Related Experiment Video
Updated: Jun 23, 2026
Visualizing Non-lytic Exocytosis of Cryptococcus neoformans from Macrophages Using Digital Light Microscopy
Published on: October 21, 2014
Quantitative analysis reveals internalisation of Cryptococcus neoformans by brain endothelial cells in vivo
Yanli Chen1, Chang Li1, Donglei Sun1
1Division of Immunology, Virginia-Maryland College of Veterinary Medicine and Maryland Pathogen Research Institute, University of Maryland, College Park, Maryland, USA.
Abstract:
Migration of Cryptococcus neoformans from the blood to the brain parenchyma is crucial to cause fatal meningoencephalitis. Although mechanisms involved in brain migration of C. neoformans have been widely studied in vitro, less is known about how the fungus crosses the blood-brain barrier (BBB) in vivo. This is in part because of the lack of an approach to quantitatively analyse the dynamics of fungal transmigration into the brain across the BBB in vivo. In this study, we report a novel approach to quantitatively analyse the interactions between C. neoformans and brain endothelial cells in a mouse model using flow cytometry. Using this system, we show that C. neoformans was internalised by brain endothelial cells in vivo and that mice infected with acapsular or heat-killed C. neoformans yeast cells displayed a lower frequency of brain endothelial cells containing the yeast cell compared to mice infected with wild-type or viable yeast cells, respectively. We further demonstrate that brain endothelial cells were invaded by serotype A strain (H99 strain) at a higher rate compared to serotype D strain (52D strain). Our experiments established that internalisation of C. neoformans by brain endothelial cells occurred in vivo and offered a powerful approach to quantitatively analyse fungal migration into the brain.
Insights
Cryptococcus neoformans invades brain endothelial cells in vivo, a crucial step in meningoencephalitis. A new flow cytometry method quantifies this fungal brain migration, revealing strain and cell viability impacts.
Area of Science:
- Mycology
- Immunology
- Neuroscience
Background:
- Cryptococcus neoformans causes fatal meningoencephalitis by migrating from blood to the brain.
- Understanding fungal crossing of the blood-brain barrier (BBB) in vivo is limited.
- Quantitative analysis of fungal transmigration across the BBB in vivo is lacking.
Purpose of the Study:
- To develop and utilize a novel quantitative approach to analyze Cryptococcus neoformans interactions with brain endothelial cells in vivo.
- To investigate the in vivo dynamics of fungal transmigration across the blood-brain barrier.
Main Methods:
- Developed a novel flow cytometry-based approach in a mouse model.
- Quantitatively analyzed interactions between C. neoformans and brain endothelial cells in vivo.
- Compared infection rates using different fungal strains (serotype A vs. D) and conditions (viable vs. heat-killed, capsular vs. acapsular).
Main Results:
- Confirmed in vivo internalization of C. neoformans by brain endothelial cells.
- Demonstrated that viable and encapsulated wild-type yeast cells were internalized more frequently than heat-killed or acapsular cells.
- Showed higher invasion rates by serotype A (H99) compared to serotype D (52D) C. neoformans strains.
Conclusions:
- Established that brain endothelial cells internalize C. neoformans in vivo.
- The developed flow cytometry method provides a powerful tool for quantitative analysis of fungal brain migration.
- Findings offer insights into C. neoformans pathogenesis and BBB interactions.
More Related Videos
11:07Macrophage Cholesterol Depletion and Its Effect on the Phagocytosis of Cryptococcus neoformans
Published on: December 19, 2014
10:04Quantification of Cerebral Vascular Architecture using Two-photon Microscopy in a Mouse Model of HIV-induced Neuroinflammation
Published on: January 12, 2016
