Real-time imaging of Leishmania mexicana-infected early phagosomes: a study using primary macrophages generated from green fluorescent protein-Rab5 transgenic mice

The small GTPase Rab5 is a key regulator of endosome/phagosome maturation and in intravesicular infections marks a phagosome stage at which decisions over pathogen replication or destruction are integrated. It is currently unclear whether Leishmania-infected phagosomes uniformly pass through a Rab5+ stage on their intracellular path to compartments with late endosomal/early lysosomal characteristics. Differences in routes and final compartments could have consequences for accessibility to antileishmanial drugs. Here, we generated a unique gfp-rab5 transgenic mouse model to visualize Rab5 recruitment to early parasite-containing phagosomes in primary host cells. Using real-time fluorescence imaging of phagosomes carrying Leishmania mexicana, we determined that parasite-infested phagosomes follow a uniform sequence of transient Rab5 recruitment. Residence in Rab5+ compartments was much shorter compared with phagosomes harboring latex beads. Furthermore, a comparative analysis of parasite life-cycle stages and mutants deficient in lpg1, the gene encoding the enzyme required for synthesis of the dominant surface lipophosphoglycan, indicated that parasite surface ligands and host cell receptors modulate pathogen residence times in Rab5+ phagosomes, but, as far as tested, had no significant effect on intracellular L. mexicana survival or replication.—Lippuner, C., Paape, D., Paterou, A., Brand, J., Richardson, M., Smith, A. J., Hoffmann, K., Brinkmann, V., Blackburn, C., Aebischer, T. Real-time imaging of Leishmania mexicana-infected early phagosomes: a study using primary macrophages generated from green fluorescent protein-Rab5 transgenic mice

Stable water isotopes and accumulation rates in the Union Glacier region, Ellsworth Mountains, West Antarctica, over the last 35 years

ntarctica is well known to be highly susceptible to atmospheric and oceanic warming. However, due to the lack of long-term and in situ meteorological observations, little is known about the magnitude of the warming and the meteorological conditions in the intersection region between the Antarctic Peninsula (AP), the West Antarctic Ice Sheet (WAIS) and the East Antarctic Ice Sheet (EAIS). Here we present new stable water isotope data (δ18O, δD, d excess) and accumulation rates from firn cores in the Union Glacier (UG) region, located in the Ellsworth Mountains at the northern edge of the WAIS. The firn core stable oxygen isotopes and the d excess exhibit no statistically significant trend for the period 1980–2014, suggesting that regional changes in near-surface air temperature and moisture source variability have been small during the last 35 years. Backward trajectory modelling revealed the Weddell Sea sector, Coats Land and Dronning Maud Land (DML) to be the main moisture source regions for the study site throughout the year. We found that mean annual δ18O (δD) values in the UG region are negatively correlated with sea ice concentrations (SICs) in the northern Weddell Sea but not influenced by large-scale modes of climate variability such as the Southern Annular Mode (SAM) and the El Niño–Southern Oscillation (ENSO). Only mean annual d-excess values show a weak positive correlation with the SAM. On average annual snow accumulation in the UG region amounts to 0.245 m w.e. a−1 in 1980–2014 and has slightly decreased during this period. It is only weakly related to sea ice conditions in the Weddell Sea sector and not correlated with SAM and ENSO. We conclude that neither the rapid warming nor the large increases in snow accumulation observed on the AP and in West Antarctica during the last decades have extended inland to the Ellsworth Mountains. Hence, the UG region, although located at the northern edge of the WAIS and relatively close to the AP, exhibits rather stable climate characteristics similar to those observed in East Antarctica

Deciphering stable water isotope records of firn cores from a strongly maritime, high-accumulation site on the Antarctic Peninsula

Stable water isotope records of six firn cores retrieved from two adjacent plateaus on the northern Antarctic Peninsula between 2014 and 2016 are presented and investigated for their connections with firn-core glacio-chemical data, meteorological records and modelling results. Average annual accumulation rates of 2500 kg m−2 a−1 largely reduce the modification of isotopic signals in the snowpack by post-depositional processes, allowing excellent signal preservation in space and time. Comparison of firn-core and ECHAM6-wiso modelled δ18O and d-excess records reveals a large agreement on annual and sub-annual scales, suggesting firn-core stable water isotopes to be representative of specific synoptic situations. The six firn cores exhibit highly similar isotopic patterns in the overlapping period (2013), which seem to be related to temporal changes in moisture sources rather than local near-surface air temperatures. Backward trajectories calculated with the HYSPLIT model suggest that prominent δ18O minima in 2013 associated with elevated sea salt concentrations are related to long-range moisture transport dominated by westerly winds during positive SAM phases. In contrast, a broad δ18O maximum in the same year accompanied by increased concentrations of black carbon and mineral dust corresponds to the advection of more locally derived moisture with northerly flow components (South America) when the SAM is negative

SARS-CoV-2 variant Alpha has a spike-dependent replication advantage over the ancestral B.1 strain in human cells with low ACE2 expression

Epidemiological data demonstrate that Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) variants of concern (VOCs) Alpha and Delta are more transmissible, infectious, and pathogenic than previous variants. Phenotypic properties of VOC remain understudied. Here, we provide an extensive functional study of VOC Alpha replication and cell entry phenotypes assisted by reverse genetics, mutational mapping of spike in lentiviral pseudotypes, viral and cellular gene expression studies, and infectivity stability assays in an enhanced range of cell and epithelial culture models. In almost all models, VOC Alpha spread less or equally efficiently as ancestral (B.1) SARS-CoV-2. B.1. and VOC Alpha shared similar susceptibility to serum neutralization. Despite increased relative abundance of specific sgRNAs in the context of VOC Alpha infection, immune gene expression in infected cells did not differ between VOC Alpha and B.1. However, inferior spreading and entry efficiencies of VOC Alpha corresponded to lower abundance of proteolytically cleaved spike products presumably linked to the T716I mutation. In addition, we identified a bronchial cell line, NCI-H1299, which supported 24-fold increased growth of VOC Alpha and is to our knowledge the only cell line to recapitulate the fitness advantage of VOC Alpha compared to B.1. Interestingly, also VOC Delta showed a strong (595-fold) fitness advantage over B.1 in these cells. Comparative analysis of chimeric viruses expressing VOC Alpha spike in the backbone of B.1, and vice versa, showed that the specific replication phenotype of VOC Alpha in NCI-H1299 cells is largely determined by its spike protein. Despite undetectable ACE2 protein expression in NCI-H1299 cells, CRISPR/Cas9 knock-out and antibody-mediated blocking experiments revealed that multicycle spread of B.1 and VOC Alpha required ACE2 expression. Interestingly, entry of VOC Alpha, as opposed to B.1 virions, was largely unaffected by treatment with exogenous trypsin or saliva prior to infection, suggesting enhanced resistance of VOC Alpha spike to premature proteolytic cleavage in the extracellular environment of the human respiratory tract. This property may result in delayed degradation of VOC Alpha particle infectivity in conditions typical of mucosal fluids of the upper respiratory tract that may be recapitulated in NCI-H1299 cells closer than in highly ACE2-expressing cell lines and models. Our study highlights the importance of cell model evaluation and comparison for in-depth characterization of virus variant-specific phenotypes and uncovers a fine-tuned interrelationship between VOC Alpha- and host cell-specific determinants that may underlie the increased and prolonged virus shedding detected in patients infected with VOC Alpha

The chemokine receptor CXCR5 is pivotal for ectopic mucosa-associated lymphoid tissue neogenesis in chronic Helicobacter pylori-induced inflammation

Ectopic lymphoid follicles are a key feature of chronic inflammatory autoimmune and infectious diseases, such as rheumatoid arthritis, Sjögren's syndrome, and Helicobacter pylori-induced gastritis. Homeostatic chemokines are considered to be involved in the formation of such tertiary lymphoid tissue. High expression of CXCL13 and its receptor, CXCR5, has been associated with the formation of ectopic lymphoid follicles in chronic infectious diseases. Here, we defined the role of CXCR5 in the development of mucosal tertiary lymphoid tissue and gastric inflammation in a mouse model of chronic H. pylori infection. CXCR5-deficient mice failed to develop organized gastric lymphoid follicles despite similar bacterial colonization density as infected wild-type mice. CXCR5 deficiency altered Th17 responses but not Th1-type cellular immune responses to H. pylori infection. Furthermore, CXCR5-deficient mice exhibited lower H. pylori-specific serum IgG and IgA levels and an overall decrease in chronic gastric immune responses. In conclusion, the development of mucosal tertiary ectopic follicles during chronic H. pylori infection is strongly dependent on the CXCL13/CXCR5 signaling axis, and lack of de novo lymphoid tissue formation attenuates chronic immune responses

Dissecting the Shared Genetic Architecture of Suicide Attempt, Psychiatric Disorders, and Known Risk Factors

Background Suicide is a leading cause of death worldwide, and nonfatal suicide attempts, which occur far more frequently, are a major source of disability and social and economic burden. Both have substantial genetic etiology, which is partially shared and partially distinct from that of related psychiatric disorders. Methods We conducted a genome-wide association study (GWAS) of 29,782 suicide attempt (SA) cases and 519,961 controls in the International Suicide Genetics Consortium (ISGC). The GWAS of SA was conditioned on psychiatric disorders using GWAS summary statistics via multitrait-based conditional and joint analysis, to remove genetic effects on SA mediated by psychiatric disorders. We investigated the shared and divergent genetic architectures of SA, psychiatric disorders, and other known risk factors. Results Two loci reached genome-wide significance for SA: the major histocompatibility complex and an intergenic locus on chromosome 7, the latter of which remained associated with SA after conditioning on psychiatric disorders and replicated in an independent cohort from the Million Veteran Program. This locus has been implicated in risk-taking behavior, smoking, and insomnia. SA showed strong genetic correlation with psychiatric disorders, particularly major depression, and also with smoking, pain, risk-taking behavior, sleep disturbances, lower educational attainment, reproductive traits, lower socioeconomic status, and poorer general health. After conditioning on psychiatric disorders, the genetic correlations between SA and psychiatric disorders decreased, whereas those with nonpsychiatric traits remained largely unchanged. Conclusions Our results identify a risk locus that contributes more strongly to SA than other phenotypes and suggest a shared underlying biology between SA and known risk factors that is not mediated by psychiatric disorders.Peer reviewe

Classical Human Leukocyte Antigen Alleles and C4 Haplotypes Are Not Significantly Associated With Depression

Background The prevalence of depression is higher in individuals with autoimmune diseases, but the mechanisms underlying the observed comorbidities are unknown. Shared genetic etiology is a plausible explanation for the overlap, and in this study we tested whether genetic variation in the major histocompatibility complex (MHC), which is associated with risk for autoimmune diseases, is also associated with risk for depression. Methods We fine-mapped the classical MHC (chr6: 29.6–33.1 Mb), imputing 216 human leukocyte antigen (HLA) alleles and 4 complement component 4 (C4) haplotypes in studies from the Psychiatric Genomics Consortium Major Depressive Disorder Working Group and the UK Biobank. The total sample size was 45,149 depression cases and 86,698 controls. We tested for association between depression status and imputed MHC variants, applying both a region-wide significance threshold (3.9 × 10−6) and a candidate threshold (1.6 × 10−4). Results No HLA alleles or C4 haplotypes were associated with depression at the region-wide threshold. HLA-B*08:01 was associated with modest protection for depression at the candidate threshold for testing in HLA genes in the meta-analysis (odds ratio = 0.98, 95 confidence interval = 0.97–0.99). Conclusions We found no evidence that an increased risk for depression was conferred by HLA alleles, which play a major role in the genetic susceptibility to autoimmune diseases, or C4 haplotypes, which are strongly associated with schizophrenia. These results suggest that any HLA or C4 variants associated with depression either are rare or have very modest effect sizes

Short-Term Meteorological and Environmental Signals Recorded in a Firn Core from a High-Accumulation Site on Plateau Laclavere, Antarctic Peninsula

High-accumulation sites are crucial for understanding the patterns and mechanisms of climate and environmental change in Antarctica since they allow gaining high-resolution proxy records from firn and ice. Here, we present new glacio- and isotope-geochemical data at sub-annual resolution from a firn core retrieved from an ice cap on Plateau Laclavere (LCL), northern Antarctic Peninsula, covering the period 2012–2015. The signals of two volcanic eruptions and two forest fire events in South America could be identified in the non-sea-salt sulphur and black carbon records, respectively. Mean annual snow accumulation on LCL amounts to 2500 kg m−2 a−1 and exhibits low inter-annual variability. Time series of δ18O, δD and d excess show no seasonal cyclicity, which may result from (1) a reduced annual temperature amplitude due to the maritime climate and (2) post-depositional processes. The firn core stratigraphy indicates strong surface melt on LCL during austral summers 2013 and 2015, likely related to large-scale warm-air advection from lower latitudes and temporal variations in sea ice extent in the Bellingshausen-Amundsen Sea. The LCL ice cap is a highly valuable natural archive since it captures regional meteorological and environmental signals as well as their connection to the South American continent.Peer Reviewe