Listeria exploits IFITM3 to suppress antibacterial activity in phagocytes.

The type I interferon (IFN) signaling pathway has important functions in resistance to viral infection, with the downstream induction of interferon stimulated genes (ISG) protecting the host from virus entry, replication and spread. Listeria monocytogenes (Lm), a facultative intracellular foodborne pathogen, can exploit the type I IFN response as part of their pathogenic strategy, but the molecular mechanisms involved remain unclear. Here we show that type I IFN suppresses the antibacterial activity of phagocytes to promote systemic Lm infection. Mechanistically, type I IFN suppresses phagosome maturation and proteolysis of Lm virulence factors ActA and LLO, thereby promoting phagosome escape and cell-to-cell spread; the antiviral protein, IFN-induced transmembrane protein 3 (IFITM3), is required for this type I IFN-mediated alteration. Ifitm3-/- mice are resistant to systemic infection by Lm, displaying decreased bacterial spread in tissues, and increased immune cell recruitment and pro-inflammatory cytokine signaling. Together, our findings show how an antiviral mechanism in phagocytes can be exploited by bacterial pathogens, and implicate IFITM3 as a potential antimicrobial therapeutic target.

Labeling Acidic Compartments of Neutrophils with Cresyl Violet.

We introduce the acidotropic marker cresyl violet to stain acidic granules in live neutrophils. Cresyl violet is less phototoxic, more photostable, and more cost-effective than other commercially available acidotropic markers. Additionally, it does not photoconvert to fluorescent species of a different color, a limitation of other commonly used acidotropic markers. Staining can be readily detected by fluorescence microscopy or by flow cytometry, and can be used as a readout of degranulation in activated neutrophils.

Dynamic Podosome-Like Structures in Nascent Phagosomes Are Coordinated by Phosphoinositides.

Phagocytosis, the engulfment of particulate matter, requires the coordinated polymerization of F-actin; however, the nature and dynamics of the F-actin structures generated during the process are incompletely defined. Using super-resolution microscopy, we observed the formation of podosome-like structures during Fc receptor-mediated phagocytosis. Unlike conventional podosomes, these structures are short lived and vectorial, expanding radially from the sites where phagocytic targets are initially engaged. The expanding ring of podosome-like structures requires the localized formation of PtdIns(3,4,5)P3. Concomitantly, the initial podosome-like structures disappear from the center of the phagocytic cup, enabling membrane bending around the target. This coordinated disappearance is mediated by localized hydrolysis of PtdIns(4,5)P2 at the center of the cup. Interference reflection microscopy revealed that the podosome-like structures attach tightly to the target, facilitating the progressive engagement and activation of phagocytic receptors, creating a diffusion barrier and serving as support for the extension of exploratory lamellipodia that probe the target surface.

Cresyl violet: a superior fluorescent lysosomal marker.

We have characterized cresyl violet as a membrane-permeant fluorophore that localizes to lysosomes and acidic vacuoles of budding yeast, Drosophila, human, murine and canine cells. An acidotropic weak base, cresyl violet is shown to be virtually insensitive to physiological alkali and divalent cations. Because of its unique spectral properties, it can be used in combination with green, red and far-red fluorophores, is less susceptible to photobleaching than alternative acidotropic probes, and does not undergo photoconversion. At concentrations that yield bright labeling of acidic compartments, cresyl violet does not alter the organellar pH nor does it affect the buffering capacity. Its affordability, together with its chemical and spectral properties, make cresyl violet a superior lysosomal marker devoid of many of the negative characteristics associated with other lysosomal probes.

Diffusion Barriers, Mechanical Forces, and the Biophysics of Phagocytosis.

Phagocytes recognize and eliminate pathogens, alert other tissues of impending threats, and provide a link between innate and adaptive immunity. They also maintain tissue homeostasis, consuming dead cells without causing alarm. The receptor engagement, signal transduction, and cytoskeletal rearrangements underlying phagocytosis are paradigmatic of other immune responses and bear similarities to macropinocytosis and cell migration. We discuss how the glycocalyx restricts access to phagocytic receptors, the processes that enable receptor engagement and clustering, and the remodeling of the actin cytoskeleton that controls the mobility of membrane proteins and lipids and provides the mechanical force propelling the phagocyte membrane toward and around the phagocytic prey.

Meta-Analysis of Serum Insulin-Like Growth Factor 1 in Alzheimer's Disease.

Insulin-like growth factor 1 (IGF-1) serum levels have been reported to be altered in Alzheimer's disease patients, and it was suggested that the changes in IGF-1 serum level may play a role in disease pathology and progression. However, this notion remained controversial due to conflicting findings. We conducted a meta-analysis to determine the relationship between IGF-1 serum levels and Alzheimer's disease. We searched the databases PUBMED, Ovid SP, and Cochrane library for relevant studies. The primary data analyzed was serum IGF-1 from Alzheimer's disease subjects and controls. Pooled weighted mean difference using a random effects model was used to determine the relationship between serum levels and disease state. Nine studies were included in the meta-analysis compromising a total of 1639 subjects. The pooled weighted mean difference was -2.27ng/ml (95% CI: [-22.221, 17.66]) with a P value of 0.82. Thus our finding did not show clear relationship between low IGF-1 and Alzheimer's disease subjects. We did not find evidence of publication bias by analyzing a funnel plot as well as Egger's and Begg's tests. While eight out of the nine studies included in this meta-analysis detected a statistically significant increase or decrease in serum levels of IGF-1 in Alzheimer's disease subjects, the analysis as a whole did not show a significant trend in either direction. Thus, IGF-1 level is likely a critical personalized factor. A large database of clinical trials is required for better understanding the relationship between IGF-1 levels and Alzheimer's disease.