Concentration-dependent protein loading of extracellular vesicles released by Histoplasma capsulatum after antibody treatment and its modulatory action upon macrophages
Abstract
Here, diverse pathogenic fungi secrete extracellular vesicles (EV) that contain macromolecules, including virulence factors that can modulate the host immune response. We recently demonstrated that the binding of monoclonal antibodies (mAb) modulates how Histoplasma capsulatum load and releases its extracellular vesicles (EV). In the present paper, we addressed a concentration-dependent impact on the fungus’ EV loading and release with different mAb, as well as the pathophysiological role of these EV during the host-pathogen interaction. We found that the mAbs differentially regulate EV content in concentration-dependent and independent manners. Enzymatic assays demonstrated that laccase activity in EV from H. capsulatum opsonized with 6B7 was reduced, but urease activity was not altered. The uptake of H. capsulatum by macrophages pre-treated with EV, presented an antibody concentration-dependent phenotype. The intracellular killing of yeast cells was potently inhibited in macrophages pre-treated with EV from 7B6 (non-protective) mAb-opsonized H. capsulatum and this inhibition was associated with a decrease in the reactive-oxygen species generated by these macrophages. In summary, our findings show that opsonization quantitatively and qualitatively modifies H. capsulatum EV load and secretion leading to distinct effects on the host’s immune effector mechanisms, supporting the hypothesis that EV sorting and secretion are dynamic mechanisms formore »
- Authors:
-
[4];
[3];
- Albert Einstein College of Medicine, Bronx, NY (United States); Univ. Federal de Minas Gerais, Minas Gerais (Brazil)
- Albert Einstein College of Medicine, Bronx, NY (United States)
- Pacific Northwest National Lab. (PNNL), Richland, WA (United States)
- National Univ. of Singapore (Singapore)
- Univ. Federal do Rio de Janeiro (UFRJ), Rio de Janeiro (Brazil)
- Publication Date:
- Research Org.:
- Pacific Northwest National Laboratory (PNNL), Richland, WA (United States)
- Sponsoring Org.:
- USDOE
- OSTI Identifier:
- 1441220
- Report Number(s):
- PNNL-SA-124828
Journal ID: ISSN 2045-2322; PII: 25665
- Grant/Contract Number:
- AC05-76RL01830
- Resource Type:
- Accepted Manuscript
- Journal Name:
- Scientific Reports
- Additional Journal Information:
- Journal Volume: 8; Journal Issue: 1; Journal ID: ISSN 2045-2322
- Publisher:
- Nature Publishing Group
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES
Citation Formats
Baltazar, Ludmila Matos, Zamith-Miranda, Daniel, Burnet, Meagan C., Choi, Hyungwon, Nimrichter, Leonardo, Nakayasu, Ernesto S., and Nosanchuk, Joshua D. Concentration-dependent protein loading of extracellular vesicles released by Histoplasma capsulatum after antibody treatment and its modulatory action upon macrophages. United States: N. p., 2018.
Web. doi:10.1038/s41598-018-25665-5.
Baltazar, Ludmila Matos, Zamith-Miranda, Daniel, Burnet, Meagan C., Choi, Hyungwon, Nimrichter, Leonardo, Nakayasu, Ernesto S., & Nosanchuk, Joshua D. Concentration-dependent protein loading of extracellular vesicles released by Histoplasma capsulatum after antibody treatment and its modulatory action upon macrophages. United States. https://doi.org/10.1038/s41598-018-25665-5
Baltazar, Ludmila Matos, Zamith-Miranda, Daniel, Burnet, Meagan C., Choi, Hyungwon, Nimrichter, Leonardo, Nakayasu, Ernesto S., and Nosanchuk, Joshua D. Wed .
"Concentration-dependent protein loading of extracellular vesicles released by Histoplasma capsulatum after antibody treatment and its modulatory action upon macrophages". United States. https://doi.org/10.1038/s41598-018-25665-5. https://www.osti.gov/servlets/purl/1441220.
@article{osti_1441220,
title = {Concentration-dependent protein loading of extracellular vesicles released by Histoplasma capsulatum after antibody treatment and its modulatory action upon macrophages},
author = {Baltazar, Ludmila Matos and Zamith-Miranda, Daniel and Burnet, Meagan C. and Choi, Hyungwon and Nimrichter, Leonardo and Nakayasu, Ernesto S. and Nosanchuk, Joshua D.},
abstractNote = {Here, diverse pathogenic fungi secrete extracellular vesicles (EV) that contain macromolecules, including virulence factors that can modulate the host immune response. We recently demonstrated that the binding of monoclonal antibodies (mAb) modulates how Histoplasma capsulatum load and releases its extracellular vesicles (EV). In the present paper, we addressed a concentration-dependent impact on the fungus’ EV loading and release with different mAb, as well as the pathophysiological role of these EV during the host-pathogen interaction. We found that the mAbs differentially regulate EV content in concentration-dependent and independent manners. Enzymatic assays demonstrated that laccase activity in EV from H. capsulatum opsonized with 6B7 was reduced, but urease activity was not altered. The uptake of H. capsulatum by macrophages pre-treated with EV, presented an antibody concentration-dependent phenotype. The intracellular killing of yeast cells was potently inhibited in macrophages pre-treated with EV from 7B6 (non-protective) mAb-opsonized H. capsulatum and this inhibition was associated with a decrease in the reactive-oxygen species generated by these macrophages. In summary, our findings show that opsonization quantitatively and qualitatively modifies H. capsulatum EV load and secretion leading to distinct effects on the host’s immune effector mechanisms, supporting the hypothesis that EV sorting and secretion are dynamic mechanisms for a fine-tuned response by fungal cells.},
doi = {10.1038/s41598-018-25665-5},
journal = {Scientific Reports},
number = 1,
volume = 8,
place = {United States},
year = {Wed May 23 00:00:00 EDT 2018},
month = {Wed May 23 00:00:00 EDT 2018}
}
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Figures / Tables:
Figure 1: Protein and sterol quantification in EV from H. capsulatum. Yeast cells were incubated with or without 6 and 20 μg/mL of 6B7 and 7B6 mAb. The EV protein content was determined by BCA assay (A). EV sterol quantification was performed using Amplex reagent kit (B). Graphs represent meansmore »
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Bacillus anthracis produces membrane-derived vesicles containing biologically active toxins
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Antibody Therapy for Histoplasmosis
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- Frontiers in Microbiology, Vol. 7
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Works referencing / citing this record:
A Novel Protocol for the Isolation of Fungal Extracellular Vesicles Reveals the Participation of a Putative Scramblase in Polysaccharide Export and Capsule Construction in Cryptococcus gattii
journal, March 2019
- Reis, Flavia C. G.; Borges, Beatriz S.; Jozefowicz, Luísa J.
- mSphere, Vol. 4, Issue 2
Extracellular Vesicle-Mediated RNA Release in Histoplasma capsulatum
journal, March 2019
- Alves, Lysangela R.; Peres da Silva, Roberta; Sanchez, David A.
- mSphere, Vol. 4, Issue 2
Fungal Extracellular Vesicles as Potential Targets for Immune Interventions
journal, October 2019
- Freitas, Mateus Silveira; Bonato, Vânia Luiza Deperon; Pessoni, Andre Moreira
- mSphere, Vol. 4, Issue 6
A Novel Protocol for the Isolation of Fungal Extracellular Vesicles Reveals the Participation of a Putative Scramblase in Polysaccharide Export and Capsule Construction in Cryptococcus gattii
journal, March 2019
- Reis, Flavia C. G.; Borges, Beatriz S.; Jozefowicz, Luísa J.
- mSphere, Vol. 4, Issue 2
Extracellular Vesicle-Mediated RNA Release in Histoplasma capsulatum
journal, March 2019
- Alves, Lysangela R.; Peres da Silva, Roberta; Sanchez, David A.
- mSphere, Vol. 4, Issue 2
Fungal Extracellular Vesicles as Potential Targets for Immune Interventions
journal, October 2019
- Freitas, Mateus Silveira; Bonato, Vânia Luiza Deperon; Pessoni, Andre Moreira
- mSphere, Vol. 4, Issue 6
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