Editorial

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Open AccessEditorial

Extracellular Vesicles in Microbes, Pathogens, and Infectious Diseases

by Franklin Wang-Ngai Chow and Russell M. Morphew

Int. J. Mol. Sci. 2023, 24(13), 10686; https://0-doi-org.brum.beds.ac.uk/10.3390/ijms241310686 - 26 Jun 2023

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Extracellular vesicles (EVs) are nanosized lipid bilayer particles that are produced by all kinds of organisms, including both pathogenic and non-pathogenic archaea, bacteria, fungi, and parasites [...] Full article

(This article belongs to the Special Issue Extracellular Vesicles in Microbes, Pathogens and Infectious Diseases)

Research

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18 pages, 2423 KiB

Open AccessArticle

The Metagenomic Composition and Effects of Fecal-Microbe-Derived Extracellular Vesicles on Intestinal Permeability Depend on the Patient’s Disease

by Cristina Rodríguez-Díaz, Flores Martín-Reyes, Bernard Taminiau, Ailec Ho-Plágaro, Raquel Camargo, Felix Fernandez-Garcia, José Pinazo-Bandera, Juan Pedro Toro-Ortiz, Montserrat Gonzalo, Carlos López-Gómez, Francisca Rodríguez-Pacheco, Dámaris Rodríguez de los Reyes, Georges Daube, Guillermo Alcain-Martinez and Eduardo García-Fuentes

Int. J. Mol. Sci. 2023, 24(5), 4971; https://0-doi-org.brum.beds.ac.uk/10.3390/ijms24054971 - 04 Mar 2023

Cited by 3 | Viewed by 1843

Abstract

The composition and impact of fecal-microbe-derived extracellular vesicles (EVs) present in different diseases has not been analyzed. We determined the metagenomic profiling of feces and fecal-microbe-derived EVs from healthy subjects and patients with different diseases (diarrhea, morbid obesity and Crohn’s disease (CD)) and [...] Read more.

The composition and impact of fecal-microbe-derived extracellular vesicles (EVs) present in different diseases has not been analyzed. We determined the metagenomic profiling of feces and fecal-microbe-derived EVs from healthy subjects and patients with different diseases (diarrhea, morbid obesity and Crohn’s disease (CD)) and the effect of these fecal EVs on the cellular permeability of Caco-2 cells. The control group presented higher proportions of Pseudomonas and Rikenellaceae_RC9_gut_group and lower proportions of Phascolarctobacterium, Veillonella and Veillonellaceae_ge in EVs when compared with the feces from which these EVs were isolated. In contrast, there were significant differences in 20 genera between the feces and EV compositions in the disease groups. Bacteroidales and Pseudomonas were increased, and Faecalibacterium, Ruminococcus, Clostridium and Subdoligranum were decreased in EVs from control patients compared with the other three groups of patients. Tyzzerella, Verrucomicrobiaceae, Candidatus_Paracaedibacter and Akkermansia were increased in EVs from the CD group compared with the morbid obesity and diarrhea groups. Fecal EVs from the morbid obesity, CD and, mainly, diarrhea induced a significant increase in the permeability of Caco-2 cells. In conclusion, the metagenomic composition of fecal-microbe-derived EVs changes depending on the disease of the patients. The modification of the permeability of Caco-2 cells produced by fecal EVs depends on the disease of the patients. Full article

(This article belongs to the Special Issue Extracellular Vesicles in Microbes, Pathogens and Infectious Diseases)

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17 pages, 3094 KiB

Open AccessArticle

Differential Effect of Extracellular Vesicles Derived from Plasmodium falciparum-Infected Red Blood Cells on Monocyte Polarization

by Ladawan Khowawisetsut, Sinmanus Vimonpatranon, Kittima Lekmanee, Hathai Sawasdipokin, Narinee Srimark, Kesinee Chotivanich and Kovit Pattanapanyasat

Int. J. Mol. Sci. 2023, 24(3), 2631; https://0-doi-org.brum.beds.ac.uk/10.3390/ijms24032631 - 30 Jan 2023

Cited by 4 | Viewed by 1746

Abstract

Malaria is a life-threatening tropical arthropod-borne disease caused by Plasmodium spp. Monocytes are the primary immune cells to eliminate malaria-infected red blood cells. Thus, the monocyte’s functions are one of the crucial factors in controlling parasite growth. It is reasoned that the activation [...] Read more.

Malaria is a life-threatening tropical arthropod-borne disease caused by Plasmodium spp. Monocytes are the primary immune cells to eliminate malaria-infected red blood cells. Thus, the monocyte’s functions are one of the crucial factors in controlling parasite growth. It is reasoned that the activation or modulation of monocyte function by parasite products might dictate the rate of disease progression. Extracellular vesicles (EVs), microvesicles, and exosomes, released from infected red blood cells, mediate intercellular communication and control the recipient cell function. This study aimed to investigate the physical characteristics of EVs derived from culture-adapted P. falciparum isolates (Pf-EVs) from different clinical malaria outcomes and their impact on monocyte polarization. The results showed that all P. falciparum strains released similar amounts of EVs with some variation in size characteristics. The effect of Pf-EV stimulation on M1/M2 monocyte polarization revealed a more pronounced effect on CD14⁺CD16⁺ intermediate monocytes than the CD14⁺CD16⁻ classical monocytes with a marked induction of Pf-EVs from a severe malaria strain. However, no difference in the levels of microRNAs (miR), miR-451a, miR-486, and miR-92a among Pf-EVs derived from virulent and nonvirulent strains was found, suggesting that miR in Pf-EVs might not be a significant factor in driving M2-like monocyte polarization. Future studies on other biomolecules in Pf-EVs derived from the P. falciparum strain with high virulence that induce M2-like polarization are therefore recommended. Full article

(This article belongs to the Special Issue Extracellular Vesicles in Microbes, Pathogens and Infectious Diseases)

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19 pages, 1434 KiB

Open AccessArticle

Characterization and Involvement of Exosomes Originating from Chikungunya Virus-Infected Epithelial Cells in the Transmission of Infectious Viral Elements

by Bao Chi Thi Le, Ati Burassakarn, Panwad Tongchai, Tipaya Ekalaksananan, Sirinart Aromseree, Supranee Phanthanawiboon, Yada Polsan, Neal Alexander, Hans J. Overgaard and Chamsai Pientong

Int. J. Mol. Sci. 2022, 23(20), 12117; https://0-doi-org.brum.beds.ac.uk/10.3390/ijms232012117 - 11 Oct 2022

Cited by 7 | Viewed by 1950

Abstract

The Chikungunya virus (CHIKV) is a mosquito-borne alphavirus that affects the world’s popula-tion with chikungunya disease. Adaptation of the viral life cycle to their host cells’ environment is a key step for establishing their infection and pathogenesis. Recently, the accumulating evidence advocates a [...] Read more.

The Chikungunya virus (CHIKV) is a mosquito-borne alphavirus that affects the world’s popula-tion with chikungunya disease. Adaptation of the viral life cycle to their host cells’ environment is a key step for establishing their infection and pathogenesis. Recently, the accumulating evidence advocates a principal role of extracellular vesicles (EVs), including exosomes, in both the infection and pathogenesis of infectious diseases. However, the participation of exosomes in CHIKV infec-tion and transmission is not well clarified. Here, we demonstrated that the CHIKV RNA and pro-teins were captured in exosomes, which were released by viral-infected epithelial cells. A viral genomic element in the isolated exosomes was infectious to naïve mammalian epithelial cells. The assay of particle size distribution and transmission electron microscopy (TEM) revealed CHIKV-derived exosomes with a size range from 50 to 250 nm. Treatments with RNase A, Triton X-100, and immunoglobulin G antibodies from CHIKV-positive patient plasma indicated that in-fectious viral elements are encompassed inside the exosomes. Interestingly, our viral plaque for-mation also exhibited that infectious viral elements might be securely transmitted to neighboring cells by a secreted exosomal pathway. Taken together, our recent findings emphasize the evidence for a complementary means of CHIKV infection and suggest the role of exosome-mediated CHIKV transmission. Full article

(This article belongs to the Special Issue Extracellular Vesicles in Microbes, Pathogens and Infectious Diseases)

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22 pages, 2290 KiB

Open AccessArticle

Fasciola hepatica Gastrodermal Cells Selectively Release Extracellular Vesicles via a Novel Atypical Secretory Mechanism

by Adam P. S. Bennett, Eduardo de la Torre-Escudero, Susan S. E. Dermott, Lawrence T. Threadgold, Robert E. B. Hanna and Mark W. Robinson

Int. J. Mol. Sci. 2022, 23(10), 5525; https://0-doi-org.brum.beds.ac.uk/10.3390/ijms23105525 - 15 May 2022

Cited by 6 | Viewed by 2681

Abstract

The liver fluke, Fasciola hepatica, is an obligate blood-feeder, and the gastrodermal cells of the parasite form the interface with the host’s blood. Despite their importance in the host–parasite interaction, in-depth proteomic analysis of the gastrodermal cells is lacking. Here, we used [...] Read more.

The liver fluke, Fasciola hepatica, is an obligate blood-feeder, and the gastrodermal cells of the parasite form the interface with the host’s blood. Despite their importance in the host–parasite interaction, in-depth proteomic analysis of the gastrodermal cells is lacking. Here, we used laser microdissection of F. hepatica tissue sections to generate unique and biologically exclusive tissue fractions of the gastrodermal cells and tegument for analysis by mass spectrometry. A total of 226 gastrodermal cell proteins were identified, with proteases that degrade haemoglobin being the most abundant. Other detected proteins included those such as proton pumps and anticoagulants which maintain a microenvironment that facilitates digestion. By comparing the gastrodermal cell proteome and the 102 proteins identified in the laser microdissected tegument with previously published tegument proteomic datasets, we showed that one-quarter of proteins (removed by freeze–thaw extraction) or one-third of proteins (removed by detergent extraction) previously identified as tegumental were instead derived from the gastrodermal cells. Comparative analysis of the laser microdissected gastrodermal cells, tegument, and F. hepatica secretome revealed that the gastrodermal cells are the principal source of secreted proteins, as well as showed that both the gastrodermal cells and the tegument are likely to release subpopulations of extracellular vesicles (EVs). Microscopical examination of the gut caeca from flukes fixed immediately after their removal from the host bile ducts showed that selected gastrodermal cells underwent a progressive thinning of the apical plasma membrane which ruptured to release secretory vesicles en masse into the gut lumen. Our findings suggest that gut-derived EVs are released via a novel atypical secretory route and highlight the importance of the gastrodermal cells in nutrient acquisition and possible immunomodulation by the parasite. Full article

(This article belongs to the Special Issue Extracellular Vesicles in Microbes, Pathogens and Infectious Diseases)

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13 pages, 2180 KiB

Open AccessArticle

Quality Control of Bacterial Extracellular Vesicles with Total Protein Content Assay, Nanoparticles Tracking Analysis, and Capillary Electrophoresis

by Aleksandra Steć, Joanna Jońca, Krzysztof Waleron, Małgorzata Waleron, Agata Płoska, Leszek Kalinowski, Bartosz Wielgomas and Szymon Dziomba

Int. J. Mol. Sci. 2022, 23(8), 4347; https://0-doi-org.brum.beds.ac.uk/10.3390/ijms23084347 - 14 Apr 2022

Cited by 10 | Viewed by 2918

Abstract

Extracellular vesicles (EVs) were isolated from Pectobacterium zantedeschiae culturing media using direct ultracentrifugation (UC), iodixanol cushion ultracentrifugation (ICUC), and iodixanol density gradient ultracentrifugation (IDGUC) techniques. The isolates were characterized with total protein content assay (bicinchoninic acid assay, BCA), nanoparticles tracking analysis (NTA), and [...] Read more.

Extracellular vesicles (EVs) were isolated from Pectobacterium zantedeschiae culturing media using direct ultracentrifugation (UC), iodixanol cushion ultracentrifugation (ICUC), and iodixanol density gradient ultracentrifugation (IDGUC) techniques. The isolates were characterized with total protein content assay (bicinchoninic acid assay, BCA), nanoparticles tracking analysis (NTA), and capillary electrophoresis (CE). A satisfactory correlation (R² > 0.94) between quantitative results obtained with BCA, NTA and CE was achieved only for isolates obtained with the IDGUC. The correlation between protein content and CE was proved to be related to the isolates’ purity. The NTA was found unable to provide reliable information on EVs quantity in samples isolated with UC and ICUC, due to the co-isolated particulate impurities. Moreover, the work reports polysaccharides, used as culturing media components, as a potential source of bias of quantitation with total protein content assay and NTA. The study demonstrates the advantageous selectivity of CE in quality control of EVs and its ability to differentiate subpopulations of EVs of Pectobacterium. Full article

(This article belongs to the Special Issue Extracellular Vesicles in Microbes, Pathogens and Infectious Diseases)

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18 pages, 2974 KiB

Open AccessArticle

Membrane Vesicles of Pectobacterium as an Effective Protein Secretion System

by Joanna Jonca, Malgorzata Waleron, Paulina Czaplewska, Aleksandra Bogucka, Aleksandra Steć, Szymon Dziomba, Jacek Jasiecki, Michał Rychłowski and Krzysztof Waleron

Int. J. Mol. Sci. 2021, 22(22), 12574; https://0-doi-org.brum.beds.ac.uk/10.3390/ijms222212574 - 22 Nov 2021

Cited by 9 | Viewed by 2215

Abstract

Bacteria of genus Pectobacterium are Gram-negative rods of the family Pectobacteriaceae. They are the causative agent of soft rot diseases of crops and ornamental plants. However, their virulence mechanisms are not yet fully elucidated. Membrane vesicles (MVs) are universally released by bacteria [...] Read more.

Bacteria of genus Pectobacterium are Gram-negative rods of the family Pectobacteriaceae. They are the causative agent of soft rot diseases of crops and ornamental plants. However, their virulence mechanisms are not yet fully elucidated. Membrane vesicles (MVs) are universally released by bacteria and are believed to play an important role in the pathogenicity and survival of bacteria in the environment. Our study investigates the role of MVs in the virulence of Pectobacterium. The results indicate that the morphology and MVs production depend on growth medium composition. In polygalacturonic acid (PGA) supplemented media, Pectobacterium produces large MVs (100–300 nm) and small vesicles below 100 nm. Proteomic analyses revealed the presence of pectate degrading enzymes in the MVs. The pectate plate test and enzymatic assay proved that those enzymes are active and able to degrade pectates. What is more, the pathogenicity test indicated that the MVs derived from Pectobacterium were able to induce maceration of Zantedeschia sp. leaves. We also show that the MVs of β-lactamase producing strains were able to suppress ampicillin activity and permit the growth of susceptible bacteria. Those findings indicate that the MVs of Pectobacterium play an important role in host-pathogen interactions and niche competition with other bacteria. Our research also sheds some light on the mechanism of MVs production. We demonstrate that the MVs production in Pectobacterium strains, which overexpress a green fluorescence protein (GFP), is higher than in wild-type strains. Moreover, proteomic analysis revealed that the GFP was present in the MVs. Therefore, it is possible that protein sequestration into MVs might not be strictly limited to periplasmic proteins. Our research highlights the importance of MVs production as a mechanism of cargo delivery in Pectobacterium and an effective secretion system. Full article

(This article belongs to the Special Issue Extracellular Vesicles in Microbes, Pathogens and Infectious Diseases)

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Review

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15 pages, 8146 KiB

Open AccessReview

Research Progress on Bacterial Membrane Vesicles and Antibiotic Resistance

by Xiaofei Liu, Jinyang Xiao, Shuming Wang, Jinxia Zhou, Jiale Qin, Zhibo Jia, Yanfeng Wang, Zhigang Wang, Yongmin Zhang and Huifang Hao

Int. J. Mol. Sci. 2022, 23(19), 11553; https://0-doi-org.brum.beds.ac.uk/10.3390/ijms231911553 - 30 Sep 2022

Cited by 10 | Viewed by 3158

Abstract

As a result of antibiotic overuse, bacterial antibiotic resistance has become a severe threat to worldwide public health. The development of more effective antimicrobial therapies and alternative antibiotic strategies is urgently required. The role played by bacterial membrane vesicles (BMVs) in antibiotic resistance [...] Read more.

As a result of antibiotic overuse, bacterial antibiotic resistance has become a severe threat to worldwide public health. The development of more effective antimicrobial therapies and alternative antibiotic strategies is urgently required. The role played by bacterial membrane vesicles (BMVs) in antibiotic resistance has become a current focus of research. BMVs are nanoparticles derived from the membrane components of Gram-negative and Gram-positive bacteria and contain diverse components originating from the cell envelope and cytoplasm. Antibiotic stress stimulates the secretion of BMVs. BMVs promote and mediate antibiotic resistance by multiple mechanisms. BMVs have been investigated as conceptually new antibiotics and drug-delivery vehicles. In this article, we outline the research related to BMVs and antibiotic resistance as a reference for the intentional use of BMVs to combat antibiotic resistance. Full article

(This article belongs to the Special Issue Extracellular Vesicles in Microbes, Pathogens and Infectious Diseases)

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