Extracellular Vesicles Derived From Talaromyces marneffei Yeasts Mediate Inflammatory Response in Macrophage Cells by Bioactive Protein Components

Abstract

Extracellular vesicles (EVs) loaded with proteins, nucleic acids, membrane lipids, and other virulence factors could participate in pathogenic processes in some fungi such as Cryptococcus neoformans and Candida albicans. However, the specific characteristics of EVs derived from Talaromyces marneffei (TM) still have not been figured out yet. In the present study, it has been observed that TM-derived EVs were a heterogeneous group of nanosized membrane vesicles (30–300 nm) under nanoparticle tracking analysis and transmission electron microscopy. The DiI-labeled EVs could be taken up by RAW 264.7 macrophage cells. Incubation of EVs with macrophages would result in increased expression levels of reactive oxygen species, nitric oxide, and some inflammatory factors including interleukin-1β, interleukin-6, interleukin-10, and tumor necrosis factor. Furthermore, the expression of co-stimulatory molecules (CD80, CD86, and MHC-II) was also increased in macrophages stimulated with EVs. The level of inflammatory factors secreted by macrophages showed a significant decrease when EVs were hydrolyzed by protease, while that of DNA and RNA hydrolase treatment remained unchanged. Subsequently, some virulence factors in EVs including heat shock protein, mannoprotein 1, and peroxidase were determined by liquid chromatography–tandem mass spectrometry. Taken together, our results indicated that the TM-derived EVs could mediate inflammatory response and its protein would play a key role in regulating the function of RAW 264.7 macrophage cells.

The RNA Content of Fungal Extracellular Vesicles: At the “Cutting-Edge” of Pathophysiology Regulation

“…In this analysis, the greatest abundance of exonic sRNAs was found in Pb18 (104), followed by Pl01 (27) and Pb3 (19). Moreover, unique exonic sRNAs were identified in Pb18 (89), Pb01 (21), and Pb3 (4). The assumption is that exonic sRNAs in EVs provide sRNA interference (ex-siRNA) and regulate the RNA interference (RNAi) machinery [18].…”

“…Currently, EVs have been isolated from many different cell types and fluids [ 9 ], as well as in a wide range of fungal species, including yeast species such as Histoplasma capsulatum , Sporothrix schenckii , Candida parapsilosis , Saccharomyces cerevisiae , Malassezia sympodialis , Paracoccidioides brasiliensis , Candida albicans , Pichia fermentans , Cryptococcus gattii , Sporothrix brasiliensis , Paracoccidioides lutzii , Candida glabrata , Candida tropicalis , Talaromyces marneffei , and Candida auris [ 10 , 11 , 12 , 13 , 14 , 15 , 16 , 17 , 18 , 19 , 20 , 21 , 22 ], along with a set of filamentous species, including Exophiala dermatitidis , Alternaria infectoria , Trichophyton interdigitale , Rhizopus delemar , Fusarium oxysporum f. sp. Vasinfectum , Trichoderma reesei , Aspergillus fumigatus , Aspergillus flavus , and Penicillium digitatum [ 23 , 24 , 25 , 26 , 27 , 28 , 29 , 30 ].…”

Cryptococcus extracellular vesicles properties and their use as vaccine platforms

“…EV proteinase K treatment was performed following the previously described protocol (Yang et al., 2021 ) with some modifications. Briefly, proteinase K was added to the EV suspension (0.17 μg of sterol) to a final concentration of 2 mg/ml in 0.02 μm‐pore filtered PBS.…”

Analysis of Cryptococcal Extracellular Vesicles: Experimental Approaches for Studying Their Diversity Among Multiple Isolates, Kinetics of Production, Methods of Separation, and Detection in Cultures of Titan Cells

“…Fungal extracellular vesicles (EVs) were first described in Cryptococcus neoformans in 2007 ( 1 ). Since then, the phenomenon of EV production has been studied in detail in C. neoformans and Cryptococcus gattii ( 2 – 7 ) and has been expanded to other species, including Histoplasma capsulatum ( 8 ), Saccharomyces cerevisiae ( 9 ), Malassezia sympodialis ( 10 ), Paracoccidioides brasiliensis ( 11 ), Candida albicans ( 12 ), Candida glabrata ( 13 ), Candida tropicalis , Candida parapsilosis ( 8 , 13 ), Candida auris ( 14 ), Trichophyton interdigitale ( 15 ), Sporothrix brasiliensis ( 16 ), Trichoderma reesei ( 17 ), Aspergillus fumigatus ( 18 ), Fusarium oxysporum ( 19 ), Zymoseptoria tritici ( 20 ), Alternaria infectoria ( 21 ), Penicillium digitatum ( 22 ), Pichia fermentans ( 23 ), and Talaromyces marneffei ( 24 ). In this group of 20 species, we estimate that EVs were detected in samples produced by more than 50 fungal isolates.…”