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In Vivo Characterization of Candida Extracellular Vesicles Reveals Unique Infection Pathway Proteins and Validates In Vitro Observations

ABSTRACT: Extracellular vesicles (EVs) are membrane-bound particles secreted by cells, including microbes, and play key roles in intercellular communication and infection outcomes. Due to similarities between microbial and host EVs, isolating pathogen-specific EVs in vivo has been challenging. This study developed an immunoaffinity method using polyclonal antibodies against Candida albicans biofilm-derived EVs to selectively isolate fungal EVs from a rat catheter infection model. The method enabled specific capture and release of intact fungal EVs for analysis, without detecting host EVs. Characterization revealed EVs consistent with exosomes (~100 nm), and confirmed reduced EV production in a vesicle-defective Candida mutant (Hse1ΔΔ), validating in vitro findings. Proteomic analysis uncovered both previously known and novel Candida proteins, including the kinase CBK1, suggesting EV cargo adapts to in vivo conditions. This approach supports targeted EV study in infectious disease models and may aid in diagnostics and pathogenesis research.

INSTRUMENT(S):

ORGANISM(S): Candida Albicans Sc5314

TISSUE(S): Primary Cell, Extracellular Exosome

SUBMITTER: Greg Sabat

LAB HEAD: Justin Massey

PROVIDER: PXD067392 | Pride | 2026-04-06

REPOSITORIES: Pride

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			Action	DRS
	1-Serum-input_Andes_072324_21.raw	Raw
	1-Serum-input_Andes_072324_21__F004954_.mzid	Mzid
	1-Serum-input_Andes_072324_21__F004954_.mzid_1-Serum-input_Andes_072324_21__F004954_.MGF	Mzid
	10-IP_Andes_071824_17.raw	Raw
	10-IP_Andes_071824_17__F004963_.mzid	Mzid

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Project description:The majority of transplanted organs are recovered from deceased donors after brain death (BD). BD has been hypothesized to compromise organ quality in part from the activation of systemic inflammation. The objective of this study was to characterize the immune response induced by BD in a well controlled non-human primate (NHP) model. Assessment of physiologic parameters (blood pressure, heart rate, urinary output, catecholamines, and cerebral angiograms) was used to confirm BD. After 6h of BD, we monitored changes in the peripheral blood by flow cytometry, liver gene expression by microarray and liver protein expression by Western blotting and immunohistochemistry (IHC). BD was indicated by a rapid increase in blood pressure followed by hemodynamic instability, hypotension, diabetes insipidus and the absence of cerebral blood flow and brain stem reflexes. Within the peripheral blood IL-6 levels and neutrophils increased and myeloid dendritic cells decreased in BD NHP when compared to living donor controls. Genes related to innate inflammatory response and apoptosis were significantly upregulated in BD NHP. BD livers showed increased expression of suppressor of cytokine signaling 3 (SOCS3) protein and the danger associated molecular pattern protein S100A9. Increased expression of intracellular cellular adhesion molecule 1 (ICAM-1) and major histocompatibility complex (MHC) II, neutrophil accumulation, and products of oxidative stress (carboxy methyl lysine (CML) and hydroxynonenal (HNE)) were detected by IHC in livers. Conclusion: These data indicate that BD leads to the rapid activation of an inflammatory response within the liver involving components of the innate immune response at the gene and protein levels. The activation of these inflammatory pathways may provide one explanation for the reduced post-transplant function of organs from brain dead donors. Microarray analysis was performed to compare gene expression patterns from livers recovered after 5 hours of brain death (n=5) and from "living donor" non-brain dead controls (n=5).

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In Vivo Characterization of Candida Extracellular Vesicles Reveals Unique Infection Pathway Proteins and Validates In Vitro Observations

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