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Article: Comparison of Ultrastructure, Extracellular Matrix, and Drug Susceptibility in

McManus, William R / Schorey, Jeffrey S

2023 Volume 12, Issue 12

Abstract: Pulmonary infections ... ...

Abstract	Pulmonary infections with
Language	English
Publishing date	2023-12-08
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2695572-6
ISSN	2076-0817
ISSN	2076-0817
DOI	10.3390/pathogens12121427
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Article ; Online: Mycobacterium tuberculosis SecA2-dependent activation of host Rig-I/MAVs signaling is not conserved in Mycobacterium marinum.

Serene, Lindsay G / Webber, Kylie / Champion, Patricia A / Schorey, Jeffrey S

PloS one

2024 Volume 19, Issue 2, Page(s) e0281564

Abstract: Retinoic acid inducible gene I (Rig-I) is a cytosolic pattern recognition receptor canonically described for its important role in sensing viral RNAs. Increasingly, bacterially-derived RNA from intracellular bacteria such as Mycobacterium tuberculosis, ... ...

Abstract	Retinoic acid inducible gene I (Rig-I) is a cytosolic pattern recognition receptor canonically described for its important role in sensing viral RNAs. Increasingly, bacterially-derived RNA from intracellular bacteria such as Mycobacterium tuberculosis, have been shown to activate the same host Rig-I/Mitochondrial antiviral sensing protein (MAVS) signaling pathway to drive a type-I interferon response that contributes to bacterial pathogenesis in vivo. In M. tuberculosis, this response is mediated by the protein secretion system SecA2, but little is known about whether this process is conserved in other pathogenic mycobacteria or the mechanism by which these nucleic acids gain access to the host cytoplasm. Because the M. tuberculosis and M. marinum SecA2 protein secretion systems share a high degree of genetic and functional conservation, we hypothesized that Rig-I/MAVS activation and subsequent induction of IFN-β secretion by host macrophages will also be conserved between these two mycobacterial species. To test this, we generated a ΔsecA2 M. marinum strain along with complementation strains expressing either the M. marinum or M. tuberculosis secA2 genes. Our results suggest that the ΔsecA2 strain has a growth defect in vitro but not in host macrophages. These intracellular growth curves also suggested that the calculation applied to estimate the number of bacteria added to macrophage monolayers in infection assays underestimates bacterial inputs for the ΔsecA2 strain. Therefore, to better examine secreted IFN-β levels when bacterial infection levels are equal across strains we plated bacterial CFUs at 2hpi alongside our ELISA based infections. This enabled us to normalize secreted levels of IFN-β to a standard number of bacteria. Applying this approach to both WT and MAVS-/- bone marrow derived macrophages we observed equal or higher levels of secreted IFN-β from macrophages infected with the ΔsecA2 M. marinum strain as compared to WT. Together our findings suggest that activation of host Rig-I/MAVS cytosolic sensors and subsequent induction of IFN-β response in a SecA2-dependent manner is not conserved in M. marinum under the conditions tested.
MeSH term(s)	Humans ; Mycobacterium tuberculosis/genetics ; Mycobacterium marinum/genetics ; Signal Transduction ; Macrophages/metabolism ; DEAD Box Protein 58/metabolism ; Tuberculosis/pathology
Chemical Substances	DEAD Box Protein 58 (EC 3.6.4.13)
Language	English
Publishing date	2024-02-23
Publishing country	United States
Document type	Journal Article
ZDB-ID	2267670-3
ISSN	1932-6203 ; 1932-6203
ISSN (online)	1932-6203
ISSN	1932-6203
DOI	10.1371/journal.pone.0281564
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Article ; Online: Comparison of Ultrastructure, Extracellular Matrix, and Drug Susceptibility in M. avium subs. hominissuis Biofilms

William R. McManus / Jeffrey S. Schorey

Pathogens, Vol 12, Iss 12, p

2023 Volume 1427

Abstract: Pulmonary infections with Mycobacterium avium occur in susceptible individuals following exposure to the bacterium in the environment, where it often persists in biofilms. Many methods have been used to generate biofilms of M. avium , and it is unknown ... ...

Abstract	Pulmonary infections with Mycobacterium avium occur in susceptible individuals following exposure to the bacterium in the environment, where it often persists in biofilms. Many methods have been used to generate biofilms of M. avium , and it is unknown whether different approaches generate similar structures and cell phenotypes. To make a parallel comparison of in vitro biofilm ultrastructure, extracellular matrix (ECM) composition, and the drug susceptibility of biofilm resident bacteria, we used two published methods to generate M. avium biofilms: four-week incubation in M63 medium or 24 h exposure to dithiothreitol (DTT). Scanning electron microscopy revealed differences in the biofilm ultrastructure between the two methods, including variation in the appearance of ECM materials and morphology of resident cells, while light microscopy and staining with calcofluor white indicated that both biofilms contained polysaccharides characteristic of cellulose. Measuring the susceptibility of biofilms to degradation by enzymes suggested differences in structurally important ECM molecules, with DTT biofilms having important protein and, to a lesser extent, cellulose components, and M63 biofilms having moderate protein, cellulose, and DNA components. Both biofilms conferred resistance to the bactericidal effects of amikacin and clarithromycin, with resident cells being killed at greater than 10-fold lower rates than planktonic cells at almost all concentrations. These comparisons indicate differences in biofilm responses by M. avium under differing conditions, but also suggest common features of biofilm formation, including cellulose production and antimicrobial resistance.
Keywords	Mycobacterium avium ; biofilm ; resistance ; extracellular matrix ; Medicine ; R
Subject code	572
Language	English
Publishing date	2023-12-01T00:00:00Z
Publisher	MDPI AG
Document type	Article ; Online
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Article ; Online: Activation of host nucleic acid sensors by

Schorey, Jeffrey S / Vecchio, Joseph / McManus, William R / Ongalo, Joshua / Webber, Kylie

Critical reviews in microbiology

2023 Volume 50, Issue 2, Page(s) 224–240

Abstract: Although the importance of deoxyribonucleic acid (DNA) and ribonucleic acid (RNA) sensors in controlling viral infection is well established, their role in promoting an effective immune response to pathogens other than viruses is less clear. This is ... ...

Abstract	Although the importance of deoxyribonucleic acid (DNA) and ribonucleic acid (RNA) sensors in controlling viral infection is well established, their role in promoting an effective immune response to pathogens other than viruses is less clear. This is particularly true for infections with mycobacteria, as studies point to both protective and detrimental roles for activation of nucleic acid sensors in controlling a mycobacterial infection. Some of the contradiction likely stems from the use of different model systems and different mycobacterial species/strains as well as from which nucleic acid sensors were studied and what downstream effectors were evaluated. In this review, we will describe the different nucleic acid sensors that have been studied in the context of mycobacterial infections, and how the different studies compare. We conclude with a section on how nucleic acid sensor agonists have been used therapeutically and what further information is needed to enhance their potential as therapeutic agents.
MeSH term(s)	Humans ; Nucleic Acids ; Mycobacterium/genetics ; Mycobacterium Infections/microbiology
Chemical Substances	Nucleic Acids
Language	English
Publishing date	2023-12-28
Publishing country	England
Document type	Journal Article ; Review
ZDB-ID	1053620-6
ISSN	1549-7828 ; 1040-841X
ISSN (online)	1549-7828
ISSN	1040-841X
DOI	10.1080/1040841X.2023.2294904
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Article ; Online: Bacteria- and host-derived extracellular vesicles - two sides of the same coin?

Schorey, Jeffrey S / Cheng, Yong / McManus, William R

Journal of cell science

2021 Volume 134, Issue 11

Abstract: Intracellular bacterial pathogens spend portions of their life cycle both inside and outside host cells. While in these two distinct environments, they release or shed bacterial components, including virulence factors that promote their survival and ... ...

Abstract	Intracellular bacterial pathogens spend portions of their life cycle both inside and outside host cells. While in these two distinct environments, they release or shed bacterial components, including virulence factors that promote their survival and replication. Some of these components are released through extracellular vesicles, which are either derived from the bacteria themselves or from the host cells. Bacteria- and host-derived vesicles have been studied almost exclusively in isolation from each other, with little discussion of the other type of secreted vesicles, despite the fact that both are generated during an in vivo infection and both are likely play a role in bacterial pathogenesis and host immunity. In this Review, we aim to bridge this gap and discuss what we know of bacterial membrane vesicles in their generation and composition. We will compare and contrast this with the composition of host-derived vesicles with regard to bacterial components. We will also compare host cell responses to the different vesicles, with a focus on how these vesicles modulate the immune response, using Mycobacterium, Listeria and Salmonella as specific examples for these comparisons.
MeSH term(s)	Bacteria ; Extracellular Vesicles ; Secretory Vesicles ; Virulence Factors
Chemical Substances	Virulence Factors
Language	English
Publishing date	2021-06-03
Publishing country	England
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Review
ZDB-ID	2993-2
ISSN	1477-9137 ; 0021-9533
ISSN (online)	1477-9137
ISSN	0021-9533
DOI	10.1242/jcs.256628
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Article ; Online: Regulation and mechanisms of extracellular vesicle biogenesis and secretion.

D'Souza-Schorey, Crislyn / Schorey, Jeffrey S

Essays in biochemistry

2018 Volume 62, Issue 2, Page(s) 125–133

Abstract: EV (extracellular vesicle) biology is a rapidly expanding field. These heterogeneous membrane vesicles, which are shed from virtually all cell types, collectively represent a new dimension of intercellular communication in normal physiology and disease. ... ...

Abstract	EV (extracellular vesicle) biology is a rapidly expanding field. These heterogeneous membrane vesicles, which are shed from virtually all cell types, collectively represent a new dimension of intercellular communication in normal physiology and disease. They have been shown to deliver infectious and pathogenic agents to non-infected cells whereas in cancers they are thought to condition the tumor microenvironment. Their presence in body fluids and inherent capacity for systemic delivery point to their clinical promise. All of the above only intensifies the need to better understand the classification, mode of biogenesis, and contents of the different subtypes of EVs. This article focusses on vesicle subtypes labeled as exosomes and MVs (microvesicles) and discusses the biogenesis and release of these vesicles from cells.
MeSH term(s)	Biological Transport ; Body Fluids/metabolism ; Cell Communication ; Endocytosis ; Exosomes/metabolism ; Extracellular Vesicles/metabolism ; Extracellular Vesicles/physiology
Language	English
Publishing date	2018-05-15
Publishing country	England
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't ; Review
ISSN	1744-1358 ; 0071-1365
ISSN (online)	1744-1358
ISSN	0071-1365
DOI	10.1042/EBC20170078
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Article ; Online: Mycobacterium tuberculosis

Cheng, Yong / Schorey, Jeffrey S

The Journal of experimental medicine

2018 Volume 215, Issue 11, Page(s) 2919–2935

Abstract: RNA sensing pathways are key elements in a host immune response to viral pathogens, but little is known of their importance during bacterial infections. We found ... ...

Abstract	RNA sensing pathways are key elements in a host immune response to viral pathogens, but little is known of their importance during bacterial infections. We found that
MeSH term(s)	Adaptor Proteins, Signal Transducing/genetics ; Adaptor Proteins, Signal Transducing/immunology ; Animals ; Cytosol/immunology ; Cytosol/microbiology ; Cytosol/pathology ; DEAD Box Protein 58/genetics ; DEAD Box Protein 58/immunology ; DNA/genetics ; DNA/immunology ; Interferon Regulatory Factor-7/genetics ; Interferon Regulatory Factor-7/immunology ; Interferon-beta/genetics ; Interferon-beta/immunology ; Macrophages/immunology ; Macrophages/microbiology ; Macrophages/pathology ; Membrane Proteins/genetics ; Membrane Proteins/immunology ; Mice ; Mice, Knockout ; Mycobacterium tuberculosis/genetics ; Mycobacterium tuberculosis/immunology ; Mycobacterium tuberculosis/pathogenicity ; RNA/genetics ; RNA/immunology ; Signal Transduction/immunology ; Tuberculosis/genetics ; Tuberculosis/immunology ; Tuberculosis/pathology
Chemical Substances	Adaptor Proteins, Signal Transducing ; IPS-1 protein, mouse ; Interferon Regulatory Factor-7 ; Irf7 protein, mouse ; Membrane Proteins ; Sting1 protein, mouse ; RNA (63231-63-0) ; Interferon-beta (77238-31-4) ; DNA (9007-49-2) ; Ddx58 protein, mouse (EC 3.6.1.-) ; DEAD Box Protein 58 (EC 3.6.4.13)
Language	English
Publishing date	2018-10-18
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural
ZDB-ID	218343-2
ISSN	1540-9538 ; 0022-1007
ISSN (online)	1540-9538
ISSN	0022-1007
DOI	10.1084/jem.20180508
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Article ; Online: Innate Immune Responses to Tuberculosis.

Schorey, Jeffrey S / Schlesinger, Larry S

Microbiology spectrum

2017 Volume 4, Issue 6

Abstract: Tuberculosis remains one of the greatest threats to human health. The causative bacterium, Mycobacterium tuberculosis, is acquired by the respiratory route. It is exquisitely adapted to humans and is a prototypic intracellular pathogen of macrophages, ... ...

Abstract	Tuberculosis remains one of the greatest threats to human health. The causative bacterium, Mycobacterium tuberculosis, is acquired by the respiratory route. It is exquisitely adapted to humans and is a prototypic intracellular pathogen of macrophages, with alveolar macrophages being the primary conduit of infection and disease. However, M. tuberculosis bacilli interact with and are affected by several soluble and cellular components of the innate immune system which dictate the outcome of primary infection, most commonly a latently infected healthy human host, in whom the bacteria are held in check by the host immune response within the confines of tissue granuloma, the host histopathologic hallmark. Such individuals can develop active TB later in life with impairment in the immune system. In contrast, in a minority of infected individuals, the early host immune response fails to control bacterial growth, and progressive granulomatous disease develops, facilitating spread of the bacilli via infectious aerosols. The molecular details of the M. tuberculosis-host innate immune system interaction continue to be elucidated, particularly those occurring within the lung. However, it is clear that a number of complex processes are involved at the different stages of infection that may benefit either the bacterium or the host. In this article, we describe a contemporary view of the molecular events underlying the interaction between M. tuberculosis and a variety of cellular and soluble components and processes of the innate immune system.
MeSH term(s)	Animals ; Disease Models, Animal ; Host-Pathogen Interactions ; Humans ; Immunity, Innate ; Mycobacterium tuberculosis/immunology ; Tuberculosis/immunology
Language	English
Publishing date	2017-01-20
Publishing country	United States
Document type	Journal Article ; Review
ISSN	2165-0497
ISSN (online)	2165-0497
DOI	10.1128/microbiolspec.TBTB2-0010-2016
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Article ; Online: Extracellular vesicles and infectious diseases: new complexity to an old story.

Schorey, Jeffrey S / Harding, Clifford V

The Journal of clinical investigation

2016 Volume 126, Issue 4, Page(s) 1181–1189

Abstract: Exosomes and other extracellular microvesicles (ExMVs) have important functions in intercellular communication and regulation. During the course of infection, these vesicles can convey pathogen molecules that serve as antigens or agonists of innate ... ...

Abstract	Exosomes and other extracellular microvesicles (ExMVs) have important functions in intercellular communication and regulation. During the course of infection, these vesicles can convey pathogen molecules that serve as antigens or agonists of innate immune receptors to induce host defense and immunity, or that serve as regulators of host defense and mediators of immune evasion. These molecules may include proteins, nucleic acids, lipids, and carbohydrates. Pathogen molecules may be disseminated by incorporation into vesicles that are created and shed by host cells, or they may be incorporated into vesicles shed from microbial cells. Involvement of ExMVs in the induction of immunity and host defense is widespread among many pathogens, whereas their involvement in immune evasion mechanisms is prominent among pathogens that establish chronic infection and is found in some that cause acute infection. Because of their immunogenicity and enrichment of pathogen molecules, exosomes may also have potential in vaccine preparations and as diagnostic markers. Additionally, the ability of exosomes to deliver molecules to recipient cells raises the possibility of their use for drug/therapy delivery. Thus, ExMVs play a major role in the pathogenesis of infection and provide exciting potential for the development of novel diagnostic and therapeutic approaches.
MeSH term(s)	Animals ; Cell-Derived Microparticles/immunology ; Cell-Derived Microparticles/pathology ; Exosomes/immunology ; Exosomes/pathology ; Host-Pathogen Interactions ; Humans ; Immune Evasion ; Immunity, Innate ; Infection/immunology ; Infection/microbiology ; Infection/pathology
Keywords	covid19
Language	English
Publishing date	2016-04-01
Publishing country	United States
Document type	Journal Article ; Review
ZDB-ID	3067-3
ISSN	1558-8238 ; 0021-9738
ISSN (online)	1558-8238
ISSN	0021-9738
DOI	10.1172/JCI81132
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Article ; Online: Host cytosolic RNA sensing pathway promotes T Lymphocyte-mediated mycobacterial killing in macrophages.

Cheng, Yong / Kiene, Nicholas J / Tatarian, Alexandra / Eix, Emily F / Schorey, Jeffrey S

PLoS pathogens

2020 Volume 16, Issue 5, Page(s) e1008569

Abstract: Mycobacterial infection leads to activation of the RIG-I/MAVS/TBK1 RNA sensing pathway in macrophages but the consequences of this activation remains poorly defined. In this study, we determined that activation of this RNA sensing pathway stimulates ICAM- ...

Abstract	Mycobacterial infection leads to activation of the RIG-I/MAVS/TBK1 RNA sensing pathway in macrophages but the consequences of this activation remains poorly defined. In this study, we determined that activation of this RNA sensing pathway stimulates ICAM-1 expression in M.avium-infected macrophage through the inhibition of the E3 ubiquitin ligase CRL4COP1/DET1. CRL4 when active targets the transcription factor ETV5 for degradation by the ubiquitin-proteasome system. In the absence of the ETV5 transcription factor, ICAM-1 expression is significantly decreased. The M.avium-induced ICAM-1 production is required for the formation of immune synapse between infected macrophages and antigen-specific CD4+ T lymphocytes, and is essential for CD4+ T lymphocyte-mediated mycobacterial killing in vitro and in mice. This study demonstrates a previously undefined mechanism by which a host cytosolic RNA sensing pathway contributes to the interplay between mycobacteria infected macrophages and antigen-specific T lymphocytes.
MeSH term(s)	Adaptor Proteins, Signal Transducing/genetics ; Adaptor Proteins, Signal Transducing/immunology ; Animals ; CD4-Positive T-Lymphocytes/immunology ; CD4-Positive T-Lymphocytes/microbiology ; CD4-Positive T-Lymphocytes/pathology ; DEAD Box Protein 58/genetics ; DEAD Box Protein 58/immunology ; Macrophages/immunology ; Macrophages/microbiology ; Macrophages/pathology ; Mice ; Mice, Knockout ; Mycobacterium avium/immunology ; Protein Serine-Threonine Kinases/genetics ; Protein Serine-Threonine Kinases/immunology ; Tuberculosis/genetics ; Tuberculosis/immunology ; Tuberculosis/pathology
Chemical Substances	Adaptor Proteins, Signal Transducing ; IPS-1 protein, mouse ; Tbk1 protein, mouse (EC 2.7.1.-) ; Protein Serine-Threonine Kinases (EC 2.7.11.1) ; Ddx58 protein, mouse (EC 3.6.1.-) ; DEAD Box Protein 58 (EC 3.6.4.13)
Language	English
Publishing date	2020-05-28
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural
ZDB-ID	2205412-1
ISSN	1553-7374 ; 1553-7374
ISSN (online)	1553-7374
ISSN	1553-7374
DOI	10.1371/journal.ppat.1008569
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