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      Cryptococcus gattii alters immunostimulatory potential in response to the environment

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          Abstract

          Cryptococcus gattii is a capsular fungal pathogen, which causes life-threatening cryptococcosis in immunocompetent individuals. This emerging pathogen is less likely to be recognized by innate immunity compared to traditional Cryptococcus neoformans strains. Previous studies indicate that C-type lectin receptors (CLRs), including dectin-1 and dectin-2, play a role in recognizing cryptococcal cells; however, it remains to be elucidated whether the receptors physically associate with C. gattii yeast cell surfaces. Based on the previous findings, we hypothesized that culture conditions influence the expression or exposure of CLR ligands on C. gattii. Therefore, in the present study, we first investigated the culture conditions that induce exposure of CLR ligands on C. gattii yeast cells via the binding assay using recombinant fusion proteins of mouse CLR and IgG Fc, Fc dectin-1 and Fc dectin-2. Common fungal culture media, such as yeast extract–peptone–dextrose (YPD) broth, Sabouraud broth, and potato dextrose agar, did not induce the exposure of dectin-1 ligands, including β-1,3-glucan, on both capsular and acapsular C. gattii strains, in contrast to Fc dectin-1 and Fc dectin-2 bound to C. gattii cells growing in the conventional synthetic dextrose (SD) medium [may also be referred to as a yeast nitrogen base with glucose medium]. The medium also induced the exposure of dectin-1 ligands on C. neoformans, whereas all tested media induced dectin-1 and dectin-2 ligands in a control fungus Candida albicans. Notably, C. gattii did not expose dectin-1 ligands in SD medium supplemented with yeast extract or neutral buffer. In addition, compared to YPD medium-induced C. gattii, SD medium-induced C. gattii more efficiently induced the phosphorylation of Syk, Akt, and Erk1/2 in murine dendritic cells (DCs). Afterwards, the cells were considerably engulfed by DCs and remarkably induced DCs to secrete the inflammatory cytokines. Overall, the findings suggest that C. gattii alters its immunostimulatory potential in response to the environment.

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          Most cited references49

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          Immune recognition. A new receptor for beta-glucans.

          The carbohydrate polymers known as beta-1,3-d-glucans exert potent effects on the immune system - stimulating antitumour and antimicrobial activity, for example - by binding to receptors on macrophages and other white blood cells and activating them. Although beta-glucans are known to bind to receptors, such as complement receptor 3 (ref. 1), there is evidence that another beta-glucan receptor is present on macrophages. Here we identify this unknown receptor as dectin-1 (ref. 2), a finding that provides new insights into the innate immune recognition of beta-glucans.
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            C-type lectins in immunity and homeostasis

            The C-type lectins are a superfamily of proteins that recognize a broad repertoire of ligands and that regulate a diverse range of physiological functions. Most research attention has focused on the ability of C-type lectins to function in innate and adaptive antimicrobial immune responses, but these proteins are increasingly being recognized to have a major role in autoimmune diseases and to contribute to many other aspects of multicellular existence. Defects in these molecules lead to developmental and physiological abnormalities, as well as altered susceptibility to infectious and non-infectious diseases. In this Review, we present an overview of the roles of C-type lectins in immunity and homeostasis, with an emphasis on the most exciting recent discoveries.
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              Dectin-1 mediates macrophage recognition of Candida albicans yeast but not filaments.

              The ability of Candida albicans to rapidly and reversibly switch between yeast and filamentous morphologies is crucial to pathogenicity, and it is thought that the filamentous morphology provides some advantage during interaction with the mammalian immune system. Dectin-1 is a receptor that binds beta-glucans and is important for macrophage phagocytosis of fungi. The receptor also collaborates with Toll-like receptors for inflammatory activation of phagocytes by fungi. We show that yeast cell wall beta-glucan is largely shielded from Dectin-1 by outer wall components. However, the normal mechanisms of yeast budding and cell separation create permanent scars which expose sufficient beta-glucan to trigger antimicrobial responses through Dectin-1, including phagocytosis and activation of reactive oxygen production. During filamentous growth, no cell separation or subsequent beta-glucan exposure occurs, and the pathogen fails to activate Dectin-1. The data demonstrate a mechanism by which C. albicans shape alone directly contributes to the method by which phagocytes recognize the fungus.
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                Author and article information

                Contributors
                Role: ConceptualizationRole: Data curationRole: Formal analysisRole: Funding acquisitionRole: InvestigationRole: MethodologyRole: Project administrationRole: SupervisionRole: ValidationRole: VisualizationRole: Writing – original draftRole: Writing – review & editing
                Role: Data curationRole: Formal analysisRole: InvestigationRole: ValidationRole: Writing – review & editing
                Role: Data curationRole: Formal analysisRole: InvestigationRole: ValidationRole: Writing – review & editing
                Role: Data curationRole: InvestigationRole: ValidationRole: Writing – review & editing
                Role: ResourcesRole: Writing – review & editing
                Role: ResourcesRole: Writing – review & editing
                Role: Funding acquisitionRole: ResourcesRole: Writing – review & editing
                Role: Editor
                Journal
                PLoS One
                PLoS ONE
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, CA USA )
                1932-6203
                9 August 2019
                2019
                : 14
                : 8
                : e0220989
                Affiliations
                [1 ] Department of Chemotherapy and Mycoses, National Institute of Infectious Diseases, Toyama, Shinjuku-ku, Tokyo, Japan
                [2 ] Department of Biological Science and Technology, Faculty of Industrial Science and Technology, Tokyo University of Science, Niijuku, Katsushika-ku, Tokyo, Japan
                Wadsworth Center, UNITED STATES
                Author notes

                Competing Interests: The authors declare that no financial support, including that from Tomy Digital Biology Co. Ltd, alters their adherence to PLOS ONE policies on sharing data and materials.

                Author information
                http://orcid.org/0000-0003-0671-0049
                Article
                PONE-D-19-13717
                10.1371/journal.pone.0220989
                6688814
                31398236
                d6ef586b-7a20-48e0-82d2-318690bb6b80
                © 2019 Ueno et al

                This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

                History
                : 15 May 2019
                : 28 July 2019
                Page count
                Figures: 5, Tables: 0, Pages: 18
                Funding
                This work was supported by: Ministry of Education, Culture, Sports, Science, and Technology of Japan [KAKENHI 17K18385], Takeda Science Foundation, Tomy Digital Biology Co. Ltd [LEGEND Research Grant Program 2015] to Keigo Ueno; and by Ministry of Education, Culture, Sports, Science, and Technology of Japan [KAKENHI 17K10040], Japan Agency for Medical Research and Development, AMED [JP19fk0108045 and JP19fk0108094] to Yoshitsugu Miyazaki. These funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
                Categories
                Research Article
                Biology and Life Sciences
                Organisms
                Eukaryota
                Fungi
                Cryptococcus
                Cryptococcus Gattii
                Biology and Life Sciences
                Microbiology
                Medical Microbiology
                Microbial Pathogens
                Fungal Pathogens
                Cryptococcus Gattii
                Medicine and Health Sciences
                Pathology and Laboratory Medicine
                Pathogens
                Microbial Pathogens
                Fungal Pathogens
                Cryptococcus Gattii
                Biology and Life Sciences
                Mycology
                Fungal Pathogens
                Cryptococcus Gattii
                Biology and Life Sciences
                Organisms
                Eukaryota
                Fungi
                Cryptococcus
                Cryptococcus Neoformans
                Biology and Life Sciences
                Microbiology
                Medical Microbiology
                Microbial Pathogens
                Fungal Pathogens
                Cryptococcus Neoformans
                Medicine and Health Sciences
                Pathology and Laboratory Medicine
                Pathogens
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                Cryptococcus Neoformans
                Biology and Life Sciences
                Mycology
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                Cryptococcus Neoformans
                Biology and Life Sciences
                Organisms
                Eukaryota
                Fungi
                Yeast
                Candida
                Candida Albicans
                Biology and Life Sciences
                Microbiology
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                Medicine and Health Sciences
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                Pathogens
                Microbial Pathogens
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                Candida Albicans
                Biology and Life Sciences
                Mycology
                Fungal Pathogens
                Candida Albicans
                Research and Analysis Methods
                Animal Studies
                Experimental Organism Systems
                Yeast and Fungal Models
                Candida Albicans
                Biology and Life Sciences
                Organisms
                Eukaryota
                Fungi
                Cryptococcus
                Biology and Life Sciences
                Cell Biology
                Cellular Structures and Organelles
                Cell Walls
                Research and Analysis Methods
                Spectrum Analysis Techniques
                Spectrophotometry
                Cytophotometry
                Flow Cytometry
                Biology and Life Sciences
                Physiology
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                Cytokines
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                Developmental Biology
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