Figure - available via license: Creative Commons Attribution 4.0 International
Content may be subject to copyright.
HFF cells labelled with 13C615N2-lysine (dark gray) were submitted to either infection by A. algerae or hypoxic stress. For each time point of both kinetics the labeled cells were combined at 1∶1 ratio with unlabeled HFF cells (light gray) and proteins were extracted. For each sample a three biological replicate was made. The proteins samples were resolved on SDS-PAGE and total proteins lanes were cut in 3 regular pieces (A, B, C) and processed for in-gel digestion with endoproteinase LysC. LC-MS/MS analysis was then performed and mass spectra were analyzed with the MaxQuant software to achieve the relative protein quantification.
Source publication
Intracellular pathogens including bacteria, viruses and protozoa hijack host cell functions to access nutrients and to bypass cellular defenses and immune responses. These strategies have been acquired through selective pressure and allowed pathogens to reach an appropriate cellular niche for their survival and growth. To get new insights on how pa...
Similar publications
Eukaryotic translation initiation factor 3H subunit (EIF3H) is a member of the EIF3 family and exhibits a central role in translation initiation in higher eukaryotes. Although EIF3H expression is upregulated in numerous tumour types, its potential role in human osteosarcoma (OS) has not yet been investigated. In the present study, it was demonstrat...
Invasion and multiplication of the facultative, cytosolic, enteropathogen Shigella flexneri within the colonic epithelial lining leads to an acute inflammatory response, fever and diarrhea. During the inflammatory process, infected cells are subjected to numerous stresses including heat, oxidative stress and genotoxic stress. The evolutionarily con...
Barley Yellow Dwarf Virus (BYDV) is a positive strand RNA virus that lacks the canonical 5' 7-methylguanosine cap and a 3' poly-A tail. Instead, BYDV utilizes a cruciform cap independent translation element (CITE) in its 3'UTR RNA (BYDV-like CITE or BTE) that binds eukaryotic translation initiation factor (eIF) 4F and recruits 40S ribosomal subunit...
Tumor-associated autoantibodies are promising diagnostic biomarkers for early detection of tumors. We have screened a novel tumor-associated autoantibody in hepatocellular carcinoma (HCC) model mice. Its target antigen was identified as eukaryotic translation initiation factor 3 subunit A (EIF3A) by proteomic analysis, and the elevated expression o...
Translation initiation of most mammalian mRNAs is mediated by a 5' cap structure that binds eukaryotic initiation factor 4E (eIF4E). However, inactivation of eIF4E does not impair translation of many capped mRNAs, suggesting an unknown alternate mechanism may exist for cap-dependent but eIF4E-independent translation. We show that DAP5, an eIF4GI ho...
Citations
... The humoral immune system consists of antimicrobial peptide (AMP) production, primarily protecting against bacterial pathogens. For intracellular parasites, insects can clear invading parasites by eliciting oxidative stress [15][16][17] . The intestinal epithelial and macrophage cells produce reactive oxygen species (ROS) 18 , including superoxide anion (O 2 -), hydrogen peroxide (H 2 O 2 ), and hydroxyl radical (HO•). ...
Nosema ceranae is an intracellular parasite invading the midgut of honeybees, which causes serious nosemosis implicated in honeybee colony losses worldwide. The core gut microbiota is involved in protecting against parasitism, and the genetically engineering of the native gut symbionts provides a novel and efficient way to fight pathogens. Here, using laboratory-generated bees mono-associated with gut members, we find that Snodgrassella alvi inhibit microsporidia proliferation, potentially via the stimulation of host oxidant-mediated immune response. Accordingly, N. ceranae employs the thioredoxin and glutathione systems to defend against oxidative stress and maintain a balanced redox equilibrium, which is essential for the infection process. We knock down the gene expression using nanoparticle-mediated RNA interference, which targets the γ-glutamyl-cysteine synthetase and thioredoxin reductase genes of microsporidia. It significantly reduces the spore load, confirming the importance of the antioxidant mechanism for the intracellular invasion of the N. ceranae parasite. Finally, we genetically modify the symbiotic S. alvi to deliver dsRNA corresponding to the genes involved in the redox system of the microsporidia. The engineered S. alvi induces RNA interference and represses parasite gene expression, thereby inhibits the parasitism significantly. Specifically, N. ceranae is most suppressed by the recombinant strain corresponding to the glutathione synthetase or by a mixture of bacteria expressing variable dsRNA. Our findings extend our previous understanding of the protection of gut symbionts against N. ceranae and provide a symbiont-mediated RNAi system for inhibiting microsporidia infection in honeybees.
... ABCD are involved in peroxisomal import of both fatty acids and acylCoAs, whereas PEX19 is a chaperon protein involved in peroxisomal function [38]. The down-regulation of all these genes may contribute to higher cytoplasmic availability of cholesterol and fatty acids, making them available for use by the parasite for its own energy metabolism and membrane development [48][49][50]. In addition, ABC transporters are ATP-dependent and it should be noted that their down-regulation can also reduce ATP expenditure by the cell making more of it available for use by the parasites. ...
... In our study, the observed transcriptional dysregulations also corroborates previous studies on microsporidia infection biology and impact on organisms [9,37,48,49,[75][76][77]. Namely, our data further suggests a role of phosphatidylcholine in microsporidian biology, the downregulation of some immune pathways, and the induction of DNA damage and higher mutation rates in infected cells. ...
Anncaliia algerae belongs to microsporidia, a group of obligate intracellular parasites related to fungi. These parasites are largely spread in water and food-webs and can infect a wide variety of hosts ranging from invertebrates to vertebrates including humans. In humans, microsporidian infections are mainly opportunistic as immunocompetent hosts can clear parasites naturally. Recent studies however have reported persistent microsporidian infections and have highlighted them as a risk factor in colon cancer. This may be a direct result of cell infection or may be an indirect effect of the infectious microenvironment and the host's response. In both cases, this raises the question of the effects of microsporidian infection at the host and host-cell levels.
We aimed to address the question of human host intracellular response to microsporidian infection through a transcriptomic kinetic study of human foreskin fibroblasts (HFF) infected with A.algerae, a human infecting microsporidia with an exceptionally wide host range. We focused solely on host response studying both coding and small non-coding miRNA expression.
Our study revealed a generalized down-regulation of cell miRNAs throughout infection with up to 547 different miRNAs downregulated at some timepoints and also transcriptomic dysregulations that could facilitate parasite development with immune and lipid metabolism genes modulation. We also hypothesize possible small nucleic acid expropriation explaining the miRNA downregulation.
This work contributes to a better understanding of the dialogue that can occur between an intracellular parasite and its host at the cellular level, and can guide future studies on microsporidian infection biology to unravel the mode of action of these minimalist parasites at the tissue or host levels.We have also generated a kinetic and comprehensive transcriptomic data set of an infectious process that can help support comparative studies in the broader field of parasitology. Lastly, these results may warrant for caution regarding microsporidian exposure and persistent infections.
... À ce jour, l'étude simultanée des évènements moléculaires de l'hôte et du champignon n'a pu être réalisée que par l'utilisation d'approches transcriptomiques (Pan et al., 2018a ;Peyraud et al., 2017 ;Puri et al., 2016). Cependant, si ce type d'analyse permet d'apporter de nouveaux éléments sur la compréhension du dialogue moléculaire entre une plante et son pathogène (Gottwald et al., 2012 ;Kugler et al., 2013), il n'offre qu'une information incomplète de l'étendue des régulations cellulaires mises en oeuvre au cours de l'interaction (Chetouhi et al., 2015 ;Panek et al., 2014 ;Quirino et al., 2010). En effet, il est maintenant établi que l'expression des gènes est régulée à différents niveaux et que les changements d'abondances des ARNm ne sont que très faiblement corrélés aux abondances des protéines correspondantes (Maier et al., 2009 ;Nie et al., 2006 ;Ponnala et al., 2014). ...
Les cultures de blé sont régulièrement exposées à une forte pression parasitaire exercée par un cortège complexe d’agents pathogènes capables d’attaquer les différentes parties de la plante. La fusariose de l'épi ou FHB (Fusarium head blight), principalement causée par le champignon ascomycète Fusarium graminearum, représente une préoccupation majeure, car cette maladie affecte non seulement le rendement et la qualité des grains, mais également leur état sanitaire via la production in planta de mycotoxines. À ce jour, les méthodes de lutte reposent sur l’utilisation combinée de pratiques culturales adaptées, de traitements fongicides et de cultivars génétiquement plus résistants. Toutefois, lors d’épisodes climatiques favorables et de fortes pressions parasitaires, ces méthodes s’avèrent incapables de contrôler efficacement le développement d’épidémies dommageables pour les cultures, la raison étant la faible efficacité de la grande majorité des sources de résistance présentes dans les variétés cultivées. Identifier des sources de résistance plus efficaces et surtout plus durables représente aujourd’hui un objectif majeur. Une des alternatives à la recherche de source de résistance classiques repose sur l’identification des facteurs clefs contrôlant la sensibilité de la plante à un agent pathogène, et leur utilisation pour générer des résistances considérées comme potentiellement plus durables. De façon à identifier des gènes de sensibilité du blé au champignon F. graminearum, agent principal de la fusariose de l’épi chez le blé tendre, nous avons mis en place, une stratégie exploratoire à large échelle tenant compte de la diversité, de la dynamique et des spécificités des évènements moléculaires protéiques imputables aux deux protagonistes. Cette stratégie nous a donné accès pour la première fois aux ajustements simultanés des protéomes végétal et fongique au cours des étapes précoces de l’interaction, et plus spécifiquement lors de la transition 48 – 72 hpi. Ces résultats suggèrent que la stratégie fongique s’ajuste de manière dynamique au contexte du développement du grain. De plus, la surreprésentation de motifs d’adressage chloroplastique retrouvée au niveau des séquences protéiques des effecteurs fongiques, associée aux variations d’abondances des protéines du chloroplaste, suggèrent que cet organite pourrait être une des cibles principales des effecteurs fongiques et un élément clé du processus de manipulation parasitaire. Enfin, l’analyse fine du dialogue moléculaire contrôlant le développement de l’interaction entre différents cultivars de blé avec différentes souches fongiques a permis de mettre en évidence l’existence d’un core dual-protéome dont les seules variations quantitatives de ces composantes permettent d’expliquer les différents niveaux de sensibilité ou d’agressivité observés. Ce corpus de résultats permet de proposer la sensibilité comme étant issue de processus moléculaires communs entre différents fonds génétiques de blé et servira de base pour l’élaboration de nouvelles sources de résistance.
... The innate immune response to this microbial colonisation is increased urothelial cell shedding, which may be promoted by mast cells [41]. The shed cells may be colonised or unaffected, but the proportion of parasitised cells in the face of infection would be expected to increase [42]. This sediment, a mixture of white cells, epithelial cells and debris, is likely to collect under the influence of gravity at the bladder base, forming a sampling target that should not be influenced by dilution effects. ...
Introduction and hypothesisMidstream urine (MSU) is key in assessing lower urinary tract syndrome (LUTS), but contingent on some assumptions. The aim of this study was to compare the occurrence of contamination and the quality of substrates obtained from four different collections: MSU, catheter specimen urine (CSU), a commercial MSU collecting device (Peezy) and a natural void. Contamination was quantified by differential, uroplakin-positive, urothelial cell counts.Methods
This was a single blind, crossover study conducted in two phases. First, we compared the MSU with CSU using urine culture, pyuria counts and differential counting of epithelial cells after immunofluorescence staining for uroplakin III (UP3). Second, we compared the three non-invasive (MSU, Peezy MSU™, natural void) methods using UP3 antibody staining only.ResultsThe natural void was best at collecting bladder urinary sediment, with the majority of epithelial cells present derived from the urinary tract. CSU sampling missed much of the urinary sediment and showed sparse culture results. Finally, the MSU collection methods did not capture much of the bladder sediment.Conclusion
We found little evidence for contamination with the four methods. Natural void was the best method for harvesting shed urothelial cells and white blood cells. It provides a richer sample of the inflammatory exudate, including parasitised urothelial cells and the microbial substrate. However, if the midstream sample is believed to be important, the MSU collection device is advantageous.
... This phenomenon may be due to the fact that N. ceranae is highly dependent on host nutritional status for its development, suggesting that the microparasite uses amino acids from its hosts. [61,65]. In the future, controlled laboratory experiments should be conducted to search for a possible mechanism that could be associated with the effects of ABA over Nosema development in honey bees (e.g., bioassays that inoculate bees individually with standardized concentrations). ...
In temperate climates, beekeeping operations suffer colony losses and colony depopulation of Apis mellifera during overwintering, which are associated with biotic and abiotic stressors that impact bees' health. In this work, we evaluate the impacts of abscisic acid (ABA) dietary supplementation on honey bee colonies kept in Langstroth hives. The effects of ABA were evaluated in combination with two different beekeeping nutritional strategies to confront overwintering: "honey management" and "syrup management". Specifically, we evaluated strength parameters of honey bee colonies (adult bee and brood population) and the population dynamics of Nosema (prevalence and intensity) associated with both nutritional systems and ABA supplementation during the whole study (late autumn-winter-early spring). The entire experiment was designed and performed with a local group of beekeepers, "Azahares del sudeste", who showed interest in answering problems associated with the management of honey bee colonies during the winter. The results indicated that the ABA supplementation had positive effects on the population dynamics of the A. mellifera colonies during overwintering and on the nosemosis at colony level (prevalence) in both nutritional strategies evaluated.
... When the superficial bladder cells have been invaded, the uropathogens rapidly begin to replicate inside the bladder cells, forming intracellular communities, also known as intracellular colonisation. Intracellular pathogens such as Escherichia coli hijack bladder cells that line the urothelium, allowing pathogens to reach an appropriate intracellular position for their survival and replication (Panek et al, 2014). ...
... Accordingly, they might contribute to the adaptation to different hosts (Dean 2005;Amyotte et al. 2012;Raffaele and Kamoun 2012). In the case of the microsporidium Anncaliia algerae, these elements might be speculated to help eluding the host immune system by acting as a lure (Panek et al. 2014). ...
Metchnikovellids are highly specialized hyperparasites, which infect and reproduce inside gregarines (Apicomplexa) inhabiting marine invertebrates. Their phylogenetic affiliation was under constant discussion until recently, when analysis of the first near-complete metchnikovellid genome, that of Amphiamblys sp., placed it in a basal position with respect to most Microsporidia. Microsporidia are a highly diversified lineage of extremely reduced parasites related to Rozellida (Rozellosporidia = Rozellomycota = Cryptomycota) within the Holomycota clade of Opisthokonta. By sequencing DNA from a single-isolated infected gregarine cell we obtained an almost complete genome of a second metchnikovellid species, and the first one of a taxonomically described and well-documented species, Metchnikovella incurvata. Our phylogenomic analyses show that, despite being considerably divergent from each other, M. incurvata forms a monophyletic group with Amphiamplys sp., and confirm that metchnikovellids are one of the deep branches of Microsporidia. Comparative genomic analysis demonstrates that, like most Microsporidia, metchnikovellids lack mitochondrial genes involved in energy transduction and are thus incapable of synthesizing their own ATP via mitochondrial oxidative phosphorylation. They also lack the horizontally acquired ATP transporters widespread in most Microsporidia. We hypothesize that a family of mitochondrial carrier proteins evolved to transport ATP from the host into the metchnikovellid cell. We observe the progressive reduction of genes involved in DNA repair pathways along the evolutionary path of Microsporidia, which might explain, at least partly, the extremely high evolutionary rate of the most derived species. Our data also suggest that genome reduction and acquisition of novel genes co-occurred during the adaptation of Microsporidia to their hosts.
... are highly dependent on host nutritional status for their own development, e.g. host amino acids 41,42 , Adenosine triphosphate (ATP) 43,44 or other core nutrients 45 . Therefore, it appears obvious that any individual host with a pollen-rich diet becomes ideal for Nosema spp. ...
Multiple infections are common in honey bees, Apis mellifera, but the possible role of nutrition in this regard is poorly understood. Microsporidian infections, which are promoted by protein-fed, can negatively correlate with virus infections, but the role of protein nutrition for the microsporidian-virus interface is unknown. Here, we challenged naturally deformed wing virus - B (DWV-B) infected adult honey bee workers fed with or without pollen (= protein) in hoarding cages, with the microsporidian Nosema ceranae. Bee mortality was recorded for 14 days and N. ceranae spore loads and DWV-B titers were quantified. Amongst the groups inoculated with N. ceranae, more spores were counted in protein-fed bees. However, N. ceranae infected bees without protein-diet had reduced longevity compared to all other groups. N. ceranae infection had no effect on protein-fed bee's longevity, whereas bees supplied only with sugar-water showed reduced survival. Our data also support that protein-feeding can have a significant negative impact on virus infections in insects. The negative correlation between N. ceranae spore loads and DWV-B titers was stronger expressed in protein-fed hosts. Proteins not only enhance survival of infected hosts, but also significantly shape the microsporidian-virus interface, probably due to increased spore production and enhanced host immunity.
... The RNA-binding ability of IFIT2 protein is required for antiviral activity and to promote apoptosis (Reich, 2013). Further, IFIT2 is also known to modulate the microtubule network implicated in cell reorganization during Anncaliia algerae infection as a host response to clear infection (Panek et al., 2014). The cytokines CCL5 and IL4 are part of the cascade of events leading to efficient parasite control in Leishmania major (Santiago et al., 2004) and Listeria (Kaufmann, Emoto, Szalay, Barsig, & Flesch, 1997) infections, respectively. ...
SUMOylation is a reversible post translational modification of proteins that regulates protein stabilization, nucleocytoplasmic transport and protein-protein interactions. Several viruses and bacteria modulate host SUMOylation machinery for efficient infection. Plasmodium sporozoites are infective forms of malaria parasite that infect hepatocytes and transforms into exoerythrocytic forms (EEFs). Here we show that, during EEF development, the distribution of SUMOylated proteins in host cell nuclei was significantly reduced and expression of the SUMOylation enzymes was downregulated. Plasmodium EEFs destabilized the host cytoplasmic protein SMAD4 by inhibiting its SUMOylation. SUMO1 overexpression was detrimental to EEF growth and insufficiency of the only conjugating enzyme Ubc9/E2 promoted EEF growth. The expression of genes involved in suppression of host cell defense pathways during infection was reversed during SUMO1 overexpression, as revealed by transcriptomic analysis. The inhibition of host cell SUMOylation was also observed in Toxoplasma. We provide a hitherto unknown mechanism of regulating host gene expression through altering host SUMOylation by Apicomplexan parasites.
... This process is illustrated in image 1. 5 Image 1: Urinary tract infection of the bladder (Health Navigator, 2016) When the superficial cells have been invaded, the uropathogens rapidly replicate and form intracellular communities, also known as intracellular colonisation. Intracellular pathogens such as bacteria hijack cells which allow pathogens to reach an appropriate cellular niche for their survival and replication (Panek et al., 2014). Image 2 illustrates this process observed in a murine model which closely resembles the human bladder. ...
A urinary tract infection (UTI) is one of the leading reasons for treatment in primary healthcare. It is estimated that 50% of the female population in the UK will have least one occurrence of the infection in their lifetime. It is a debilitating condition and causes a variety of lower urinary tract symptoms (LUTS). The recommended practice for detecting a UTI is by analysing a urine specimen and culturing the sample for bacterial growth and antibiotic sensitivities. There are two main specimen collection methods: the midstream urine (MSU) and the catheter specimen of urine (CSU). The CSU is recognised as the gold standard, but requires an invasive procedure. The MSU which is the non-invasive clinical standard is regarded as insufficient because the method is frequently reported as contaminated with skin and vaginal flora, but the definitions of contamination in the literature varies. Drawing on the published body of knowledge, this study aimed to investigate and determine what constitutes contamination using microbiological culturing and the uroplakin-3 cell staining technique that detects the presence of cells that originate from the bladder.
A two phase, single blind, cross over design study was conducted, comparing four different urine specimen collection methods. Experiment one tested the hypothesis that a MSU has equal merits to a CSU when capturing urothelial cells that are indicative of a UTI. A total of 60 patients and 30 controls were recruited into the study. The MSU specimens were compared with the CSU specimens to determine urinary cell origin using uroplakin-3 staining. The findings proved that the cells found in the MSU were not contaminants as commonly assumed, but were inflammatory markers of infection invading the lower urinary tract. Experiment two tested the hypothesis that if a MSU has equal merits to a CSU, then a directly voided urine specimen (natural urination) will be the optimal method when capturing the majority of urothelial cells that have been exfoliated from the bladder. A total of 31 patients were recruited and the MSU specimens were compared with the directly voided urine to determine the proportion of cells that originate from the bladder. The findings demonstrated that the directly voided urine was the optimal method and had the ability to capture predominant urothelial cells.
A qualitative study of patient views and experiences of urine specimen collection was conducted. Thirty patients were interviewed and the data were analysed for recurrent themes. The study had shown an ideal urine specimen is that which is sensitive to the underlying pathology of a UTI. It is also a urine specimen that is easy to collect. The direct void is the recommended method of choice but should be accompanied with microscopy. Uroplakin staining should be initiated to further detect the positive presence of uroepithelial cells when distinguishing the difference between urinary contamination.
