award-number":["AI168539"]}]},{"DOI":"10.13039\/100010269","name":"Wellcome Trust","doi-asserted-by":"crossref","award":["217163\/Z\/19\/Z"],"award-info":[{"award-number":["217163\/Z\/19\/Z"]}],"id":[{"id":"10.13039\/100010269","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["mBio"],"published-print":{"date-parts":[[2024,4,10]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:sec>\n                    <jats:title\/>\n                    <jats:p>\n                      <jats:italic>Cryptococcus neoformans<\/jats:italic>\n                      causes lethal meningitis and accounts for approximately 10%\u201315% of AIDS-associated deaths worldwide. There are major gaps in our understanding of how this fungus invades the mammalian brain. To investigate the dynamics of\n                      <jats:italic>C. neoformans<\/jats:italic>\n                      tissue invasion, we mapped fungal localization and host cell interactions in infected brain, lung, and upper airways using mouse models of systemic and airway infection. To enable this, we developed an\n                      <jats:italic>in situ<\/jats:italic>\n                      imaging pipeline capable of measuring large volumes of tissue while preserving anatomical and cellular information by combining thick tissue sections, tissue clarification, and confocal imaging. We confirm high fungal burden in mouse upper airway after nasal inoculation. Yeast in turbinates were frequently titan cells, with faster kinetics than reported in mouse lungs. Importantly, we observed one instance of fungal cells enmeshed in lamina propria of the upper airways, suggesting penetration of airway mucosa as a possible route of tissue invasion and dissemination to the bloodstream. We extend previous literature positing bloodstream dissemination of\n                      <jats:italic>C. neoformans<\/jats:italic>\n                      , by finding viable fungi in the bloodstream of mice a few days after intranasal infection. As early as 24 h post systemic infection, the majority of\n                      <jats:italic>C. neoformans<\/jats:italic>\n                      cells traversed the blood-brain barrier, and were engulfed or in close proximity to microglia. Our work presents a new method for investigating microbial invasion, establishes that\n                      <jats:italic>C. neoformans<\/jats:italic>\n                      can breach multiple tissue barriers within the first days of infection, and demonstrates microglia as the first cells responding to\n                      <jats:italic>C. neoformans<\/jats:italic>\n                      invasion of the brain.\n                    <\/jats:p>\n                    <jats:sec>\n                      <jats:title>IMPORTANCE<\/jats:title>\n                      <jats:p>Cryptococcal meningitis causes 10%\u201315% of AIDS-associated deaths globally. Still, brain-specific immunity to cryptococci is a conundrum. By employing innovative imaging, this study reveals what occurs during the first days of infection in brain and in airways. We found that titan cells predominate in upper airways and that cryptococci breach the upper airway mucosa, which implies that, at least in mice, the upper airways are a site for fungal dissemination. This would signify that mucosal immunity of the upper airway needs to be better understood. Importantly, we also show that microglia, the brain-resident macrophages, are the first responders to infection, and microglia clusters are formed surrounding cryptococci. This study opens the field to detailed molecular investigations on airway immune response, how fungus traverses the blood-brain barrier, how microglia respond to infection, and ultimately how microglia monitor the blood-brain barrier to preserve brain function.<\/jats:p>\n                    <\/jats:sec>\n                  <\/jats:sec>","DOI":"10.1128\/mbio.03078-23","type":"journal-article","created":{"date-parts":[[2024,3,21]],"date-time":"2024-03-21T09:01:02Z","timestamp":1711011662000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":32,"title":["<i>Cryptococcus neoforman<\/i>\n                    s rapidly invades the murine brain by sequential breaching of airway and endothelial tissues barriers, followed by engulfment by microglia"],"prefix":"10.1128","volume":"15","author":[{"given":"Vanessa I.","family":"Francis","sequence":"first","affiliation":[{"name":"MRC Centre for Medical Mycology at University of Exeter, University of Exeter, Exeter, United Kingdom"},{"name":"Faculty of Health and Life Sciences, University of Exeter, Exeter, United Kingdom"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0001-8059-1730","authenticated-orcid":true,"given":"Corin","family":"Liddle","sequence":"additional","affiliation":[{"name":"Bioimaging Facility, University of Exeter, Exeter, United Kingdom"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Emma","family":"Camacho","sequence":"additional","affiliation":[{"name":"Johns Hopkins Bloomberg School of Public Health, Baltimore, Maryland, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Madhura","family":"Kulkarni","sequence":"additional","affiliation":[{"name":"Johns Hopkins Bloomberg School of Public Health, Baltimore, Maryland, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Samuel R. S.","family":"Junior","sequence":"additional","affiliation":[{"name":"Johns Hopkins Bloomberg School of Public Health, Baltimore, Maryland, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jamie A.","family":"Harvey","sequence":"additional","affiliation":[{"name":"MRC Centre for Medical Mycology at University of Exeter, University of Exeter, Exeter, United Kingdom"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8051-2302","authenticated-orcid":true,"given":"Elizabeth R.","family":"Ballou","sequence":"additional","affiliation":[{"name":"MRC Centre for Medical Mycology at University of Exeter, University of Exeter, Exeter, United Kingdom"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Darren D.","family":"Thomson","sequence":"additional","affiliation":[{"name":"MRC Centre for Medical Mycology at University of Exeter, University of Exeter, Exeter, United Kingdom"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0287-5383","authenticated-orcid":true,"given":"Gordon D.","family":"Brown","sequence":"additional","affiliation":[{"name":"MRC Centre for Medical Mycology at University of Exeter, University of Exeter, Exeter, United Kingdom"},{"name":"Faculty of Health and Life Sciences, University of Exeter, Exeter, United Kingdom"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4847-2045","authenticated-orcid":true,"given":"J. Marie","family":"Hardwick","sequence":"additional","affiliation":[{"name":"Johns Hopkins Bloomberg School of Public Health, Baltimore, Maryland, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9402-9167","authenticated-orcid":true,"given":"Arturo","family":"Casadevall","sequence":"additional","affiliation":[{"name":"Johns Hopkins Bloomberg School of Public Health, Baltimore, Maryland, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jonathan","family":"Witton","sequence":"additional","affiliation":[{"name":"Faculty of Health and Life Sciences, University of Exeter, Exeter, United Kingdom"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7523-3031","authenticated-orcid":true,"given":"Carolina","family":"Coelho","sequence":"additional","affiliation":[{"name":"MRC Centre for Medical Mycology at University of Exeter, University of Exeter, Exeter, United Kingdom"},{"name":"Faculty of Health and Life Sciences, University of Exeter, Exeter, United Kingdom"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_4_3_2","doi-asserted-by":"publisher","DOI":"10.1542\/peds.107.5.e66"},{"key":"e_1_3_4_4_2","doi-asserted-by":"publisher","DOI":"10.1016\/bs.adgen.2020.02.001"},{"key":"e_1_3_4_5_2","doi-asserted-by":"publisher","DOI":"10.1172\/JCI136223"},{"key":"e_1_3_4_6_2","doi-asserted-by":"publisher","DOI":"10.1590\/0001-3765201520140704"},{"key":"e_1_3_4_7_2","doi-asserted-by":"publisher","DOI":"10.1128\/mSphere.00483-19"},{"key":"e_1_3_4_8_2","doi-asserted-by":"publisher","DOI":"10.1172\/JCI41963"},{"key":"e_1_3_4_9_2","doi-asserted-by":"publisher","DOI":"10.1111\/cmi.12027"},{"key":"e_1_3_4_10_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.micinf.2015.08.017"},{"key":"e_1_3_4_11_2","doi-asserted-by":"publisher","DOI":"10.1002\/eji.201546239"},{"key":"e_1_3_4_12_2","doi-asserted-by":"publisher","DOI":"10.1128\/mBio.01101-14"},{"key":"e_1_3_4_13_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41551-021-00743-8"},{"key":"e_1_3_4_14_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-020-15329-2"},{"key":"e_1_3_4_15_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41593-018-0301-3"},{"key":"e_1_3_4_16_2","doi-asserted-by":"publisher","DOI":"10.1128\/mBio.02752-19"},{"key":"e_1_3_4_17_2","doi-asserted-by":"publisher","DOI":"10.1038\/s42003-022-04340-6"},{"key":"e_1_3_4_18_2","doi-asserted-by":"publisher","DOI":"10.1038\/srep18631"},{"key":"e_1_3_4_19_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1000953"},{"key":"e_1_3_4_20_2","doi-asserted-by":"publisher","DOI":"10.3389\/fcimb.2021.804059"},{"key":"e_1_3_4_21_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2020.01.028"},{"key":"e_1_3_4_22_2","doi-asserted-by":"publisher","DOI":"10.1097\/nen.0b013e3180517b46"},{"key":"e_1_3_4_23_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.1194637"},{"key":"e_1_3_4_24_2","doi-asserted-by":"publisher","DOI":"10.3389\/fnins.2019.01291"},{"key":"e_1_3_4_25_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2020.03.021"},{"key":"e_1_3_4_26_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1007777"},{"key":"e_1_3_4_27_2","doi-asserted-by":"publisher","DOI":"10.1128\/mBio.02183-16"},{"key":"e_1_3_4_28_2","doi-asserted-by":"publisher","DOI":"10.1111\/cmi.13330"},{"key":"e_1_3_4_29_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.micinf.2005.07.027"},{"key":"e_1_3_4_30_2","doi-asserted-by":"publisher","DOI":"10.1186\/1465-9921-9-9"},{"key":"e_1_3_4_31_2","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.01065-08"},{"key":"e_1_3_4_32_2","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.00334-09"},{"key":"e_1_3_4_33_2","doi-asserted-by":"publisher","DOI":"10.1111\/myc.12619"},{"key":"e_1_3_4_34_2","doi-asserted-by":"publisher","DOI":"10.1084\/jem.180.1.365"},{"key":"e_1_3_4_35_2","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.72.9.4985-4995.2004"},{"key":"e_1_3_4_36_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.ajpath.2018.03.015"},{"key":"e_1_3_4_37_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-023-43061-0"},{"key":"e_1_3_4_38_2","doi-asserted-by":"publisher","DOI":"10.1128\/mBio.01415-17"},{"key":"e_1_3_4_39_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.neuron.2022.10.020"},{"key":"e_1_3_4_40_2","doi-asserted-by":"publisher","DOI":"10.1111\/jnc.12477"},{"key":"e_1_3_4_41_2","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.06332-11"},{"key":"e_1_3_4_42_2","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.174.11.7226"},{"key":"e_1_3_4_43_2","doi-asserted-by":"publisher","DOI":"10.1242\/dmm.039123"},{"key":"e_1_3_4_44_2","doi-asserted-by":"publisher","DOI":"10.1007\/BF00437432"},{"key":"e_1_3_4_45_2","doi-asserted-by":"publisher","DOI":"10.1023\/a:1006809522931"},{"key":"e_1_3_4_46_2","doi-asserted-by":"publisher","DOI":"10.2460\/javma.236.10.1069"},{"key":"e_1_3_4_47_2","doi-asserted-by":"publisher","DOI":"10.1177\/20551169221074624"},{"key":"e_1_3_4_48_2","doi-asserted-by":"publisher","DOI":"10.1080\/13693780500036019"},{"key":"e_1_3_4_49_2","doi-asserted-by":"publisher","DOI":"10.3109\/13693786.2011.594967"},{"key":"e_1_3_4_50_2","doi-asserted-by":"publisher","DOI":"10.1080\/mmy.40.3.273.282"},{"key":"e_1_3_4_51_2","doi-asserted-by":"publisher","DOI":"10.3109\/13693786.2011.564216"},{"key":"e_1_3_4_52_2","first-page":"489","volume-title":"Cryptococcus","author":"Malik R","year":"2011","unstructured":"Malik R, Krockenberger MB, O\u2019Brien CR, Carter DA, Meyer W, Canfield PJ. 2011. Veterinary insights into cryptococcosis caused by Cryptococcus neoformans and Cryptococcus gattii, p 489\u2013504. In Cryptococcus"},{"key":"e_1_3_4_53_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.molimm.2015.09.008"},{"key":"e_1_3_4_54_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41577-023-00972-9"},{"key":"e_1_3_4_55_2","doi-asserted-by":"publisher","DOI":"10.1016\/0022-510x(94)90223-2"},{"key":"e_1_3_4_56_2","first-page":"1267","article-title":"Immunohistochemical localization of capsular polysaccharide antigen in the central nervous system cells in cryptococcal meningoencephalitis","volume":"148","author":"Lee SC","year":"1996","unstructured":"Lee SC, Casadevall A, Dickson DW. 1996. Immunohistochemical localization of capsular polysaccharide antigen in the central nervous system cells in cryptococcal meningoencephalitis. Am J Pathol 148:1267\u20131274.","journal-title":"Am J Pathol"},{"key":"e_1_3_4_57_2","doi-asserted-by":"publisher","DOI":"10.1016\/s0046-8177(96)90459-1"},{"key":"e_1_3_4_58_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pntd.0011068"},{"key":"e_1_3_4_59_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.chom.2022.08.017"},{"key":"e_1_3_4_60_2","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.00506-16"},{"key":"e_1_3_4_61_2","doi-asserted-by":"publisher","DOI":"10.1128\/spectrum.04315-22"},{"key":"e_1_3_4_62_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-019-12381-5"},{"key":"e_1_3_4_63_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1008361"},{"key":"e_1_3_4_64_2","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2016.00164"},{"key":"e_1_3_4_65_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M112.353375"},{"key":"e_1_3_4_66_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0068408"},{"key":"e_1_3_4_67_2","doi-asserted-by":"publisher","DOI":"10.1128\/mbio.02838-22"},{"key":"e_1_3_4_68_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0140-6736(00)49915-0"},{"key":"e_1_3_4_69_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0076-6879(10)70033-1"},{"key":"e_1_3_4_70_2","doi-asserted-by":"publisher","DOI":"10.1038\/nmeth.2888"},{"key":"e_1_3_4_71_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0122916"},{"key":"e_1_3_4_72_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.fgb.2019.103241"},{"key":"e_1_3_4_73_2","doi-asserted-by":"publisher","DOI":"10.1038\/nmeth.2019"},{"key":"e_1_3_4_74_2","doi-asserted-by":"publisher","DOI":"10.1007\/978-3-030-00934-2_30"},{"key":"e_1_3_4_75_2","doi-asserted-by":"publisher","DOI":"10.1111\/cmi.12240"},{"key":"e_1_3_4_76_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1469-7580.2005.00407.x"}],"container-title":["mBio"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/mbio.03078-23","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/mbio.03078-23","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,4,10]],"date-time":"2024-04-10T09:04:48Z","timestamp":1712739888000},"score":1,"resource":{"primary":{"URL":"https:\/\/journals.asm.org\/doi\/10.1128\/mbio.03078-23"}},"subtitle":[],"editor":[{"given":"Luis R.","family":"Martinez","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"editor"}]}],"short-title":[],"issued":{"date-parts":[[2024,4,10]]},"references-count":74,"journal-issue":{"issue":"4","published-print":{"date-parts":[[2024,4,10]]}},"alternative-id":["10.1128\/mbio.03078-23"],"URL":"https:\/\/doi.org\/10.1128\/mbio.03078-23","relation":{"has-preprint":[{"id-type":"doi","id":"10.1101\/2023.11.13.564824","asserted-by":"object"}]},"ISSN":["2150-7511"],"issn-type":[{"value":"2150-7511","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,4,10]]},"assertion":[{"value":"2023-11-14","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2024-02-29","order":1,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2024-03-21","order":2,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"e03078-23"}}