scholarly journals Proteinous Components of Neutrophil Extracellular Traps Are Arrested by the Cell Wall Proteins of Candida albicans during Fungal Infection, and Can Be Used in the Host Invasion

Cells ◽  
2021 ◽  
Vol 10 (10) ◽  
pp. 2736
Author(s):  
Justyna Karkowska-Kuleta ◽  
Magdalena Smolarz ◽  
Karolina Seweryn-Ozog ◽  
Dorota Satala ◽  
Marcin Zawrotniak ◽  
...  
Keyword(s):  
Candida Albicans ◽  
Fungal Pathogen ◽  
Defense Mechanisms ◽  
Triosephosphate Isomerase ◽  
Neutrophil Extracellular Traps ◽  
Phosphoglycerate Mutase ◽  
Human Immune System ◽  
Extracellular Traps ◽  
Human Proteins ◽  
Opportunistic Fungal Pathogen

One of defense mechanisms of the human immune system to counteract infection by the opportunistic fungal pathogen Candida albicans is the recruitment of neutrophils to the site of invasion, and the subsequent production of neutrophil extracellular traps (NETs) that efficiently capture and kill the invader cells. In the current study, we demonstrate that within these structures composed of chromatin and proteins, the latter play a pivotal role in the entrapment of the fungal pathogen. The proteinous components of NETs, such as the granular enzymes elastase, myeloperoxidase and lactotransferrin, as well as histones and cathelicidin-derived peptide LL-37, are involved in contact with the surface of C. albicans cells. The fungal partners in these interactions are a typical adhesin of the agglutinin-like sequence protein family Als3, and several atypical surface-exposed proteins of cytoplasmic origin, including enolase, triosephosphate isomerase and phosphoglycerate mutase. Importantly, the adhesion of both the elastase itself and the mixture of proteins originating from NETs on the C. albicans cell surface considerably increased the pathogen potency of human epithelial cell destruction compared with fungal cells without human proteins attached. Such an implementation of adsorbed NET-derived proteins by invading C. albicans cells might alter the effectiveness of the fungal pathogen entrapment and affect the further host colonization.

Host defense mechanisms induce genome instability leading to rapid evolution in an opportunistic fungal pathogen

2021 ◽  
Author(s):  
Amanda Smith ◽  
Levi Morran ◽  
Meleah A. Hickman
Keyword(s):  
Fungal Pathogen ◽  
Defense Mechanisms ◽  
Large Scale ◽  
Genome Instability ◽  
Rapid Evolution ◽  
Genomic Changes ◽  
Opportunistic Fungal Pathogen ◽  
Immunocompromised Hosts ◽  
Stressful Environments

The ability to generate genetic variation facilitates rapid adaptation in stressful environments. The opportunistic fungal pathogen Candida albicans frequently undergoes large-scale genomic changes, including aneuploidy and loss-of heterozygosity (LOH), following exposure to host environments. However, the specific host factors inducing C. albicans genome instability remain largely unknown. Here, we leveraged the genetic tractability of nematode hosts to investigate whether innate immune components, including antimicrobial peptides (AMPs) and reactive oxygen species (ROS), induced host-associated C. albicans genome instability. C. albicans associated with immunocompetent hosts carried multiple large-scale genomic changes including LOH, whole chromosome, and segmental aneuploidies. In contrast, C. albicans associated with immunocompromised hosts deficient in AMPs or ROS production had reduced LOH frequencies and fewer, if any, additional genomic changes. To evaluate if extensive host-induced genomic changes had long-term consequences for C. albicans adaptation, we experimentally evolved C. albicans in either immunocompetent or immunocompromised hosts and selected for increased virulence. C. albicans evolved in immunocompetent hosts rapidly increased virulence, but not in immunocompromised hosts. Taken together, this work suggests that host-produced ROS and AMPs induces genotypic plasticity in C. albicans which facilitates rapid evolution.

scholarly journals Inactivation of α1-proteinase inhibitor by Candida albicans aspartic proteases favors the epithelial and endothelial cell colonization in the presence of neutrophil extracellular traps.

2015 ◽  
Vol 63 (1) ◽  
Author(s):  
Mariusz Gogol ◽  
Dominika Ostrowska ◽  
Kinga Klaga ◽  
Oliwia Bochenska ◽  
Natalia Wolak ◽  
...  
Keyword(s):  
Candida Albicans ◽  
Proteinase Inhibitor ◽  
Proteolytic Enzymes ◽  
Fungal Infections ◽  
Critical Role ◽  
Neutrophil Extracellular Traps ◽  
Host Cells ◽  
Inhibition Mechanism ◽  
Aspartic Proteases ◽  
Extracellular Traps

Candida albicans, a causative agent of opportunistic fungal infections in immunocompromised patients, uses ten secreted aspartic proteases (SAPs) to deregulate the homeostasis of the host organism on many levels. One of these deregulation mechanisms involves a SAP-dependent disturbance of the control over proteolytic enzymes of the host by a system of dedicated proteinase inhibitors, with one important example being the neutrophil elastase and alpha1-proteinase inhibitor (A1PI). In this study, we found that soluble SAPs 1-4 and the cell membrane-anchored SAP9 efficiently cleaved A1PI, with the major cleavage points located at the C-terminal part of A1PI in a close vicinity to the reactive-site loop that plays a critical role in the inhibition mechanism. Elastase is released by neutrophils to the environment during fungal infection through two major processes, a degranulation or formation of neutrophil extracellular traps (NET). Both, free and NET-embedded elastase forms, were found to be controlled by A1PI. A local acidosis, resulting from the neutrophil activity at the infection sites, favors A1PI degradation by SAPs. The deregulation of NET-connected elastase affected a NET-dependent damage of epithelial and endothelial cells, resulting in the increased susceptibility of these host cells to candidal colonization. Moreover, the SAP-catalyzed cleavage of A1PI was found to decrease its binding affinity to a proinflammatory cytokine, interleukin-8. The findings presented here suggest a novel strategy used by C. albicans for the colonization of host tissues and overcoming the host defense.

scholarly journals High-Throughput Nano-Biofilm Microarray for Antifungal Drug Discovery

mBio ◽  
2013 ◽  
Vol 4 (4) ◽  
Author(s):  
Anand Srinivasan ◽  
Kai P. Leung ◽  
Jose L. Lopez-Ribot ◽  
Anand K. Ramasubramanian
Keyword(s):  
Cell Culture ◽  
Candida Albicans ◽  
Drug Discovery ◽  
High Throughput ◽  
Fungal Pathogen ◽  
Microarray Platform ◽  
Microbial Cell ◽  
Content Type ◽  
Petri Dishes ◽  
Opportunistic Fungal Pathogen

ABSTRACT Micro- and nanoscale technologies have radically transformed biological research from genomics to tissue engineering, with the relative exception of microbial cell culture, which is still largely performed in microtiter plates and petri dishes. Here, we present nanoscale culture of the opportunistic fungal pathogen Candida albicans on a microarray platform. The microarray consists of 1,200 individual cultures of 30 nl of C. albicans biofilms (“nano-biofilms”) encapsulated in an inert alginate matrix. We demonstrate that these nano-biofilms are similar to conventional macroscopic biofilms in their morphological, architectural, growth, and phenotypic characteristics. We also demonstrate that the nano-biofilm microarray is a robust and efficient tool for accelerating the drug discovery process: (i) combinatorial screening against a collection of 28 antifungal compounds in the presence of immunosuppressant FK506 (tacrolimus) identified six drugs that showed synergistic antifungal activity, and (ii) screening against the NCI challenge set small-molecule library identified three heretofore-unknown hits. This cell-based microarray platform allows for miniaturization of microbial cell culture and is fully compatible with other high-throughput screening technologies. IMPORTANCE Microorganisms are typically still grown in petri dishes, test tubes, and Erlenmeyer flasks in spite of the latest advances in miniaturization that have benefitted other allied research fields, including genomics and proteomics. Culturing microorganisms in small scale can be particularly valuable in cutting down time, cost, and reagent usage. This paper describes the development, characterization, and application of nanoscale culture of an opportunistic fungal pathogen, Candida albicans. Despite a more than 2,000-fold reduction in volume, the growth characteristics and drug response profiles obtained from the nanoscale cultures were comparable to the industry standards. The platform also enabled rapid identification of new drug candidates that were effective against C. albicans biofilms, which are a major cause of mortality in hospital-acquired infections.

The hematoregulatory peptide, SK&F 107647, in combination with antifungal therapy in murine Candida albicans infections

1995 ◽  
Vol 73 (S1) ◽  
pp. 1199-1205 ◽  
Author(s):  
Peter L. DeMarsh ◽  
Scott K. Sucoloski ◽  
Carrie L. Frey ◽  
Louis M. Pelus ◽  
Pradip K. Bhatnagar ◽  
...  
Keyword(s):  
Candida Albicans ◽  
Amphotericin B ◽  
Antifungal Therapy ◽  
Fungal Pathogen ◽  
Prophylactic Treatment ◽  
Lethal Dose ◽  
Survival Rates ◽  
Low Molecular Weight ◽  
Opportunistic Fungal Pathogen ◽  
Immunosuppressed Mice

SK&F 107647, a novel synthetic low molecular weight peptide, has been shown to be a potent hematoregulatory agent. We have previously demonstrated that SK&F 107647 administration can prolong survival in both immunosuppressed and normal mice challenged with the opportunistic fungal pathogen Candida albicans. Additionally, we have determined the effect of prophylactic SK&F 107647 treatment combined with conventional antifungal therapy on the survival of mice challenged with a lethal dose of C. albicans. Prophylactic treatment with SK&F 107647 or therapeutic treatment with the antifungals fluconazole or amphotericin B significantly increased the survival rates of immunosuppressed mice challenged with a lethal dose of C. albicans. However, the combination of SK&F 107647 treatment followed by antifungal therapy resulted in statistically significant increases in survival over that observed with either therapy alone. These results indicated that the hematoregulatory factor(s) elicited by SK&F 107647 enhance the survival of mice treated with conventional therapies in a model of experimental systemic candidiasis. Key words: SK&F 107647, Candida albicans, hematoregulatory, fluconazole, amphotericin B.

scholarly journals Rac1 Dynamics in the Human Opportunistic Fungal Pathogen Candida albicans

PLoS ONE ◽  
2010 ◽  
Vol 5 (10) ◽  
pp. e15400 ◽  
Author(s):  
Romain Vauchelles ◽  
Danièle Stalder ◽  
Thomas Botton ◽  
Robert A. Arkowitz ◽  
Martine Bassilana
Keyword(s):  
Candida Albicans ◽  
Fungal Pathogen ◽  
Opportunistic Fungal Pathogen

scholarly journals Ascorbic Acid Inhibition of Candida albicans Hsp90-Mediated Morphogenesis Occurs via the Transcriptional Regulator Upc2

2014 ◽  
Vol 13 (10) ◽  
pp. 1278-1289 ◽  
Author(s):  
Frédérique Van Hauwenhuyse ◽  
Alessandro Fiori ◽  
Patrick Van Dijck
Keyword(s):  
Ascorbic Acid ◽  
Candida Albicans ◽  
Fungal Pathogen ◽  
Transcriptional Regulator ◽  
Hsp90 Inhibitor ◽  
Ergosterol Biosynthesis ◽  
Content Type ◽  
Temperature Changes ◽  
Opportunistic Fungal Pathogen ◽  
Hsp90 Function

ABSTRACTMorphogenetic transitions of the opportunistic fungal pathogenCandida albicansare influenced by temperature changes, with induction of filamentation upon a shift from 30 to 37°C. Hsp90 was identified as a major repressor of an elongated cell morphology at low temperatures, as treatment with specific inhibitors of Hsp90 results in elongated growth forms at 30°C. Elongated growth resulting from a compromised Hsp90 is considered neither hyphal nor pseudohyphal growth. It has been reported that ascorbic acid (vitamin C) interferes with the yeast-to-hypha transition inC. albicans. In the present study, we show that ascorbic acid also antagonizes the morphogenetic change caused by hampered Hsp90 function. Further analysis revealed that Upc2, a transcriptional regulator of genes involved in ergosterol biosynthesis, and Erg11, the target of azole antifungals, whose expression is in turn regulated by Upc2, are required for this antagonism. Ergosterol levels correlate with elongated growth and are reduced in cells treated with the Hsp90 inhibitor geldanamycin (GdA) and restored by cotreatment with ascorbic acid. In addition, we show that Upc2 appears to be required for ascorbic acid-mediated inhibition of the antifungal activity of fluconazole. These results identify Upc2 as a major regulator of ascorbic acid-induced effects inC. albicansand suggest an association between ergosterol content and elongated growth upon Hsp90 compromise.

scholarly journals Extracellular Nucleic Acids Present in the Candida albicans Biofilm Trigger the Release of Neutrophil Extracellular Traps

2021 ◽  
Vol 11 ◽  
Author(s):  
Magdalena Smolarz ◽  
Marcin Zawrotniak ◽  
Dorota Satala ◽  
Maria Rapala-Kozik
Keyword(s):  
Candida Albicans ◽  
Nucleic Acids ◽  
Fungal Infection ◽  
Fungal Infections ◽  
Neutrophil Extracellular Traps ◽  
Human Neutrophils ◽  
High Concentration ◽  
First Line ◽  
Extracellular Traps ◽  
Biofilm Structure

Neutrophils, the first line of the host’s defense, use a variety of antimicrobial mechanisms to fight invading pathogens. One of the most crucial is the production of neutrophil extracellular traps (NETs) in the process called NETosis. The unique structure of NETs effectively inhibits the spread of pathogens and ensures their exposure to a high concentration of NET-embedded antimicrobial compounds. NETosis strategy is often used by the host to defend against fungal infection caused by Candida albicans. In immunocompromised patients, this microorganism is responsible for developing systemic fungal infections (candidiasis). This is correlated with the use of a vast array of virulence factors, leading to the acquisition of specific resistance to host defense factors and available drug therapies. One of the most important features favoring the development of drug resistance is a C. albicans ability to form biofilms that protect fungal cells mainly through the production of an extracellular matrix (ECM). Among the main ECM-building macromolecules extracellular nucleic acids have been identified and their role is probably associated with the stbilization of the biofilm structure. The complex interactions of immune cells with the thick ECM layer, comprising the first line of contact between these cells and the biofilm structure, are still poorly understood. Therefore, the current studies aimed to assess the release of extracellular nucleic acids by C. albicans strains at different stages of biofilm formation, and to determine the role of these molecules in triggering the NETosis. We showed for the first time that fungal nucleic acids, purified directly from mature C. albicans biofilm structure or obtained from the whole fungal cells, have the potential to induce NET release in vitro. In this study, we considered the involvement of TLR8 and TLR9 in NETosis activation. We showed that DNA and RNA molecules initiated the production of reactive oxygen species (ROS) by activation of the NADPH oxidase complex, essential for ROS-dependent NETosis. Furthermore, analysis of the cell migration showed that the nucleic acids located in the extracellular space surrounding the biofilm may be also effective chemotactic factors, driving the dynamic migration of human neutrophils to the site of ongoing fungal infection.

scholarly journals Plasma Membrane Potential of Candida albicans Measured by Di-4-ANEPPS Fluorescence Depends on Growth Phase and Regulatory Factors

2019 ◽  
Vol 7 (4) ◽  
pp. 110 ◽  
Author(s):  
Suchodolski ◽  
Krasowska
Keyword(s):  
Candida Albicans ◽  
Plasma Membrane ◽  
Membrane Potential ◽  
Cell Membrane ◽  
Growth Phase ◽  
Fungal Pathogen ◽  
Physiological State ◽  
External Factors ◽  
Good Tool ◽  
Opportunistic Fungal Pathogen

The potential of the plasma membrane (Δѱ) regulates the electrochemical potential between the outer and inner sides of cell membranes. The opportunistic fungal pathogen, Candida albicans, regulates the membrane potential in response to environmental conditions, as well as the physiological state of the cell. Here we demonstrate a new method for detection of cell membrane depolarization/permeabilization in C. albicans using the potentiometric zwitterionic dye di-4-ANEPPS. Di-4-ANEPPS measures the changes in the cell Δѱ depending on the phases of growth and external factors regulating Δѱ, such as potassium or calcium chlorides, amiodarone or DM-11 (inhibitor of H+-ATPase). We also demonstrated that di-4-ANEPPS is a good tool for fast measurement of the influence of amphipathic compounds on Δѱ.

scholarly journals Drosophila melanogaster Thor and Response to Candida albicans Infection

2007 ◽  
Vol 6 (4) ◽  
pp. 658-663 ◽  
Author(s):  
A. Levitin ◽  
A. Marcil ◽  
G. Tettweiler ◽  
M. J. Laforest ◽  
U. Oberholzer ◽  
...  
Keyword(s):  
Innate Immunity ◽  
Drosophila Melanogaster ◽  
Candida Albicans ◽  
Host Defense ◽  
Translational Control ◽  
Fungal Pathogen ◽  
Transcriptional Profiling ◽  
S2 Cells ◽  
Candida Albicans Infection ◽  
Opportunistic Fungal Pathogen

ABSTRACT We used Drosophila melanogaster macrophage-like Schneider 2 (S2) cells as a model to study cell-mediated innate immunity against infection by the opportunistic fungal pathogen Candida albicans. Transcriptional profiling of S2 cells coincubated with C. albicans cells revealed up-regulation of several genes. One of the most highly up-regulated genes during this interaction is the D. melanogaster translational regulator 4E-BP encoded by the Thor gene. Analysis of Drosophila 4E-BP null mutant survival upon infection with C. albicans showed that 4E-BP plays an important role in host defense, suggesting a role for translational control in the D. melanogaster response to C. albicans infection.

Sign in / Sign up

Export Citation Format

Share Document