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Intravital microscopy

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https://www.readbyqxmd.com/read/29342141/monitoring-t-cell-dendritic-cell-interactions-in-vivo-by-intercellular-enzymatic-labelling
#1
Giulia Pasqual, Aleksey Chudnovskiy, Jeroen M J Tas, Marianna Agudelo, Lawrence D Schweitzer, Ang Cui, Nir Hacohen, Gabriel D Victora
Interactions between different cell types are essential for multiple biological processes, including immunity, embryonic development and neuronal signalling. Although the dynamics of cell-cell interactions can be monitored in vivo by intravital microscopy, this approach does not provide any information on the receptors and ligands involved or enable the isolation of interacting cells for downstream analysis. Here we describe a complementary approach that uses bacterial sortase A-mediated cell labelling across synapses of immune cells to identify receptor-ligand interactions between cells in living mice, by generating a signal that can subsequently be detected ex vivo by flow cytometry...
January 17, 2018: Nature
https://www.readbyqxmd.com/read/29335647/aged-polymorphonuclear-leukocytes-cause-fibrotic-interstitial-lung-disease-in-the-absence-of-regulation-by-b-cells
#2
Jung Hwan Kim, John Podstawka, Yuefei Lou, Lu Li, Esther K S Lee, Maziar Divangahi, Björn Petri, Frank R Jirik, Margaret M Kelly, Bryan G Yipp
Pulmonary immunity requires tight regulation, as interstitial inflammation can compromise gas exchange and lead to respiratory failure. Here we found a greater number of aged CD11bhiL-selectinloCXCR4+ polymorphonuclear leukocytes (PMNs) in lung vasculature than in the peripheral circulation. Using pulmonary intravital microscopy, we observed lung PMNs physically interacting with B cells via β2 integrins; this initiated neutrophil apoptosis, which led to macrophage-mediated clearance. Genetic deletion of B cells led to the accumulation of aged PMNs in the lungs without systemic inflammation, which caused pathological fibrotic interstitial lung disease that was attenuated by the adoptive transfer of B cells or depletion of PMNs...
January 15, 2018: Nature Immunology
https://www.readbyqxmd.com/read/29324806/immunocompetent-host-develops-mild-intestinal-inflammation-in-acute-infection-with-toxoplasma-gondii
#3
Paulo da Silva Watanabe, Aline Rosa Trevizan, Saulo Euclides Silva-Filho, Marcelo Biondaro Góis, João Luiz Garcia, Roberto Kenji Nakamura Cuman, Ana Cristina Breithaupt-Faloppa, Débora de Mello Gonçales Sant Ana, Gessilda de Alcantara Nogueira de Melo
Toxoplasma gondii (T. gondii) is the causative agent of toxoplasmosis, common zoonosis among vertebrates and high incidence worldwide. During the infection, the parasite needs to transpose the intestinal barrier to spread throughout the body, which may be a trigger for an inflammatory reaction. This work evaluated the inflammatory alterations of early T. gondii infection in peripheral blood cells, in the mesenteric microcirculation, and small intestinal tissue by measurement of MPO (myeloperoxidase) activity and NO (nitric oxide) level in rats...
2018: PloS One
https://www.readbyqxmd.com/read/29320154/high-precision-photothermal-ablation-using-biocompatible-palladium-nanoparticles-and-laser-scanning-microscopy
#4
Belén Rubio Ruiz, Ana Perez-Lopez, Thomas Bray, Martin Lee, Alan Serrels, Martin Prieto, Manuel Arruebo, Neil O Carragher, Victor Sebastian, Asier Unciti-Broceta
Herein we report a straightforward method for the scalable preparation of Pd nanoparticles (Pd-NPs) with reduced inherent cytotoxicity and high photothermal conversion capacity. These Pd-NPs are rapidly taken up by cells and able to kill labelled cancer cells upon short exposure to near infrared (NIR) light. Following cell treatment with Pd-NPs, ablated areas were patterned with high precision by laser scanning microscopy, allowing to perform cell migration assays with unprecedented accuracy. Using coherent Raman microscopy, cells containing Pd-NPs were simultaneously ablated and imaged...
January 10, 2018: ACS Applied Materials & Interfaces
https://www.readbyqxmd.com/read/29311694/intravital-mucosal-imaging-of-cd8-resident-memory-t-cells-shows-tissue-autonomous-recall-responses-that-amplify-secondary-memory
#5
Lalit K Beura, Jason S Mitchell, Emily A Thompson, Jason M Schenkel, Javed Mohammed, Sathi Wijeyesinghe, Raissa Fonseca, Brandon J Burbach, Heather D Hickman, Vaiva Vezys, Brian T Fife, David Masopust
CD8+ T cell immunosurveillance dynamics influence the outcome of intracellular infections and cancer. Here we used two-photon intravital microscopy to visualize the responses of CD8+ resident memory T cells (TRM cells) within the reproductive tracts of live female mice. We found that mucosal TRM cells were highly motile, but paused and underwent in situ division after local antigen challenge. TRM cell reactivation triggered the recruitment of recirculating memory T cells that underwent antigen-independent TRM cell differentiation in situ...
January 8, 2018: Nature Immunology
https://www.readbyqxmd.com/read/29311541/yolk-sac-macrophage-progenitors-traffic-to-the-embryo-during-defined-stages-of-development
#6
C Stremmel, R Schuchert, F Wagner, R Thaler, T Weinberger, R Pick, E Mass, H C Ishikawa-Ankerhold, A Margraf, S Hutter, R Vagnozzi, S Klapproth, J Frampton, S Yona, C Scheiermann, J D Molkentin, U Jeschke, M Moser, M Sperandio, S Massberg, F Geissmann, C Schulz
Tissue macrophages in many adult organs originate from yolk sac (YS) progenitors, which invade the developing embryo and persist by means of local self-renewal. However, the route and characteristics of YS macrophage trafficking during embryogenesis are incompletely understood. Here we show the early migration dynamics of YS-derived macrophage progenitors in vivo using fate mapping and intravital microscopy. From embryonic day 8.5 (E8.5) CX3CR1+ pre-macrophages are present in the mouse YS where they rapidly proliferate and gain access to the bloodstream to migrate towards the embryo...
January 8, 2018: Nature Communications
https://www.readbyqxmd.com/read/29300390/a-surgical-orthotopic-organoid-transplantation-approach-in-mice-to-visualize-and-study-colorectal-cancer-progression
#7
Arianna Fumagalli, Saskia J E Suijkerbuijk, Harry Begthel, Evelyne Beerling, Koen C Oost, Hugo J Snippert, Jacco van Rheenen, Jarno Drost
Most currently available colorectal cancer (CRC) mouse models are not suitable for studying progression toward the metastatic stage. Recently, establishment of tumor organoid lines, either from murine CRC models or patients, and the possibility of engineering them with genome-editing technologies, have provided a large collection of tumor material faithfully recapitulating phenotypic and genetic heterogeneity of native tumors. To study tumor progression in the natural in vivo environment, we developed an orthotopic approach based on transplantation of CRC organoids into the cecal epithelium...
February 2018: Nature Protocols
https://www.readbyqxmd.com/read/29298810/highly-angiogenic-non-thrombogenic-bone-marrow-mononuclear-cells-derived-spheroids-in-intraportal-islet-transplantation
#8
Bae Jun Oh, Sang-Man Jin, Yoonha Hwang, Jin Myung Choi, Han-Sin Lee, Gyuri Kim, Geunsoo Kim, Hyo Jun Park, Pilhan Kim, Sung Joo Kim, Jae Hyeon Kim
Highly angiogenic bone marrow mononuclear cells-derived spheroids (BM-spheroids), formed by selective proliferation of the CD31+CD14+CD34+ monocyte subset via 3D culture, had robust angiogenetic capacity in rodent syngeneic renal subcapsular islet transplantation. We wondered whether the efficacy of BM-spheroids could be demonstrated in clinically-relevant intraportal islet transplantation models without increasing the risk of portal thrombosis. The thrombogenic potential of intraportally infused BM-spheroids was compared with that of mesenchymal stromal cells (MSCs) and MSC-spheroids...
January 3, 2018: Diabetes
https://www.readbyqxmd.com/read/29287721/intravital-imaging-of-neutrophil-recruitment-in-intestinal-ischemia-reperfusion-injury
#9
Shintaro Hashimoto, Masaki Honda, Takayuki Takeichi, Masataka Sakisaka, Yasuko Narita, Daiki Yoshii, Keiichi Uto, Seisuke Sakamoto, Yukihiro Inomata
BACKGROUND: Neutrophils are known to be key players in innate immunity. Activated neutrophils induce local inflammation, which results in pathophysiologic changes during intestinal ischemia-reperfusion injury (IRI). However, most studies have been based on static assessments, and few have examined real-time intravital neutrophil recruitment. We herein report a method for imaging and evaluating dynamic changes in the neutrophil recruitment in intestinal IRI using two-photon laser scanning microscopy (TPLSM)...
January 15, 2018: Biochemical and Biophysical Research Communications
https://www.readbyqxmd.com/read/29287222/in-vivo-imaging-of-adaptive-immune-responses-to-viruses
#10
REVIEW
Marco De Giovanni, Matteo Iannacone
Viral infections represent a major threat for mankind. The adaptive immune system plays a key role in both viral clearance and disease pathogenesis, and, accordingly, understanding how lymphocytes interact with different viruses is critical to design more effective vaccination and therapeutic strategies. The recent advent of intravital microscopy has enabled the real-time visualization of the complex interplay between viruses and the ensuing adaptive immune response in living organisms. Here, we will review the most significant recent insights on antiviral adaptive immune responses obtained through intravital imaging...
December 26, 2017: Current Opinion in Virology
https://www.readbyqxmd.com/read/29286871/in-vivo-modeling-of-docosahexaenoic-acid-and-eicosapentaenoic-acid-mediated-inhibition-of-both-platelet-function-and-accumulation-in-arterial-thrombi
#11
Reheman Adili, Ellen M Voigt, Jordan L Bormann, Kaitlynn N Foss, Luke J Hurley, Evan S Meyer, Amber J Veldman, Katherine A Mast, Joshua L West, Sidney W Whiteheart, Michael Holinstat, Mark K Larson
Omega-3 polyunsaturated fatty acids (n-3 PUFAs) are associated with a variety of cellular alterations that mitigate cardiovascular disease. However, pinpointing the positive therapeutic effects is challenging due to inconsistent clinical trial results and overly simplistic in vitro studies. Here we aimed to develop realistic models of n-3 PUFA effects on platelet function so that preclinical results can better align with and predict clinical outcomes. Human platelets incubated with the n-3 PUFAs docosahexaenoic acid and eicosapentaenoic acid were stimulated with agonist combinations mirroring distinct regions of a growing thrombus...
December 29, 2017: Platelets
https://www.readbyqxmd.com/read/29286428/stenosis-of-the-inferior-vena-cava-a-murine-model-of-deep-vein-thrombosis
#12
Holly Payne, Alexander Brill
Deep vein thrombosis (DVT) and its devastating complication, pulmonary embolism, are a severe health problem with high mortality. Mechanisms of thrombus formation in veins remain obscure. Lack of mobility (e.g., after surgery or long-haul flights) is one of the main factors leading to DVT. The pathophysiological consequence of the lack of mobility is blood flow stagnation in venous valves. Here, a model is described that mimics such flow disturbance as a thrombosis-driving factor. In this model, partial flow restriction (stenosis) in the inferior vena cava (IVC) is created...
December 22, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/29285816/arterialized-collateral-capillaries-progress-from-non-reactive-to-capable-of-increasing-perfusion-in-an-ischemic-tree
#13
Kenneth H Gouin, Sara K Hellstrom, Lindsay E Clegg, Joshua Cutts, Feilim Mac Gabhann, Trevor R Cardinal
OBJECTIVE: Collateral capillary arteriogenenesis (CCA), outward remodeling of capillaries that anastomose two arteriolar trees with different parent feed arteries, may represent a therapeutic target for patients who lack collaterals. Arterialized collateral capillaries (ACCs) can reperfuse an ischemic tree, but their functional capacity is unknown. Therefore, we determined if ACCs mature into resistance vessels that regulate blood flow following arterial occlusion. METHODS: We ligated the lateral spinotrapezius feed artery in Balb/C mice, which induces CCA...
December 28, 2017: Microcirculation: the Official Journal of the Microcirculatory Society, Inc
https://www.readbyqxmd.com/read/29281816/tumor-associated-macrophages-derived-from-circulating-inflammatory-monocytes-degrade-collagen-through-cellular-uptake
#14
Daniel Hargbøl Madsen, Henrik Jessen Jürgensen, Majken Storm Siersbæk, Dorota Ewa Kuczek, Loretta Grey Cloud, Shihui Liu, Niels Behrendt, Lars Grøntved, Roberto Weigert, Thomas Henrik Bugge
Physiologic turnover of interstitial collagen is mediated by a sequential pathway in which collagen is fragmented by pericellular collagenases, endocytosed by collagen receptors, and routed to lysosomes for degradation by cathepsins. Here, we use intravital microscopy to investigate if malignant tumors, which are characterized by high rates of extracellular matrix turnover, utilize a similar collagen degradation pathway. Tumors of epithelial, mesenchymal, or neural crest origin all display vigorous endocytic collagen degradation...
December 26, 2017: Cell Reports
https://www.readbyqxmd.com/read/29276143/inhibition-of-endosteal-vascular-niche-remodeling-rescues-hematopoietic-stem-cell-loss-in-aml
#15
Delfim Duarte, Edwin D Hawkins, Olufolake Akinduro, Heather Ang, Katia De Filippo, Isabella Y Kong, Myriam Haltalli, Nicola Ruivo, Lenny Straszkowski, Stephin J Vervoort, Catriona McLean, Tom S Weber, Reema Khorshed, Chiara Pirillo, Andrew Wei, Saravana K Ramasamy, Anjali P Kusumbe, Ken Duffy, Ralf H Adams, Louise E Purton, Leo M Carlin, Cristina Lo Celso
Bone marrow vascular niches sustain hematopoietic stem cells (HSCs) and are drastically remodeled in leukemia to support pathological functions. Acute myeloid leukemia (AML) cells produce angiogenic factors, which likely contribute to this remodeling, but anti-angiogenic therapies do not improve AML patient outcomes. Using intravital microscopy, we found that AML progression leads to differential remodeling of vasculature in central and endosteal bone marrow regions. Endosteal AML cells produce pro-inflammatory and anti-angiogenic cytokines and gradually degrade endosteal endothelium, stromal cells, and osteoblastic cells, whereas central marrow remains vascularized and splenic vascular niches expand...
December 6, 2017: Cell Stem Cell
https://www.readbyqxmd.com/read/29250746/neutrophils-in-the-lung-the-first-responders
#16
REVIEW
Gurpreet Kaur Aulakh
Neutrophils, constituting the first line of defense, perform vital functions during immune surveillance. A key feature that assists in their prompt response to an inflammatory signal is rapid migration to the affected site. They are normally short-lived but can be activated to live longer under the influence of an inflammatory stimulus. They can, thereby, release their toxic granule contents that are differentially housed inside the cytoplasm. Although these events are well characterized in the peripheral circulation, we are still far from fully understanding their recruitment in lungs...
December 18, 2017: Cell and Tissue Research
https://www.readbyqxmd.com/read/29249807/viral-subversion-of-b-cell-responses-within-secondary-lymphoid-organs
#17
REVIEW
Mirela Kuka, Matteo Iannacone
Antibodies play a crucial role in virus control. The production of antibodies requires virus-specific B cells to encounter viral antigens in lymph nodes, become activated, interact with different immune cells, proliferate and enter specific differentiation programmes. Each step occurs in distinct lymph node niches, requiring a coordinated migration of B cells between different subcompartments. The development of multiphoton intravital microscopy has enabled researchers to begin to elucidate the precise cellular and molecular events by which lymph nodes coordinate humoral responses...
December 18, 2017: Nature Reviews. Immunology
https://www.readbyqxmd.com/read/29241626/c5a-receptor-1-promotes-autoimmunity-neutrophil-dysfunction-and-injury-in-experimental-anti-myeloperoxidase-glomerulonephritis
#18
Jonathan Dick, Poh-Yi Gan, Sharon L Ford, Dragana Odobasic, Maliha A Alikhan, Sven H Loosen, Pam Hall, Clare L Westhorpe, Anqi Li, Joshua D Ooi, Trent M Woodruff, Charles R Mackay, A Richard Kitching, Michael J Hickey, Stephen R Holdsworth
The prospects for complement-targeted therapy in ANCA-associated vasculitis have been enhanced by a recent clinical trial in which C5a receptor 1 (C5aR1) inhibition safely replaced glucocorticoids in induction treatment. C5aR1 primes neutrophils for activation by anti-neutrophil cytoplasmic antibody (ANCA) and is therefore required in models of glomerulonephritis induced by anti-myeloperoxidase antibody. Although humoral and cellular autoimmunity play essential roles in ANCA-associated vasculitis, a role for C5aR1 in these responses has not been described...
December 11, 2017: Kidney International
https://www.readbyqxmd.com/read/29238016/-imaging-of-bone-and-joint-destruction
#19
Junichi Kikuta, Masaru Ishii
  Osteoclasts are bone-resorbing giant polykaryons that differentiate from mononuclear macrophage/monocyte-lineage hematopoietic precursors. We have originally established an advanced imaging system for visualizing in vivo behavior of osteoclasts and their precursors with intravital two-photon microscopy. By means of the system, we found that sphingosine-1-phosphate, a lipid mediator enriched in blood, controlled the migratory behavior of osteoclast precursors. We also developed pH-sensing chemical fluorescent probes to detect localized acidification by bone-resorbing osteoclasts on the bone surface in vivo, and identified two distinct functional states of differentiated osteoclasts, 'bone-resorptive' and 'non-resorptive'...
2017: Nihon Rinshō Men'eki Gakkai Kaishi, Japanese Journal of Clinical Immunology
https://www.readbyqxmd.com/read/29222804/preparation-and-characterization-of-biocompatible-chitosan-nanoparticles-for-targeted-brain-delivery-of-peptides
#20
Muge Yemisci, Secil Caban, Eduardo Fernandez-Megia, Yilmaz Capan, Patrick Couvreur, Turgay Dalkara
Here, we describe a nanocarrier system that can transfer chitosan nanoparticles loaded with either small peptides such as the caspase inhibitor Z-DEVD-FMK or a large peptide like basic fibroblast growth factor across the blood-brain barrier. The nanoparticles are selectively directed to the brain and are not measurably taken up by the liver and spleen. Intravital fluorescent microscopy provides an opportunity to study the penetration kinetics of nanoparticles loaded with fluorescent agents such as Nile red...
2018: Methods in Molecular Biology
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