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Nucleic Acids Research Dec 2023Cell autonomous responses to intracellular bacteria largely depend on reorganization of gene expression. To gain isoform-level resolution of these modes of regulation,...
Cell autonomous responses to intracellular bacteria largely depend on reorganization of gene expression. To gain isoform-level resolution of these modes of regulation, we combined long- and short-read transcriptomic analyses of the response of intestinal epithelial cells to infection by the foodborne pathogen Listeria monocytogenes. Among the most striking isoform-based types of regulation, expression of the cellular stress response regulator CIRBP (cold-inducible RNA-binding protein) and of several SRSFs (serine/arginine-rich splicing factors) switched from canonical transcripts to nonsense-mediated decay-sensitive isoforms by inclusion of 'poison exons'. We showed that damage to host cell membranes caused by bacterial pore-forming toxins (listeriolysin O, perfringolysin, streptolysin or aerolysin) led to the dephosphorylation of SRSFs via the inhibition of the kinase activity of CLK1, thereby driving CIRBP alternative splicing. CIRBP isoform usage was found to have consequences on infection, since selective repression of canonical CIRBP reduced intracellular bacterial load while that of the poison exon-containing isoform exacerbated it. Consistently, CIRBP-bound mRNAs were shifted towards stress-relevant transcripts in infected cells, with increased mRNA levels or reduced translation efficiency for some targets. Our results thus generalize the alternative splicing of CIRBP and SRSFs as a common response to biotic or abiotic stresses by extending its relevance to the context of bacterial infection.
Topics: Humans; Alternative Splicing; Listeriosis; Protein Isoforms; RNA-Binding Proteins; Listeria monocytogenes
PubMed: 37941135
DOI: 10.1093/nar/gkad1033 -
Frontiers in Immunology 2024Previous studies have demonstrated an association between lymphatic vessels and diseases caused by bacterial infections. (LM) bacterial infection can affect multiple...
Previous studies have demonstrated an association between lymphatic vessels and diseases caused by bacterial infections. (LM) bacterial infection can affect multiple organs, including the intestine, brain, liver and spleen, which can be fatal. However, the impacts of LM infection on morphological and functional changes of lymphatic vessels remain unexplored. In this study, we found that LM infection not only induces meningeal and mesenteric lymphangiogenesis in mice, but also impairs meningeal lymphatic vessels (MLVs)-mediated macromolecules drainage. Interestingly, we found that the genes associated with lymphatic vessel development and function, such as and , were downregulated, suggesting that LM infection may affect cellular polarization and valve development. On the other hand, photodynamic ablation of MLVs exacerbated inflammation and bacterial load in the brain of mice with LM infection. Overall, our findings indicate that LM infection induces lymphangiogenesis and may affect cell polarization, cavity formation, and valve development during lymphangiogenesis, ultimately impairing MLVs drainage.
Topics: Animals; Mice; Listeria monocytogenes; Listeriosis; Lymphatic Vessels; Lymphangiogenesis; Meninges
PubMed: 38638427
DOI: 10.3389/fimmu.2024.1382971 -
Foods (Basel, Switzerland) Apr 2024is the causative agent of listeriosis, a severe foodborne illness characterized by septicemia, meningitis, encephalitis, abortions, and occasional death in infants and...
is the causative agent of listeriosis, a severe foodborne illness characterized by septicemia, meningitis, encephalitis, abortions, and occasional death in infants and immunocompromised individuals. is composed of four genetic lineages (I, II, III, and IV) and fourteen serotypes. The aim of the current study was to identify proteins that can serve as biomarkers for detection of genetic lineage III strains based on simple antibody-based methods. Liquid chromatography (LC) with electrospray ionization tandem mass spectrometry (ESI MS/MS) followed by bioinformatics and computational analysis were performed on three strains (NRRL B-33007, NRRL B-33014, and NRRL B-33077), which were used as reference strains for lineages I, II, and III, respectively. Results from ESI MS/MS revealed 42 unique proteins present in NRRL B-33077 and absent in NRRL B-33007 and NRRL B-33014 strains. BLAST analysis of the 42 proteins against a broader panel of >80 sequenced strains from lineages I and II revealed four proteins [TM2 domain-containing protein (NRRL B-33077_2770), DUF3916 domain-containing protein (NRRL B-33077_1897), DNA adenine methylase (NRRL B-33077_1926), and protein RhsA (NRRL B-33077_1129)] that have no homology with any sequenced strains in lineages I and II. The four genes that encode these proteins were expressed in strain DE3 and purified. Polyclonal antibodies were prepared against purified recombinant proteins. ELISA using the polyclonal antibodies against 12 lineage I, II, and III isolates indicated that TM2 protein and DNA adenine methylase (Dam) detected all lineage III strains with no reaction to lineage I and II strains. In conclusion, two proteins including TM2 protein and Dam are potentially useful biomarkers for detection and differentiation of lineage III strains in clinical, environmental, and food processing facilities. Furthermore, these results validate the approach of using a combination of proteomics and bioinformatics to identify useful protein biomarkers.
PubMed: 38731673
DOI: 10.3390/foods13091302 -
Frontiers in Microbiology 2023Foodborne illness outbreaks in the U.S. associated with consumption of both fresh and dried specialty mushrooms have recently occurred. Dried wood ear mushrooms were...
Foodborne illness outbreaks in the U.S. associated with consumption of both fresh and dried specialty mushrooms have recently occurred. Dried wood ear mushrooms were implicated in a salmonellosis outbreak in 2020, while fresh enoki mushrooms were associated with two listeriosis outbreaks in 2020 and 2023. These specialty mushrooms are commercially available in both their fresh and dried states. Due to the short shelf life of mushrooms, dehydration is a common method used in both industry and by consumers to extend the shelf life and preserve quality. Therefore, the aim of this study was to evaluate the use of dehydration on the inactivation kinetics of both and on enoki and wood ear mushrooms. Fresh mushrooms were inoculated with four strain cocktails of either or and dried at ambient conditions for 10 min. Following drying of the inoculum, mushrooms were placed into food dehydrators preheated to 70, 80, or 90°C and treated for up to 24 h. At treatment intervals, mushrooms were removed from the dehydrators for pathogen enumeration. Inactivation kinetics for both pathogens were modeled using the Weibull, log-linear with tail, and log-linear with shoulder models. Pathogen reductions of >4 log CFU/g were achieved on both enoki and wood ear mushrooms during dehydration at 90°C after only 2-4 h. At 70 and 80°C, log reductions of >4 log CFU/g were observed on wood ear mushrooms after 4-8 h. On enoki mushrooms, a tailing effect was observed with residual populations (>2 log CFU/g) of and remaining even after 24 h of treatment at both 70 and 80°C. This study emphasizes the need for an individualized dehydration strategy for each mushroom type to ensure the effectiveness of dehydration as a process to reduce pathogen populations. Results of this study will aid in informing proper time and temperature combinations for dehydration of specialty mushrooms to ensure product safety.
PubMed: 38029214
DOI: 10.3389/fmicb.2023.1257053 -
Microorganisms Oct 2023() is ubiquitous in nature and known for its ability to contaminate foods during production processes. Near real-time monitoring of whole genome sequences from food and...
() is ubiquitous in nature and known for its ability to contaminate foods during production processes. Near real-time monitoring of whole genome sequences from food and human isolates, complemented with epidemiological data, has been used in the Netherlands since 2019 to increase the speed and success rate of source finding in the case of (active) clusters. Nine clusters with 4 to 19 human cases investigated between January 2019 and May 2023 are described. Fish production sites were most often linked to outbreaks of listeriosis (six clusters), though other types of food businesses can face similar problems, as the production processes and procedures determine risk. The results showed that low levels of in food samples can still be linked to disease. Therefore, the investigation of a cluster of cases and deployment of the precautionary principle helps to focus on safe food and to prevent further cases. Good practice of environmental monitoring within a food business allows early detection of potential issues with food safety and helps food businesses to take appropriate measures such as cleaning to prevent regrowth of and thus future outbreaks.
PubMed: 38004629
DOI: 10.3390/microorganisms11112617 -
Journal of Infection in Developing... Dec 2023There are no randomized controlled trials to inform the choice of using adjunctive dexamethasone (AD) against Listeria monocytogenes meningitis (LMM) and data from... (Review)
Review Meta-Analysis
Predisposing factor, impactful adjunctive treatment or unfavorable prognostic marker: a meta-analysis on the elusive role of dexamethasone for Listeria monocytogenes meningitis.
INTRODUCTION
There are no randomized controlled trials to inform the choice of using adjunctive dexamethasone (AD) against Listeria monocytogenes meningitis (LMM) and data from observational studies are pretty conflicting.
METHODOLOGY
We performed a rapid review of the literature with quantitative analysis. A pairwise random-effects meta-analysis was implemented, pooling unadjusted and adjusted data. The main outcome was mortality.
RESULTS
Across all included studies (five) informing the main analysis on raw mortality data, 199 patients received AD, as opposed to 382 who did not receive AD. All-cause mortality was slightly lower in patients undergoing AD, but not in a statistically significant manner: odds ratio 0.96, 95% confidence interval 0.42-2.19. The prediction interval was very wide (0.06-15.99), suggesting that in future studies the effect of AD might be either beneficial or harmful.
CONCLUSIONS
The role of AD for LMM still needs to be established being the current evidence inconclusive and heterogeneous.
Topics: Humans; Meningitis, Listeria; Prognosis; Dental Care; Causality; Dexamethasone
PubMed: 38252726
DOI: 10.3855/jidc.18804 -
Microorganisms Feb 2024() is a pathogen that is transmitted through contaminated food and causes the illness known as listeriosis. The virulence factor InlA plays a crucial role in the...
() is a pathogen that is transmitted through contaminated food and causes the illness known as listeriosis. The virulence factor InlA plays a crucial role in the invasion of into the human intestinal epithelium. In addition, InlA enhances the pathogenicity of host strains, and different strains of contain varying variations of InlA. Our study analyzed a total of 4393 published genomes from 511 sequence types (STs) of diverse origins. We identified 300 unique InlA protein sequence types (PSTs) and revealed 45 highly mutated amino acid sites. The leucine-rich repeat (LRR) region was found to be the most conserved among the InlA, while the protein A (PA) region experienced the highest mutation rate. Two new types of mutations were identified in the B-repeat region of InlA. Correspondence analysis (CA) was used to analyze correlations between the lineages or 10 most common sequence types (STs) and amino acid (aa) sites. ST8 was strongly correlated with site 192_F, 454_T. ST7 exhibited a strong correlation with site 51_A, 573_E, 648_S, and 664_A, and it was also associated with ST6 and site 544_N, 671_A, 738_B, 739_B, 740_B, and 774_Y. Additionally, a strong correlation between ST1 and site 142_S, 738_N, ST2 and site 2_K, 142_S, 738_N, as well as ST87 and site2_K, 738_N was demonstrated. Our findings contribute significantly to the understanding of the distribution, composition, and conservation of InlA in These findings also suggest a potential role of InlA in supporting molecular epidemiological tracing efforts.
PubMed: 38543536
DOI: 10.3390/microorganisms12030485 -
Journal of Food Protection Aug 2023In 2016, the U.S. Food and Drug Administration (FDA), the Centers for Disease Control and Prevention (CDC), and state partners investigated nine Listeria monocytogenes...
Investigation of a Multistate Outbreak of Listeria monocytogenes Infections Linked to Frozen Vegetables Produced at Individually Quick-Frozen Vegetable Manufacturing Facilities.
In 2016, the U.S. Food and Drug Administration (FDA), the Centers for Disease Control and Prevention (CDC), and state partners investigated nine Listeria monocytogenes infections linked to frozen vegetables. The investigation began with two environmental L. monocytogenes isolates recovered from Manufacturer A, primarily a processor of frozen onions, that were a match by whole genome sequencing (WGS) to eight clinical isolates and historical onion isolates with limited collection details. Epidemiologic information, product distribution, and laboratory evidence linked suspect food items, including products sourced from Manufacturer B, also a manufacturer of frozen vegetable/fruit products, with an additional illness. The environmental isolates were obtained during investigations at Manufacturers A and B. State and federal partners interviewed ill people, analyzed shopper card data, and collected household and retail samples. Nine ill persons between 2013 and 2016 were reported in four states. Of four ill people with information available, frozen vegetable consumption was reported by three, with shopper cards confirming purchases of Manufacturer B brands. Two identified outbreak strains of L. monocytogenes (Outbreak Strain 1 and Outbreak Strain 2) were a match to environmental isolates from Manufacturer A and/or isolates from frozen vegetables recovered from open and unopened product samples sourced from Manufacturer B; the investigation resulted in extensive voluntary recalls. The close genetic relationship between isolates helped investigators determine the source of the outbreak and take steps to protect public health. This is the first known multistate outbreak of listeriosis in the United States linked to frozen vegetables and highlights the significance of sampling and WGS analyses when there is limited epidemiologic information. Additionally, this investigation emphasizes the need for further research regarding food safety risks associated with frozen foods.
Topics: Humans; United States; Listeria monocytogenes; Vegetables; Foodborne Diseases; Food Microbiology; Listeriosis; Disease Outbreaks; Onions
PubMed: 37327999
DOI: 10.1016/j.jfp.2023.100117 -
Health Science Reports Feb 2024Recurring outbreaks in the United States is a growing public healthcare concern. Although no associated reported death, 17 were hospitalized out of the 18 reported...
Recurring outbreaks in the United States is a growing public healthcare concern. Although no associated reported death, 17 were hospitalized out of the 18 reported illnesses in the recent outbreak in 15 US states. The United States has experienced about 30 outbreaks in the last decade with 524 Listeriosis cases and 80 deaths. The identified origin were ice cream, leafy greens, mushroom, meat slice, dairy products like cheese, packaged salads, cooked chicken, hard-boiled egg, pork product, frozen vegetable, raw milk, packaged caramel apple, bean sprout and soya products. Although rare, may lead to serious illness (invasive listeriosis) or death. Listeriosis is critically harmful and medically complicated, especially in the pregnant, the old above 65 years and in the immunocompromised. It could cause premature birth, miscarriage or even neonatal death. Hospitalization is often necessary in the geriatric, being fatal at times. Among sp., is often human infection-associated. It is a gram-positive, non-sporulating, motile bacillus opportunistic pathogen. Food-borne listeriosis is often associated with frozen foods due to its ability to thrive at low temperatures. Hypervirulent strains of with an ability to infect the respiratory system (the lungs) was recently reported in the coronavirus disease-19 patients during the pandemic. seemed to have developed antimicrobial resistance to ciprofloxacin and meropenem, possibly acquired through the food chain. An early onset of listeriosis in the newborn is evident in the first 7 days postparturition. As the bacteria colonize the genitourinary tract, majority of such cases result from teratogenic transfer during vaginal delivery. Premature newborns, neonates born outside healthcare facilities and low-birth-weight babies were increasingly predisposed to an early onset of listeriosis. outbreaks were earlier reported in South Africa, Australia and Europe, with an unclear origin of the outbreaks. Social media updates about such outbreaks, the most likely food source, and measures to self-protect are suggested as preventive measures. The article deals on various such aspects related to listeriosis primarily originating from food, to ensure better public healthcare and human wellness.
PubMed: 38317674
DOI: 10.1002/hsr2.1863 -
Journal of Controlled Release :... Jun 2024Although various types of mRNA-based vaccines have been explored, the optimal conditions for induction of both humoral and cellular immunity remain rather unknown. In...
Although various types of mRNA-based vaccines have been explored, the optimal conditions for induction of both humoral and cellular immunity remain rather unknown. In this study, mRNA vaccines of nucleoside-modified mRNA in lipoplexes (LPXs) or lipid nanoparticles (LNPs) were evaluated after administration in mice through different routes, assessing mRNA delivery, tolerability and immunogenicity. In addition, we investigated whether mRNA vaccines could benefit from the inclusion of the adjuvant alpha-galactosylceramide (αGC), an invariant Natural Killer T (iNKT) cell ligand. Intramuscular (IM) vaccination with ovalbumin (OVA)-encoding mRNA encapsulated in LNPs adjuvanted with αGC showed the highest antibody- and CD8 T cell responses. Furthermore, we observed that addition of signal peptides and endocytic sorting signals of either LAMP1 or HLA-B7 in the OVA-encoding mRNA sequence further enhanced CD8 T cell activation although reducing the induction of IgG antibody responses. Moreover, mRNA LNPs with the ionizable lipidoid C12-200 exhibited higher pro-inflammatory- and reactogenic activity compared to mRNA LNPs with SM-102, correlating with increased T cell activation and antitumor potential. We also observed that αGC could further enhance the cellular immunity of clinically relevant mRNA LNP vaccines, thereby promoting therapeutic antitumor potential. Finally, a Listeria monocytogenes mRNA LNP vaccine supplemented with αGC showed synergistic protective effects against listeriosis, highlighting a key advantage of co-activating iNKT cells in antibacterial mRNA vaccines. Taken together, our study offers multiple insights for optimizing the design of mRNA vaccines for disease applications, such as cancer and intracellular bacterial infections.
Topics: Animals; Galactosylceramides; Mice, Inbred C57BL; Cancer Vaccines; Female; Nanoparticles; Ovalbumin; mRNA Vaccines; Adjuvants, Immunologic; CD8-Positive T-Lymphocytes; RNA, Messenger; Mice; Bacterial Vaccines; Neoplasms; Lipids; Liposomes
PubMed: 38697317
DOI: 10.1016/j.jconrel.2024.04.052