Autophagy is a highly conserved catabolic pathway that is vital for cells; however, the effects of autophagy on the biofilm formation and antifungal resistance of Candida albicans are still unknown. ...In this study, the potential molecular mechanisms of autophagy in biofilm formation and antifungal resistance were investigated. It was found that 3536 genes were differentially expressed between biofilm and planktonic C. albicans. ATG gene expression and autophagy activity were higher in biofilm than in planktonic C. albicans. Autophagic activities were higher in matured biofilms than that in pre-matured biofilms. Autophagy was involved in C. albicans biofilm formation and its activity increased during biofilm maturation. Further, ALP activity, AO staining cells, and autophagosomes inside cells were obviously reduced in biofilms of atg13Δ/Δ and atg27Δ/Δ strains; moreover, biofilm formation and antifungal resistance were also significantly decreased. Lastly, autophagy regulates biofilm formation and drug resistance of C. albicans and could be served as a new molecular target to the C. albicans biofilm infections.
Glucocorticoid‐induced leucine zipper (GILZ) exerts anti‐inflammatory effects on the immune cells. However, less is known about GILZ function in neutrophils. We aimed to define the specific role of ...GILZ in basal neutrophil activity during an inflammatory response. GILZ knockdown resulted in a persistent activation state of neutrophils, as evidenced by increased phagocytosis, killing activity, and oxidative burst in GILZ‐knockout (KO) neutrophils. This enhanced response caused severe disease in a dinitrobenzene sulfonic acid (DNBS)‐induced colitis model, where GILZ‐KO mice had prominent granulocytic infiltrate and excessive inflammatory state. We used a Candida albicans intraperitoneal infection model to unravel the intracellular pathways affected by GILZ expression in activated neutrophils. GILZ‐KO neutrophils had stronger ability to clear the infectious agent than the wild‐type (WT) neutrophils, and there was more activation of the NOX2 (NADPH oxidase 2) and p47phox proteins, which are directly involved in oxidative burst. Similarly, the MAPK pathway components, that is, ERK and p38, which are involved in the oxidative burst pathway, were highly phosphorylated in GILZ‐KO neutrophils. Evaluation of GILZ expression kinetics during C. albicans infection revealed down‐regulation that correlated inversely with the state of neutrophil activation, which was evaluated as oxidative burst. Overall, our findings define GILZ as a regulator of neutrophil functions, as its expression contributes to limiting neutrophil activation by reducing the activation of the signaling pathways that control the basal neutrophil functions. Controlling GILZ expression could help regulate a continuous inflammatory state that can result in chronic inflammatory and autoimmune diseases.
GILZ expression contributes to the inhibition of neutrophil activation by reducing MAPK pathway protein and NOX2 activity that control basal neutrophil functions.
•The killing process of macrophages to C. albicans could be divided into three stages (early, mid, and late stage).•Autophagy pathway in macrophages does not contribute to anti-Candida at ...mid-stage.•LAP phago-lysosomal fusion obstruction dampens the anti-Candida capability of macrophages at the mid-stage.
In innate immunity, macrophages play critical roles in defending against pathogens via the lysosomal degradation function of autophagy. Two distinct autophagy pathways have been identified in decades: canonical autophagy (referred to as autophagy) and LC3-associated phagocytosis (LAP). Since several conflicting findings about the anti-Candida capability of autophagy (or LAP) have been reported, they serve as the foe or friend for Candida survival is still unclearly. The current study showed that the fungicidal process of THP-1-derived macrophages (THP-1-MФ) against Candida albicans is divided into three stages as follows, the early stage (the first 12 h, increasing in the killing capability), the mid-stage (12–24 h, no change in killing capability), and the late stage (24–48 h, decreasing of the killing capability). Autophagic protein LC3B-II reached the peak in THP-1-MФ after 24 h inoculated either with C.albicans or whole glucan particles (WGP). Thus, both anti-Candida roles of autophagy and the LAP pathway have been detected at the mid-stage. For autophagy, after 24 h inoculation with C.albicans, ULK1 increased, but p-ATG13(s318) decreased obviously in THP-1-MФ, and the killing assay showed that autophagy is unhelpful for Candida killing capability. For the LAP pathway, Rubicon and ROS raised significantly in THP-1-MФ after 24 h inoculated with C.albicans; each inhibition would sharply cut down the LC3B-II accumulation, which indicated that LAP had been induced. However, mCherry-GFP-LC3 fluorescent assay exhibited that LAP phago-lysosomal fusion has been blocked, and Rubicon knockdown facilitated the Candida killing activity. These data indicated that autophagy presented as redundant to Candida defense, and LAP phago-lysosomal fusion obstruction impairs the Candida killing capability of THP-1-MФ at the mid-stage. That may explain the no change in Candida killing capability at the mid-stage.
Candida albicans ( C. albicans ), a microbe commonly isolated from Candida vaginitis patients with vaginal tract infections, transforms from yeast to hyphae and produces many toxins, adhesins, and ...invasins, as well as C. albicans biofilms resistant to antifungal antibiotic treatment. Effective agents against this pathogen are urgently needed. Antimicrobial peptides (AMPs) have been used to cure inflammation and infectious diseases. In this study, we isolated whole housefly larvae insect SVWC peptide 1 (WHIS1), a novel insect single von Willebrand factor C-domain protein (SVWC) peptide from whole housefly larvae. The expression pattern of WHIS1 showed a response to the stimulation of C. albicans . In contrast to other SVWC members, which function as antiviral peptides, interferon (IFN) analogs or pathogen recognition receptors (PRRs), which are the prokaryotically expressed MdWHIS1 protein, inhibit the growth of C. albicans . Eukaryotic heterologous expression of WHIS1 inhibited C. albicans invasion into A549 and HeLa cells. The heterologous expression of WHIS1 clearly inhibited hyphal formation both extracellularly and intracellularly. Furthermore, the mechanism of WHIS1 has demonstrated that it downregulates all key hyphal formation factors ( ALS1, ALS3, ALS5, ECE1, HWP1, HGC1, EFG1 , and ZAP1 ) both extracellularly and intracellularly. These data showed that heterologously expressed WHIS1 inhibits C. albicans invasion into epithelial cells by affecting hyphal formation and adhesion factor-related gene expression. These findings provide new potential drug candidates for treating C. albicans infection.
Candida albicans
is the most common cause of fungal infection. The emergence of drug resistance leads to the need for novel antifungal agents. We aimed to design naphthofuranquinone analogs to treat ...drug-resistant
C. albicans
for topical application on cutaneous candidiasis. The time-killing response, agar diffusion, and live/dead assay of the antifungal activity were estimated against 5-fluorocytosine (5-FC)- or fluconazole-resistant strains. A total of 14 naphthofuranquinones were compared for their antifungal potency. The lead compounds with hydroxyimino (TCH-1140) or
O
-acetyl oxime (TCH-1142) moieties were the most active agents identified, showing a minimum inhibitory concentration (MIC) of 1.5 and 1.2 μM, respectively. Both compounds were superior to 5-FC and fluconazole for killing planktonic fungi. Naphthofuranquinones efficiently diminished the microbes inside and outside the biofilm. TCH-1140 and TCH-1142 were delivered into
C. albicans
-infected keratinocytes to eradicate intracellular fungi. The compounds did not reduce the
C. albicans
burden inside the macrophages, but the naphthofuranquinones promoted the transition of fungi from the virulent hypha form to the yeast form. In the
in vivo
skin mycosis mouse model, topically applied 5-FC and TCH-1140 reduced the
C. albicans
load from 1.5 × 10
6
to 5.4 × 10
5
and 1.4 × 10
5
CFU, respectively. The infected abscess diameter was significantly decreased by TCH-1140 (3-4 mm) as compared to the control (8 mm). The disintegrated skin-barrier function induced by the fungi was recovered to the baseline by the compound. The data support the potential of TCH-1140 as a topical agent for treating drug-resistant
C. albicans
infection without causing skin irritation.
Active tuberculosis patients are at high risk of coinfection with opportunistic fungal pathogen Candida albicans. However, the molecular mechanisms that orchestrate pathogenesis of Mycobacterium ...tuberculosis (Mtb)‐C. albicans coinfection remain elusive. In the current study, we utilize a mouse model to demonstrate that Mtb promotes a macrophage environment that is conducive for C. albicans survival. Mtb‐dependent protein kinase Cζ‐WNT signalling axis induces expression of an E3 ubiquitin ligase, constitutive photomorphogenesis protein 1 (COP1). A secondary infection of C. albicans in such Mtb‐infected macrophages causes COP1 to mediate the proteasomal degradation of interferon regulatory factor 9 (IRF9), a cardinal factor that we identified to arbitrate an inflammatory programmed cell death, pyroptosis. In vivo experiments mimicking a pre‐existing Mtb infection demonstrate that inhibition of pyroptosis in mice results in increased C. albicans burden and aberrant lung tissue architecture, leading to increased host mortality. Together, our study reveals the crucial role of pyroptosis regulation for manifesting a successful C. albicans–Mtb coinfection.
Co‐infection with the opportunistic fungal pathogen Candida albicans commonly occurs in tuberculosis (Mycobacterium tuberculosis, Mtb) patients. In this study, Bhatt et al. established a mouse model to explore the immune and signalling dynamics of co‐infection with Mtb and C. albicans. They show that Candida infection induces pyroptosis in an IRF9‐dependent manner, and this is modulated by pre‐existing Mtb infection, as follows: Mtb‐driven PKCζ‐WNT signalling induces the expression of E3 ubiquitin ligase COP1, which mediates the proteasomal degradation of IRF9. Collectively, their findings indicate that pyroptosis regulation plays a key role in exacerbating the pathogenesis of C. albicans during active tuberculosis.
Ras proteins are highly conserved small GTPases in eukaryotes. GTP-bound Ras binds to effectors to trigger signaling cascades. In order to understand how extensive is the functional homology between ...the highly homologous proteins, S. cerevisiae Ras2 and C. albicans Ras1, we examined whether ScRas2 could functionally complement CaRas1 in activating hyphal morphogenesis as well as GPI anchor biosynthesis. We show that ScRas2 functionally complements CaRas1 in rescuing growth as well as activating hyphal growth, a process that involves plasma membrane localized Ras activating cAMP/PKA signaling via Cyr1. However, ScRas2 is unable to activate the GPI-N-acetylglucosaminyl transferase (GPI-GnT) which catalyzes the first step of GPI biosynthesis. That CaRas1 alone activates GPI-GnT and not ScRas2 suggests that this process is cAMP independent. Interestingly, CaRas1 transcriptionally activates CaGPI2, encoding a GPI-GnT subunit that has been shown to interact with CaRas1 physically. In turn, CaGPI2 downregulates CaGPI19, encoding another GPI-GnT subunit. This has direct consequences for expression of CaERG11, encoding the target of azole antifungals. This effect too is specific to CaRas1 and ScRas2 is unable to replicate it.
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•ScRas2 functionally complements CaRas1 in activating hyphal growth.•But ScRas2 cannot activate the GPI-N-acetylglucosaminyl transferase (GPI-GnT).•That CaRas1, not ScRas2, activates GPI-GnT suggests that this is cAMP independent.•CaRas1 alone activates CaGPI2, a GPI-GnT subunit that it physically interacts with.•CaRas1 affects response to azoles. ScRas2 is unable to replicate this effect.
In recent decades, infections caused by the opportunistic fungus
Candida albicans
have increased, especially in patients with immunodeficiency. In this study, we investigated the mechanism of action ...of sanguinarine (SAN) against
C. albicans
both
in vitro
and
in vivo
. SAN exhibited antifungal activity against
C. albicans
clinical isolates, with MICs in the range of 112.8–150.5 μM. Furthermore, scanning electron and transmission electron microscopy showed that SAN induced morphological changes as well as structure disruption in
C. albicans
cells, including masses of cellular debris, ruptured cell walls, and membrane deformation. Flow cytometry revealed that SAN could lead to cell membrane damage, and ergosterol content analysis indicated that SAN could cause ergosterol content reduction exceeding 90%. Further, we validated the efficacy of SAN against candidiasis caused by
C. albicans
in a murine model, and SAN significantly improved survival and reduced weight loss compared to vehicle. The treatment of 1.5 and 2.5 mg/kg/d SAN obviously reduced the fungal burden in the kidney. In addition, histopathological examination indicated that no fungal cells were observed in lung and kidney tissues after SAN treatment. Hence, this study suggests that SAN is a promising plant-derived compound for the development of an effective anticandidal agent.
Neutrophil extracellular traps (NETs) are a combination of DNA fibers and granular enzymes, such as elastase and myeloperoxidase. In this study, we demonstrate that Candida albicans hyphal (CAH) ...cells and yeast (CAY) cells induce differential amounts, kinetics and mechanisms of NET release. CAH cells induced larger quantities of NET compared to CAY cells and can stimulate rapid NET formation up to 4 h of incubation. CAY cells are, also, able to induce rapid NET formation, but this ability was lost at 4 h. Both reactive oxygen species (ROS) and autophagy are implicated in NET induced by CAH and CAY cells, but with a time-different participation of these two mechanisms. In particular, in the early phase (15 min) CAH cells stimulate NET via autophagy, but not via ROS, while CAY cells induce NET via both autophagy and ROS. At 4 h, only CAH cells stimulate NET formation using autophagy as well as ROS. Finally, we demonstrate that NET release, in response to CAH cells, involves NF-κB activation and is strongly implicated in hyphal destruction.
Vulvovaginal candidiasis (VVC) is a symptomatic inflammation of the vagina mainly caused by
C. albicans
. Other species, such as
C. parapsilosis
,
C. glabrata
,
C. tropicalis
, and
C. krusei
, are ...mainly associated to the recurrent form of the disease (RVVC), although with a lower frequency. In its yeast form,
C. albicans
is tolerated by the vaginal epithelium, but switching to the invasive hyphal form, co-regulated with the expression of genes encoding virulence factors such as secreted aspartyl proteases (Sap) and candidalysin, allows for tissue damage. Vaginal epithelial cells play an important role by impairing
C. albicans
tissue invasion through several mechanisms such as epithelial shedding, secretion of mucin and strong interepithelial cell connections. However, morphotype switching coupled to increasing of the fungal burden can overcome the tolerance threshold and trigger an intense inflammatory response. Pathological inflammation is believed to be facilitated by an altered vaginal microbiome, i.e.,
Lactobacillus
dysbiosis. Notwithstanding the damage caused by the fungus itself, the host response to the fungus plays an important role in the onset of VVC, exacerbating fungal-mediated damage. This response can be triggered by host PRR-fungal PAMP interaction and other more complex mechanisms (i.e., Sap-mediated NLRP3 activation and candidalysin), ultimately leading to strong neutrophil recruitment. However, recruited neutrophils appear to be ineffective at reducing fungal burden and invasion; therefore, they seem to contribute more to the symptoms associated with vaginitis than to protection against the disease. Recently, two aspects of the vulvovaginal environment have been found to associate with VVC and induce neutrophil anergy
in vitro
: perinuclear anti-neutrophil cytoplasmic antibodies (pANCA) and heparan sulfate. Interestingly, CAGTA antibodies have also been found with higher frequency in VVC as compared to asymptomatic colonized women. This review highlights and discusses recent advances on understanding the VVC pathogenesis mechanisms as well as the role of host defenses during the disease.
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