Delay of phagosome maturation by a mycobacterial lipid is reversed by nitric oxide.

Mycobacterium tuberculosis is a facultative intracellular pathogen that inhibits phagosome maturation in macrophages thereby securing survival and growth. Mycobacteria reside in an early endocytic compartment of near-neutral pH where they upregulate production of complex glycolipids such as trehalose dimycolate. Here, we report that trehalose dimycolate coated onto beads increased the bead retention in early phagosomes, i.e. at a similar stage as viable mycobacteria. Thus, a single mycobacterial lipid sufficed to divert phagosome maturation and likely contributes to mycobacterial survival in macrophages. Previous studies showed that activated macrophages promote maturation of mycobacterial phagosomes and eliminate mycobacteria through bactericidal effectors including nitric oxide generated by inducible nitric-oxide synthase. We show that deceleration of bead phagosome maturation by trehalose dimycolate was abolished in immune-activated wild type, but not in activated nitric-oxide synthase-deficient macrophages, nor when hydroxyl groups of trehalose dimycolate were chemically modified by reactive nitrogen intermediates. Thus, specific host defence effectors of activated macrophages directly target a specific virulence function of mycobacteria

Correction of the Iron Overload Defect in β-2-Microglobulin Knockout Mice by Lactoferrin Abolishes Their Increased Susceptibility to Tuberculosis

As a resident of early endosomal phagosomes, Mycobacterium tuberculosis is connected to the iron uptake system of the host macrophage. β-2-microglobulin (β2m) knockout (KO) mice are more susceptible to tuberculosis than wild-type mice, which is generally taken as a proof for the role of major histocompatibility complex class I (MHC-I)–restricted CD8 T cells in protection against M. tuberculosis. However, β2m associates with a number of MHC-I–like proteins, including HFE. This protein regulates transferrin receptor mediated iron uptake and mutations in its gene cause hereditary iron overload (hemochromatosis). Accordingly, β2m-deficient mice suffer from tissue iron overload. Here, we show that modulating the extracellular iron pool in β2m–KO mice by lactoferrin treatment significantly reduces the burden of M. tuberculosis to numbers comparable to those observed in MHC class I–KO mice. In parallel, the generation of nitric oxide impaired in β2m–KO mice was rescued. Conversely, iron overload in the immunocompetent host exacerbated disease. Consistent with this, iron deprivation in infected resting macrophages was detrimental for intracellular mycobacteria. Our data establish: (a) defective iron metabolism explains the increased susceptibility of β2m-KO mice over MHC-I–KO mice, and (b) iron overload represents an exacerbating cofactor for tuberculosis

The IFN-inducible Golgi- and endoplasmic reticulum- associated 47-kDa GTPase IIGP is transiently expressed during listeriosis.

Members of the 47-kDa GTPase family are implicated in an IFN-gamma-induced, as yet unclear, mechanism that confers innate resistance against infection with intracellular pathogens. Overt immunological parameters are apparently uncompromised in mice deficient for individual members and the prototype of this family, IGTP, localizes to the endoplasmic reticulum. This suggests that these GTPases are involved in intracellular defense. We analyzed the expression of the 47-kDa GTPase cognate, IIGP, in splenic sections from mice infected with the intracellular pathogen Listeria monocytogenes by immunohistochemistry. An early transient IIGP induction was observed revealing the IFN-gamma responsiveness of cellular subcompartments within the spleen in early listeriosis. Marginal metallophilic macrophages and endothelial cells within the red and white pulp strongly expressed IIGP, while other splenocytes remained negative. In vitro analyses show that both type I and type II IFNs are prime stimuli for IIGP induction in various cells, including L. monocytogenes-infected or LPS-stimulated macrophages, endothelial cells, and activated T cells. Contrary to the subcellular localization of IGTP, IIGP was predominantly associated with the Golgi apparatus and also localizes to the endoplasmic reticulum. We conclude that IIGP exerts a distinct role in IFN-induced intracellular membrane trafficking or processing

IL-4 and T cells are required for the generation of IgG1 isotype antibodies against cardiolipin.

Infection with Mycobacterium tuberculosis induces Abs against a vast array of mycobacterial lipids and glycolipids. One of the most prominent lipid Ags recognized is cardiolipin (CL). The kinetics of the generation of anti-CL Abs during infection reveals that IgM titers to CL increase over time. Interestingly, at day 30 postinfection CL-specific IgG1 appears, an isotype usually dependent on T cell help. Using an immunization schedule with CL/anti-CL Ab complexes, which induces antiphospholipid syndrome in mice, we show that the generation of IgG1 to CL requires IL-4 and that optimal production is T cell dependent. IgG1 production to CL was impaired in nude (nu/nu) mice devoid in conventional T cells, but was not affected in mice deficient for either alphabeta TCR(+), gammadelta TCR(+), CD4(+), CD8(+), or NK1.1(+) T cells. We conclude that IgG1 production to CL depends on T cell help and IL-4, which can be provided by different T cell populations. This is the first report that IL-4 is indispensable for the induction of IgG1 Abs to lipid Ags

Familienpolitik: Förderung von Familien - nach welchem Konzept?

Familienpolitik hat Konjunktur und ist ins Zentrum der politischen Agenda gerückt, weil sie eng mit Demographie- und Wachstumspolitik verbunden ist. Über die Notwendigkeit einer wirksamen Familienpolitik herrscht Einigkeit, aber welches Konzept ist das richtige? Christa Stewens, Bayerische Staatsministerin für Arbeit und Sozialordnung, Familie und Frauen, unterstreicht, dass Familienpolitik "ganzheitlich und nachhaltig angelegt sein" muss. Auch Malte Ristau, Bundesministerium für Familie, Senioren, Frauen und Jugend, verweist darauf, dass eine nur auf monetäre Hilfe angelegte Familienpolitik ein zu enger Ansatz sei. Reiner Klingholz, Berlin-Institut für Bevölkerung und Entwicklung, zeigt, dass interessanterweise nicht die Länder, in denen ein traditionelles Frauen- und Familienbild vorherrscht, sondern jene, in denen die Gleichstellung der Geschlechter am weitesten fortgeschritten ist, die höchste Fertilität aufweisen. Bertram Wiest und Stefan Schaible, Roland Berger Strategy Consultants, betonen das hohe Finanzvolumen, das in Deutschland vom Staat für Leistungen für Familien ausgegeben wird: "Es mangelt also nicht an Geld, sondern am zielgerichteten Mitteleinsatz". Für Michael Steiner, Michael Böhmer und Christian Böllhoff, Prognos AG, Basel, muss sich die Familienpolitik an das neue Familienbild anpassen und sowohl Gestaltungsfreiräume für Familie einrichten, als auch Chancengleichheit gewährleisten. Auch für Christel Humme, familienpolitische Sprecherin der SPD-Bundestagsfraktion, kommt es vor allem auf die richtige Mischung zwischen "Infrastruktur, Zeit und Geld für Familien" an.Familienpolitik, Konjunktur, Demographie, Wachstumspolitik, Kinder, Familie, Deutschland

Basic study on the evaluation of thermoplastic polymers as hot-melt adhesives for mixed-substrate joining

A selection of 22 low-melting polymers was thermally and rheologically evaluated to be used as hot-melt adhesives in mixed-substrate joining samples. The choice of polymers was based on the published melting point. It was required to include a broad variety of different polymers backbones to study the influence of the different polymers comprehensively. A tool-box of widely applicable tests was developed to judge if a thermoplastic polymer is suitable for a hot-melt adhesive application. Melting temperature (onset, peak and offset temperature) and melting enthalpy were determined using standardized methods. Rheological methods were used to characterize the shear rate dependence and the flow behavior at the application temperature. The wetting behavior of the polymers was evaluated with contact angle measurements. The adhesive strength of the most promising candidates was analyzed using the Lumi Frac-adhesion method including the failure pattern

Mycobacterium tuberculosis Complex Lineage 3 as Causative Agent of Pulmonary Tuberculosis, Eastern Sudan1.

Pathogen-based factors associated with tuberculosis (TB) in eastern Sudan are not well defined. We investigated genetic diversity, drug resistance, and possible transmission clusters of Mycobacterium tuberculosis complex (MTBC) strains by using a genomic epidemiology approach. We collected 383 sputum specimens at 3 hospitals in 2014 and 2016 from patients with symptoms suggestive of TB; of these, 171 grew MTBC strains. Whole-genome sequencing could be performed on 166 MTBC strains; phylogenetic classification revealed that most (73.4%; n = 122) belonged to lineage 3 (L3). Genome-based cluster analysis showed that 76 strains (45.9%) were grouped into 29 molecular clusters, comprising 2-8 strains/patients. Of the strains investigated, 9.0% (15/166) were multidrug resistant (MDR); 10 MDR MTBC strains were linked to 1 large MDR transmission network. Our findings indicate that L3 strains are the main causative agent of TB in eastern Sudan; MDR TB is caused mainly by transmission of MDR L3 strains

Peripheral sensitisation of nociceptors via G-proteindependent potentiation of mechanotransduction currents

Mechanical stimuli impinging on the skin are converted into electrical signals by mechanically gated ion channels located at the peripheral nerve endings of dorsal root ganglion (DRG) neurons. Under inflammatory conditions sensory neurons are commonly sensitised to mechanical stimuli; a putative mechanism that may contribute to such sensitisation of sensory neurons is enhanced responsiveness of mechanotransduction ion channels. Here we show that the algogens UTP and ATP potentiate mechanosensitive RA currents in peptidergic nociceptive DRG neurons and reduce thresholds for mechanically induced action potential firing in these neurones. Pharmacological characterisation suggests that this effect is mediated by the Gq-coupled P2Y2 nucleotide receptor. Moreover, using the in vitro skin nerve technique, we show that UTP also increases action potential firing rates in response to mechanical stimuli in a subpopulation of skin C-fibre nociceptors. Together our findings suggest that UTP sensitises a subpopulation of cutaneous C-fibre nociceptors via a previously undescribed G-protein-dependent potentiation of mechanically activated RA-type currents