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Visualizing Non-lytic Exocytosis of Cryptococcus neoformans from Macrophages Using Digital Light Microscopy
Published on: October 21, 2014
Cryptococcus neoformans-Infected Macrophages Release Proinflammatory Extracellular Vesicles: Insight into Their
Lei Zhang1,2,3,4, Keming Zhang1,2, Hang Li1,2
1Department of Dermatology and Venereology, Changzheng Hospital, Second Military Medical University, Shanghai, China.
Abstract:
Cryptococcus neoformans causes deadly mycosis in immunocompromised individuals. Macrophages are key cells fighting against microbes. Extracellular vesicles (EVs) are cell-to-cell communication mediators. The roles of EVs from infected host cells in the interaction with Cryptococcus remain uninvestigated. Here, EVs from viable C. neoformans-infected macrophages reduced fungal burdens but led to shorter survival of infected mice. In vitro, EVs induced naive macrophages to an inflammatory phenotype. Transcriptome analysis showed that EVs from viable C. neoformans-infected macrophages activated immune-related pathways, including p53 in naive human and murine macrophages. Conserved analysis demonstrated that basic cell biological processes, including cell cycle and division, were activated by infection-derived EVs from both murine and human infected macrophages. Combined proteomics, lipidomics, and metabolomics of EVs from infected macrophages showed regulation of pathways such as extracellular matrix (ECM) receptors and phosphatidylcholine. This form of intermacrophage communication could serve to prepare cells at more distant sites of infection to resist C. neoformans infection.IMPORTANCECryptococcus neoformans causes cryptococcal meningitis, which is frequent in patients with HIV/AIDS, especially in less-developed countries. The incidence of cryptococcal meningitis is close to 1 million each year globally. Macrophages are key cells that protect the body against microbes, including C. neoformans Extracellular vesicles are a group of membrane structures that are released from cells such as macrophages that modulate cell activities via the transfer of materials such as proteins, lipids, and RNAs. In this study, we found that Cryptococcus neoformans-infected macrophages produce extracellular vesicles that enhance the inflammatory response in Cryptococcus-infected mice. These Cryptococcus neoformans-infected macrophage vesicles also showed higher fungicidal biological effects on inactivated macrophages. Using omics technology, unique protein and lipid signatures were identified in these extracellular vesicles. Transcriptome analysis showed that these vesicles activated immune-related pathways like p53 in naive macrophages. The understanding of this intermacrophage communication could provide potential targets for the design of therapeutic agents to fight this deadly mycosis.
Insights
Extracellular vesicles from Cryptococcus neoformans-infected macrophages reduce fungal load but shorten survival in mice. These vesicles prime naive macrophages for an inflammatory response, offering potential therapeutic targets for cryptomycosis.
Area of Science:
- Immunology
- Cell Biology
- Microbiology
Background:
- Macrophages are critical in combating Cryptococcus neoformans infections.
- Extracellular vesicles (EVs) mediate intercellular communication.
- The role of EVs from infected macrophages in Cryptococcus interaction is unexplored.
Purpose of the Study:
- Investigate the function of EVs derived from C. neoformans-infected macrophages.
- Determine the impact of these EVs on host immune response and fungal burden.
- Characterize the molecular content and pathways modulated by these EVs.
Main Methods:
- Isolation and characterization of EVs from infected macrophages.
- In vivo studies in mice to assess fungal burden and survival.
- In vitro experiments with naive macrophages.
- Transcriptome, proteomic, lipidomic, and metabolomic analyses of EVs.
Main Results:
- EVs from infected macrophages reduced fungal burdens in vivo but decreased host survival.
- EVs induced an inflammatory phenotype in naive macrophages.
- Transcriptome analysis revealed activation of immune pathways, including p53.
- Multi-omics data indicated regulation of ECM receptor and phosphatidylcholine pathways.
Conclusions:
- EVs from C. neoformans-infected macrophages represent a novel intermacrophage communication mechanism.
- This communication primes distant macrophages, potentially enhancing resistance to infection.
- Understanding these EVs may lead to new therapeutic strategies against cryptomycosis.

