Iron-depleting nutritional immunity controls extracellular bacterial replication in Legionella pneumophila infections.

The accidental human pathogen Legionella pneumophila (Lp) is the etiological agent for a severe atypical pneumonia known as Legionnaires' disease. In human infections and animal models of disease alveolar macrophages are the primary cellular niche that supports bacterial replication within a unique intracellular membrane-bound organelle. The Dot/Icm apparatus-a type IV secretion system that translocates ~300 bacterial proteins within the cytosol of the infected cell-is a central virulence factor required for intracellular growth. Mutant strains lacking functional Dot/Icm apparatus are transported to and degraded within the lysosomes of infected macrophages. The early foundational work from Dr. Horwitz's group unequivocally established that Legionella does not replicate extracellularly during infection-a phenomenon well supported by experimental evidence for four decades. Our data challenges this paradigm by demonstrating that macrophages and monocytes provide the necessary nutrients and support robust Legionella extracellular replication. We show that the previously reported lack of Lp extracellular replication is not a bacteria intrinsic feature but rather a result of robust restriction by serum-derived nutritional immunity factors. Specifically, the host iron-sequestering protein Transferrin is identified here as a critical suppressor of Lp extracellular replication in an iron-dependent manner. In iron-overload conditions or in the absence of Transferrin, Lp bypasses growth restriction by IFNγ-primed macrophages though extracellular replication. It is well established that certain risk factors associated with development of Legionnaires' disease, such as smoking, produce a chronic pulmonary environment of iron-overload. Our work indicates that iron-overload could be an important determinant of severe infection by allowing Lp to overcome nutritional immunity and replicate extracellularly, which in turn would circumvent intracellular cell intrinsic host defenses. Thus, we provide evidence for nutritional immunity as a key underappreciated host defense mechanism in Legionella pathogenesis.

The intracellular growth of the vacuolar pathogen Legionella pneumophila is dependent on the acyl chain composition of host membranes.

Legionella pneumophila is an accidental human bacterial pathogen that infects and replicates within alveolar macrophages causing a severe atypical pneumonia known as Legionnaires' disease. As a prototypical vacuolar pathogen L. pneumophila establishes a unique endoplasmic reticulum (ER)-derived organelle within which bacterial replication takes place. Bacteria-derived proteins are deposited in the host cytosol and in the lumen of the pathogen-occupied vacuole via a type IVb (T4bSS) and a type II (T2SS) secretion system respectively. These secretion system effector proteins manipulate multiple host functions to facilitate intracellular survival of the bacteria. Subversion of host membrane glycerophospholipids (GPLs) by the internalized bacteria via distinct mechanisms feature prominently in trafficking and biogenesis of the Legionella -containing vacuole (LCV). Conventional GPLs composed of a glycerol backbone linked to a polar headgroup and esterified with two fatty acids constitute the bulk of membrane lipids in eukaryotic cells. The acyl chain composition of GPLs dictates phase separation of the lipid bilayer and therefore determines the physiochemical properties of biological membranes - such as membrane disorder, fluidity and permeability. In mammalian cells, fatty acids esterified in membrane GPLs are sourced endogenously from de novo synthesis or via internalization from the exogenous pool of lipids present in serum and other interstitial fluids. Here, we exploited the preferential utilization of exogenous fatty acids for GPL synthesis by macrophages to reprogram the acyl chain composition of host membranes and investigated its impact on LCV homeostasis and L. pneumophila intracellular replication. Using saturated fatty acids as well as cis - and trans - isomers of monounsaturated fatty acids we discovered that under conditions promoting lipid packing and membrane rigidification L. pneumophila intracellular replication was significantly reduced. Palmitoleic acid - a C16:1 monounsaturated fatty acid - that promotes membrane disorder when enriched in GPLs significantly increased bacterial replication within human and murine macrophages but not in axenic growth assays. Lipidome analysis of infected macrophages showed that treatment with exogenous palmitoleic acid resulted in membrane acyl chain reprogramming in a manner that promotes membrane disorder and live-cell imaging revealed that the consequences of increasing membrane disorder impinge on several LCV homeostasis parameters. Collectively, we provide experimental evidence that L. pneumophila replication within its intracellular niche is a function of the lipid bilayer disorder and hydrophobic thickness.

Cellular cholesterol licenses Legionella pneumophila intracellular replication in macrophages.

Host membranes are inherently critical for niche homeostasis of vacuolar pathogens. Thus, intracellular bacteria frequently encode the capacity to regulate host lipogenesis as well as to modulate the lipid composition of host membranes. One membrane component that is often subverted by vacuolar bacteria is cholesterol - an abundant lipid that mammalian cells produce de novo at the endoplasmic reticulum (ER) or acquire exogenously from serum-derived lipoprotein carriers. Legionella pneumophila is an accidental human bacterial pathogen that infects and replicates within alveolar macrophages causing a severe atypical pneumonia known as Legionnaires' disease. From within a unique ER-derived vacuole L. pneumophila promotes host lipogenesis and experimental evidence indicates that cholesterol production might be one facet of this response. Here we investigated the link between cellular cholesterol and L. pneumophila intracellular replication and discovered that disruption of cholesterol biosynthesis or cholesterol trafficking lowered bacterial replication in infected cells. These growth defects were rescued by addition of exogenous cholesterol. Conversely, bacterial growth within cholesterol-leaden macrophages was enhanced. Importantly, the growth benefit of cholesterol was observed strictly in cellular infections and L. pneumophila growth kinetics in axenic cultures did not change in the presence of cholesterol. Microscopy analyses indicate that cholesterol regulates a step in L. pneumophila intracellular lifecycle that occurs after bacteria begin to replicate within an established intracellular niche. Collectively, we provide experimental evidence that cellular cholesterol promotes L. pneumophila replication within a membrane bound organelle in infected macrophages.

Coarse graining in time with the functional renormalization group: Relaxation in Brownian motion.

We apply the functional renormalization group (fRG) to study relaxation in a stochastic process governed by an overdamped Langevin equation with one degree of freedom, exploiting the connection with supersymmetric quantum mechanics in imaginary time. After reviewing the functional integral formulation of the system and its underlying symmetries, including the resulting Ward-Takahashi identities for arbitrary initial conditions, we compute the effective action Γ from the fRG, approximated in terms of the leading and subleading terms in the gradient expansion: the local potential approximation and wave-function renormalization, respectively. This is achieved by coarse graining the thermal fluctuations in time resulting in, e.g., an effective potential incorporating fluctuations at all timescales. We then use the resulting effective equations of motion to describe the decay of the covariance and the relaxation of the average position and variance toward their equilibrium values at different temperatures. We use as examples a simple polynomial potential, an unequal Lennard-Jones type potential, and a more complex potential with multiple trapping wells and barriers. We find that these are all handled well, with the accuracy of the approximations improving as the relaxation's spectral representation shifts to lower eigenvalues, in line with expectations about the validity of the gradient expansion. The spectral representation's range also correlates with temperature, leading to the conclusion that the gradient expansion works better for higher temperatures than lower ones. This paper demonstrates the ability of the fRG to expedite the computation of statistical objects in otherwise long-timescale simulations, acting as a first step to more complicated systems.

Recent Advances in the Pursuit of an Effective Acinetobacter baumannii Vaccine.

Acinetobacter baumannii has been a major cause of nosocomial infections for decades. The absence of an available vaccine coupled with emerging multidrug resistance has prevented the medical community from effectively controlling this human pathogen. Furthermore, the ongoing pandemic caused by SARS-CoV-2 has increased the risk of hospitalized patients developing ventilator-associated pneumonia caused by bacterial opportunists including A. baumannii. The shortage of antibiotics in the development pipeline prompted the World Health Organization to designate A. baumannii a top priority for the development of new medical countermeasures, such as a vaccine. There are a number of important considerations associated with the development of an A. baumannii vaccine, including strain characteristics, diverse disease manifestations, and target population. In the past decade, research efforts have revealed a number of promising new immunization strategies that could culminate in a safe and protective vaccine against A. baumannii. In this review, we highlight the recent progress in the development of A. baumannii vaccines, discuss potential challenges, and propose future directions to achieve an effective intervention against this human pathogen.

Urban ministry workers' positive experiences of interpersonal and religious support during crisis.

Research on faith-based urban workers is limited despite the chronic and traumatic exposure inherent in their work. This study details the perception of positive interpersonal relationships during a time of trauma or crisis as described in semistructured 2- to 3-hour interviews with 13 faith-based urban workers in Los Angeles, California. Using strategies consistent with Consensual Qualitative Research, categories and subcategories defining positive interpersonal relationships were identified. Resulting categories suggested that there are specific characteristics, products, and types of relationships that urban workers experience as important during the time of trauma or crisis. Positive experiences were often religious in nature and included feeling supported, feeling connected, relationship growth, sharing and listening, authenticity, and feeling as through relationships facilitated personal growth or coping. The findings highlight participants' need for both practical support and relational support which reflects and enhances their spiritual commitment.

Barriers to recovery in communities exposed to disasters: Sri Lankan voices speak.

Disasters experienced by a community place all members at risk for physical and psychological harm. While natural resilience may help many to recover, there may be barriers that hinder the recovery process. This qualitative study was conducted to examine barriers to recovery in a community impacted by both war and the tsunami. A group of 43 ethnically diverse Sri Lankans (F = 63%) participated in six focus groups and provided their perspectives on barriers they perceived to impede their recovery from traumatic events. Grounded-theory-based data analysis revealed culture-general and culture-specific socio-economic, environmental, sociocultural, and individual barriers that participants identified as impeding their recovery. Interventions and health policies targeting these groups could focus on helping communities to overcome these barriers as a means of facilitating recovery in these beleaguered communities.