Spatial Separation of HLA-DM/HLA-DR Interactions within MIIC and Phagosome-Induced Immune Escape

SummaryMajor Histocompatibility Complex (MHC) class II molecules, including Human Leukocyte Antigen (HLA)-DR, present peptide fragments from proteins degraded in the endocytic pathway. HLA-DR is targeted to late-endocytic structures named MHC class II-containing Compartments (MIIC), where it interacts with HLA-DM. This chaperone stabilizes HLA-DR during peptide exchange and is critical for successful peptide loading. To follow this process in living cells, we have generated cells containing HLA-DR3/Cyan Fluorescent Protein (CFP), HLA-DM/Yellow Fluorescent Protein (YFP), and invariant chain. HLA-DR/DM interactions were observed by Fluorescence Resonance Energy Transfer (FRET). These interactions were pH insensitive, yet occurred only in internal structures and not at the limiting membrane of MIIC. In a cellular model of infection, phagosomes formed a limiting membrane surrounding internalized Salmonella. HLA-DR and HLA-DM did not interact in Salmonella-induced vacuoles, and HLA-DR was not loaded with antigens. The absence of HLA-DR/DM interactions at the limiting membrane prevents local loading of MHC class II molecules in phagosomes. This may allow these bacteria to successfully evade the immune system

Een nieuwe en handzame lysimeter: eerste stap naar een nationaal netwerk voor de werkelijke verdamping?

Vrijwel overal op aarde verdampt meer dan de helft van het neerslagwater, ook in Nederland. Toch wordt deze grote verliespost in ons land slechts sporadisch gemeten. Door inspanningen van kennisinstituten, bedrijven en overheid is daarom een lysimeter ontwikkeld. Metingen in 2014 en 2015 op twee locaties vertonen opvallende verschillen, maar ook grote overeenkomsten, met eddy-correlatiemetingen. Schattingen van de verdamping via satellietdata zijn hoger dan de metingen. Tijd daarom, om metingen in het veld te combineren met modellen en waarnemingen vanuit de ruimt

Tissue clonality of dendritic cell subsets and emergency DCpoiesis revealed by multicolor fate mapping of DC progenitors

Conventional dendritic cells (cDCs) are found in all tissues and play a key role in immune surveillance. They comprise two major subsets, cDC1 and cDC2, both derived from circulating precursors of cDCs (pre-cDCs), which exited the bone marrow. We show that, in the steady-state mouse, pre-cDCs entering tissues proliferate to give rise to differentiated cDCs, which themselves have residual proliferative capacity. We use multicolor fate mapping of cDC progenitors to show that this results in clones of sister cDCs, most of which comprise a single cDC1 or cDC2 subtype, suggestive of pre-cDC commitment. Upon infection, a surge in the influx of pre-cDCs into the affected tissue dilutes clones and increases cDC numbers. Our results indicate that tissue cDCs can be organized in a patchwork of closely positioned sister cells of the same subset whose coexistence is perturbed by local infection, when the bone marrow provides additional pre-cDCs to meet increased tissue demand

Cholesterol sensor ORP1L contacts the ER protein VAP to control Rab7–RILP–p150Glued and late endosome positioning

Late endosomes (LEs) have characteristic intracellular distributions determined by their interactions with various motor proteins. Motor proteins associated to the dynactin subunit p150Glued bind to LEs via the Rab7 effector Rab7-interacting lysosomal protein (RILP) in association with the oxysterol-binding protein ORP1L. We found that cholesterol levels in LEs are sensed by ORP1L and are lower in peripheral vesicles. Under low cholesterol conditions, ORP1L conformation induces the formation of endoplasmic reticulum (ER)–LE membrane contact sites. At these sites, the ER protein VAP (VAMP [vesicle-associated membrane protein]-associated ER protein) can interact in trans with the Rab7–RILP complex to remove p150Glued and associated motors. LEs then move to the microtubule plus end. Under high cholesterol conditions, as in Niemann-Pick type C disease, this process is prevented, and LEs accumulate at the microtubule minus end as the result of dynein motor activity. These data explain how the ER and cholesterol control the association of LEs with motor proteins and their positioning in cells

Arp2/3 complex interactions and actin network turnover in lamellipodia

Cell migration is initiated by lamellipodia—membrane-enclosed sheets of cytoplasm containing densely packed actin filament networks. Although the molecular details of network turnover remain obscure, recent work points towards key roles in filament nucleation for Arp2/3 complex and its activator WAVE complex. Here, we combine fluorescence recovery after photobleaching (FRAP) of different lamellipodial components with a new method of data analysis to shed light on the dynamics of actin assembly/disassembly. We show that Arp2/3 complex is incorporated into the network exclusively at the lamellipodium tip, like actin, at sites coincident with WAVE complex accumulation. Capping protein likewise showed a turnover similar to actin and Arp2/3 complex, but was confined to the tip. In contrast, cortactin—another prominent Arp2/3 complex regulator—and ADF/cofilin—previously implicated in driving both filament nucleation and disassembly—were rapidly exchanged throughout the lamellipodium. These results suggest that Arp2/3- and WAVE complex-driven actin filament nucleation at the lamellipodium tip is uncoupled from the activities of both cortactin and cofilin. Network turnover is additionally regulated by the spatially segregated activities of capping protein at the tip and cofilin throughout the mesh

Correction:How the COVID-19 pandemic highlights the necessity of animal research (vol 30, pg R1014, 2020)

(Current Biology 30, R1014–R1018; September 21, 2020) As a result of an author oversight in the originally published version of this article, a number of errors were introduced in the author list and affiliations. First, the middle initials were omitted from the names of several authors. Second, the surname of Dr. van Dam was mistakenly written as “Dam.” Third, the first name of author Bernhard Englitz was misspelled as “Bernard” and the surname of author B.J.A. Pollux was misspelled as “Pullox.” Finally, Dr. Keijer's first name was abbreviated rather than written in full. These errors, as well as various errors in the author affiliations, have now been corrected online

Systems microscopy approaches to understand cancer cell migration and metastasis

Cell migration is essential in a number of processes, including wound healing, angiogenesis and cancer metastasis. Especially, invasion of cancer cells in the surrounding tissue is a crucial step that requires increased cell motility. Cell migration is a well-orchestrated process that involves the continuous formation and disassembly of matrix adhesions. Those structural anchor points interact with the extra-cellular matrix and also participate in adhesion-dependent signalling. Although these processes are essential for cancer metastasis, little is known about the molecular mechanisms that regulate adhesion dynamics during tumour cell migration. In this review, we provide an overview of recent advanced imaging strategies together with quantitative image analysis that can be implemented to understand the dynamics of matrix adhesions and its molecular components in relation to tumour cell migration. This dynamic cell imaging together with multiparametric image analysis will help in understanding the molecular mechanisms that define cancer cell migration

Measuring aeolian sand transport using acoustic sensors

Acoustic sensors are frequently used to measure aeolian saltation. Different approaches are used to process the signals from these instruments. The goal of this paper is to describe and discuss a method to measure aeolian saltation with acoustic sensors. In a laboratory experiment, we measured the output from an advanced signal processing scheme on the circuit board of the saltiphone. We use a software implementation of this processing scheme to re-analyse data from four miniphones obtained during a field experiment. It is shown that a set of filters remove background noise outside the frequency spectrum of aeolian saltation (at 8. kHz), whereas signals within this frequency spectrum are amplified. The resulting analogue signal is a proxy of the energy. Using an AC pulse convertor, this signal can be converted into a digital and analogue count signal or an analogue energy signal, using a rectifier and integrator. Spatio-temporal correlation between field deployed miniphones increases by using longer integration times for signal processing. To quantify aeolian grain impact, it is suggested to use the analogue energy output, as this mode is able to detect changes in frequency and amplitude. The analogue and digital count signals are able to detect an increase in frequency, but are not able to detect an increase in signal amplitude. We propose a two-stage calibration scheme consisting of (1) a factory calibration, to set the frequency spectrum of the sensor and (2) a standardized drop-test conducted before and after the experiment to evaluate the response of the sensor

Measuring aeolian sand transport using acoustic sensors

Acoustic sensors are frequently used to measure aeolian saltation. Different approaches are used to process the signals from these instruments. The goal of this paper is to describe and discuss a method to measure aeolian saltation with acoustic sensors. In a laboratory experiment, we measured the output from an advanced signal processing scheme on the circuit board of the saltiphone. We use a software implementation of this processing scheme to re-analyse data from four miniphones obtained during a field experiment. It is shown that a set of filters remove background noise outside the frequency spectrum of aeolian saltation (at 8. kHz), whereas signals within this frequency spectrum are amplified. The resulting analogue signal is a proxy of the energy. Using an AC pulse convertor, this signal can be converted into a digital and analogue count signal or an analogue energy signal, using a rectifier and integrator. Spatio-temporal correlation between field deployed miniphones increases by using longer integration times for signal processing. To quantify aeolian grain impact, it is suggested to use the analogue energy output, as this mode is able to detect changes in frequency and amplitude. The analogue and digital count signals are able to detect an increase in frequency, but are not able to detect an increase in signal amplitude. We propose a two-stage calibration scheme consisting of (1) a factory calibration, to set the frequency spectrum of the sensor and (2) a standardized drop-test conducted before and after the experiment to evaluate the response of the sensor.</p