{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,1,16]],"date-time":"2026-01-16T00:25:14Z","timestamp":1768523114587,"version":"3.49.0"},"posted":{"date-parts":[[2025,7,18]]},"group-title":"Microbiology","reference-count":90,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2025,7,18]],"date-time":"2025-07-18T00:00:00Z","timestamp":1752796800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nd\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2025,7,18]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>\n                  <jats:italic>Cryptococcus neoformans,<\/jats:italic>\n                  the etiological agent of cryptococcal meningitis (CM), is a globally distributed environmental yeast that mainly causes infections in immunocompromised individuals. Particularly in low-resource countries, the mortality rate of CM can reach 81% and accounts for 19% of HIV\/AIDS-related deaths each year. In immunocompromised individuals, once inhaled,\n                  <jats:italic>C. neoformans<\/jats:italic>\n                  escapes from the lungs and disseminates with special predilection for the central nervous system (CNS). Once in the brain,\n                  <jats:italic>C. neoformans<\/jats:italic>\n                  interacts with microglia, the tissue-resident macrophages of the CNS. Previous studies indirectly showed that microglia are ineffective at controlling this fungal infection. The mechanisms underlying this fungal survival and proliferation within the CNS, however, remain unclear. In this study, we use and validate the C20 immortalized human microglia cell line to study cryptococcal-microglia interactions. We show that microglia have limited phagocytic activity that is specific to\n                  <jats:italic>C. neoformans<\/jats:italic>\n                  and partly dependent on cryptococcal antiphagocytic proteins that alter cell size and cell wall structure. We also show human microglia respond to cryptococcal strains differently than peripheral macrophages. Further, we show that human microglia are ineffective at killing phagocytosed\n                  <jats:italic>C. neoformans<\/jats:italic>\n                  , and that this could be due to the ability of this yeast to disrupt phagosome maturation and induce phagosome membrane damage in these cells. These findings provide us with fundamental knowledge regarding cryptococcal pathogenesis in the CNS, specifically insight into how\n                  <jats:italic>C. neoformans<\/jats:italic>\n                  is recognized by microglia under different conditions and demonstrate the usefulness of C20 cells to further study how this yeast survives and replicates within the CNS environment.\n                <\/jats:p>\n                <jats:sec>\n                  <jats:title>Importance<\/jats:title>\n                  <jats:p>\n                    While\n                    <jats:italic>Cryptococcus neoformans<\/jats:italic>\n                    is acquired through inhalation, the fatal pathology of cryptococcal infection occurs when the yeast disseminates to the central nervous system (CNS) and causes cryptococcal meningitis. Microglia are the first immune cells that\n                    <jats:italic>C. neoformans<\/jats:italic>\n                    will encounter once it reaches the CNS, and they are the largest population of macrophages in the brain. While microglia are professional phagocytes, they are unable to control\n                    <jats:italic>C. neoformans<\/jats:italic>\n                    infection. The mechanisms behind uncontrolled growth of\n                    <jats:italic>C. neoformans<\/jats:italic>\n                    within the CNS remains understudied, partly due to a lack of knowledge regarding microglia-cryptococcal interactions. This study provides fundamental knowledge into these interactions and establishes a powerful model to specifically study how\n                    <jats:italic>C. neoformans<\/jats:italic>\n                    is recognized by microglia and how cryptococcal phagosomes mature in these phagocytes. This work opens new avenues of research to further our understanding of cryptococcal-host interactions which can be leveraged to develop more effective therapeutics for cryptococcal meningitis.\n                  <\/jats:p>\n                <\/jats:sec>","DOI":"10.1101\/2025.07.17.665466","type":"posted-content","created":{"date-parts":[[2025,7,18]],"date-time":"2025-07-18T17:10:21Z","timestamp":1752858621000},"source":"Crossref","is-referenced-by-count":0,"title":["Unique recognition, phagocytosis, and intracellular survival of\n                  <i>Cryptococcus neoformans<\/i>\n                  in a human immortalized microglia cell line"],"prefix":"10.64898","author":[{"given":"Robbi L.","family":"Ross","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kassandra","family":"Arias-Parbul","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Zane M.","family":"Douglass","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Katrina L.","family":"Adams","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6453-4904","authenticated-orcid":false,"given":"Felipe H.","family":"Santiago-Tirado","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"54368","reference":[{"key":"2025072116300898000_2025.07.17.665466v1.1","doi-asserted-by":"crossref","unstructured":"Hawksworth DL , Lucking R . 2017. 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