Extracellular Vesicles from Pneumocystis carinii-infected rats impair fungal viability but are dispensable for macrophage functions

Pneumocystis spp. are host obligate fungal pathogens that can cause severe pneumonia in mammals and rely heavily on their host for essential nutrients. The lack of a sustainable in vitro culture system poses challenges in understanding their metabolism and the acquisition of essential nutrients from host lungs remains unexplored.
Transmission electron micrographs show Extracellular Vesicles (EVs) are found near Pneumocystisspp. within the lung. We hypothesized that EVs transport essential nutrients to the fungi during infection. To investigate this, EVs from P. carinii- and P. murina-infected rodents were biochemically and functionally characterized. These EVs contained host proteins involved in cellular, metabolic, and immune processes as well as proteins with homologs found in other fungal EV proteomes, indicating Pneumocystis may release EVs. Notably, EV uptake by P. carinii indicated their potential involvement in nutrient acquisition and indicated a possibility for using engineered..., EV purification
Rats (n = 12) and mice (n = 6) were sacrificed after 9 weeks and 6 weeks of infection, respectively. Age-matched, uninfected, and immunosuppressed rodents were used as control groups (n = 12 rats, 6 mice). BALF was collected by instillation of cold 0.22 μm-filtered phosphate buffered saline (PBS; 10 mL for rats and 1 mL x 3 for mice) into the bronchiolar and alveolar spaces and gently collected. EV collection and purification were performed in 3 independent experiments and each isolation was used as a technical replicate for experiments described below. Cellular debris was removed by centrifugation at 3400 x g for 15 minutes.
EVs were purified using previously described methods (Lobb et al., 2015). Briefly, BALF was filtered using 100kDa Amicon Ultra (Millipore, Darmstadt, Germany). The flowthrough was collected as EV-depleted samples. Size exclusion chromatography (SEC) was performed on filtered BALF samples using qEV10 columns and the Automatic Fraction Collector (Izon..., , GENERAL INFORMATION

1. Title of Dataset: Extracellular Vesicles from Pneumocystis carinii-Infected Rats Impair Fungal Viability but are Dispensable for Macrophage Functions

2. Author Information
A. Principal Investigator Contact Information
Name: Melanie Cushion
Institution: University of Cincinnati
Address: Cincinnati, OH USA
Email:

B. Associate or Co-investigator Contact Information
Name: Steven Sayson
Institution: University of Cincinnati
Address: Cincinnati, OH USA
Email:

3. Date of data collection (single date, range, approximate date): 2020-2023

SHARING/ACCESS INFORMATION

1. Licenses/restrictions placed on the data: CC0 1.0 Universal (CC0 1.0) Public Domain

2. Links to publications that cite or use the data:

Sayson, S. G., Ashbaugh, A., & Cushion, M. T. (2024). Extracellular Vesicles from Pneumocystis carinii-infected rats impair fungal viability but are dispensable for macrophage functions. Microbiology Spectrum.

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