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Ariel Molly Wilson, Zhuo Shao, Vanessa Grenier, Gaëlle Mawambo, Jean-François Daudelin, Agnieszka Dejda, Frédérique Pilon, Natalija Popovic, Salix Boulet, Célia Parinot, Malika Oubaha, Nathalie Labrecque, Vincent de Guire, Mathieu Laplante, Guillaume Lettre, Florian Sennlaub, Jean-Sebastien Joyal, Michel Meunier, Przemyslaw Sapieha
Obesity gives rise to metabolic complications by mechanisms that are poorly understood. Although chronic inflammatory signaling in adipose tissue is typically associated with metabolic deficiencies linked to excessive weight gain, we identified a subset of neuropilin-1 (NRP1)-expressing myeloid cells that accumulate in adipose tissue and protect against obesity and metabolic syndrome. Ablation of NRP1 in macrophages compromised lipid uptake in these cells, which reduced substrates for fatty acid β-oxidation and shifted energy metabolism of these macrophages toward a more inflammatory glycolytic metabolism...
March 16, 2018: Science Immunology
Tom Zortel, Annette Schmitt-Graeff, Susanne Kirschnek, Georg Häcker
Background: Chlamydial infection frequently causes damage to the female genital tract. The precise mechanisms of chlamydial clearance and tissue damage are unknown but studies suggest immunopathology with a particular role of neutrophils. The goal of this study was to understand the contribution of the immune system, in particular neutrophils. Methods: Using Chlamydia muridarum, we infected mice with a prolonged immune response due to expression of Bcl-2 in haematopoietic cells (Bcl-2-mice), and mice where mature neutrophils are lacking due to the deletion of Mcl-1 in myeloid cells (LysM-cre-mcl-1-flox-mice; Mcl-1-mice)...
March 7, 2018: Journal of Infectious Diseases
Sharmila Nair, Jeremy P Huynh, Vicky Lampropoulou, Ekaterina Loginicheva, Ekaterina Esaulova, Anshu P Gounder, Adrianus C M Boon, Elizabeth A Schwarzkopf, Tara R Bradstreet, Brian T Edelson, Maxim N Artyomov, Christina L Stallings, Michael S Diamond
Immune-Responsive Gene 1 (Irg1) is a mitochondrial enzyme that produces itaconate under inflammatory conditions, principally in cells of myeloid lineage. Cell culture studies suggest that itaconate regulates inflammation through its inhibitory effects on cytokine and reactive oxygen species production. To evaluate the functions of Irg1 in vivo, we challenged wild-type (WT) and Irg1 -/- mice with Mycobacterium tuberculosis ( Mtb ) and monitored disease progression. Irg1 -/- , but not WT, mice succumbed rapidly to Mtb , and mortality was associated with increased infection, inflammation, and pathology...
March 6, 2018: Journal of Experimental Medicine
Christine C Yokoyama, Megan T Baldridge, Daisy W Leung, Guoyan Zhao, Chandni Desai, Ta-Chiang Liu, Vladimir E Diaz-Ochoa, Jeremy P Huynh, Jacqueline M Kimmey, Erica L Sennott, Camaron R Hole, Rachel A Idol, Sunmin Park, Kelly M Storek, Caihong Wang, Seungmin Hwang, Ashley Viehmann Milam, Eric Chen, Tobias Kerrinnes, Michael N Starnbach, Scott A Handley, Indira U Mysorekar, Paul M Allen, Denise M Monack, Mary C Dinauer, Tamara L Doering, Renee M Tsolis, Jonathan E Dworkin, Christina L Stallings, Gaya K Amarasinghe, Craig A Micchelli, Herbert W Virgin
Germline-encoded receptors recognizing common pathogen-associated molecular patterns are a central element of the innate immune system and play an important role in shaping the host response to infection. Many of the innate immune molecules central to these signaling pathways are evolutionarily conserved. LysMD3 is a novel molecule containing a putative peptidoglycan-binding domain that has orthologs in humans, mice, zebrafish, flies, and worms. We found that the lysin motif (LysM) of LysMD3 forms a folded structure and is likely related to a previously described peptidoglycan-binding LysM found in bacteria...
March 1, 2018: Journal of Biological Chemistry
Karl T Debiec, Matthew J Whitley, Leonardus M I Koharudin, Lillian T Chong, Angela M Gronenborn
Multidomain proteins with two or more independently folded functional domains are prevalent in nature. Whereas most multidomain proteins are linked linearly in sequence, roughly one-tenth possess domain insertions where a guest domain is implanted into a loop of a host domain, such that the two domains are connected by a pair of interdomain linkers. Here, we characterized the influence of the interdomain linkers on the structure and dynamics of a domain-insertion protein in which the guest LysM domain is inserted into a central loop of the host CVNH domain...
February 27, 2018: Biophysical Journal
Soichi Sano, Kosei Oshima, Ying Wang, Susan MacLauchlan, Yasufumi Katanasaka, Miho Sano, María A Zuriaga, Minoru Yoshiyama, David Goukassian, Matthew A Cooper, José J Fuster, Kenneth Walsh
BACKGROUND: Recent studies have shown that hematopoietic stem cells can undergo clonal expansion secondary to somatic mutations in leukemia-related genes, thus leading to an age-dependent accumulation of mutant leukocytes in the blood. This somatic mutation-related clonal hematopoiesis is common in healthy older individuals, but it has been associated with an increased incidence of future cardiovascular disease. The epigenetic regulator TET2 is frequently mutated in blood cells of individuals exhibiting clonal hematopoiesis...
February 27, 2018: Journal of the American College of Cardiology
Mei Chen, Jiawu Zhao, Imran H A Ali, Stephen Marry, Josy Augustine, Mohajeet Bhuckory, Aisling Lynch, Adrien Kissenpfennig, Heping Xu
The suppressor of cytokine signaling protein 3 (SOCS3) critically controls immune cell activation, although its role in macrophage polarization and function remains controversial. Using experimental autoimmune uveoretinitis (EAU) as a model, we show that inflammation-mediated retinal degeneration is exaggerated and retinal angiogenesis is accelerated in mice with SOCS3 deficiency in myeloid cells (LysMCre/+ SOCS3fl/fl ). At the acute stage of EAU, the population of infiltrating neutrophils was increased and the population of macrophages decreased in LysMCre/+ SOCS3fl/fl mice compared with that in wild-type (WT) mice...
February 13, 2018: American Journal of Pathology
Tanja Schönfelder, Moritz Brandt, Sabine Kossmann, Tanja Knopp, Thomas Münzel, Ulrich Walter, Susanne H Karbach, Philip Wenzel
The role of leukocytes in deep vein thrombosis (DVT) resolution is incompletely understood. We determined how depletion of lysozyme positive (LysM + ) cells and a switched-off type 1 immune response influences thrombus resolution. DVT was induced in 12-week-old male mice by inferior vena cava (IVC) stenosis. Toxin mediated depletion of myeloid cells improved thrombus resolution in mice with Cre-inducible expression of the diphtheria toxin receptor in LysM + cells. This correlated with decreased CD45 + cells, a population shift of Gr-1 + to Gr-1 - CD11b + myelomonocytic cells (flow cytometry) and an increase in CC-chemokine ligand 2, interleukin-4 and interleukin-10 mRNA expressions...
February 14, 2018: Scientific Reports
Sang A Park, Young Ho Choe, Sung Hwan Lee, Young-Min Hyun
Sepsis is a type of severe infection that can cause organ failure and tissue damage. Although the mortality and morbidity rates associated with sepsis are extremely high, no direct treatment or organ-related mechanism has been examined in detail in real time. The liver is the key organ that manages toxins and infections in the human body. Herein, we aimed to perform intravital imaging of mouse liver after induction of endotoxemia in order to track the motility of immune cells, such as neutrophils and liver capsular macrophages (LCMs)...
February 6, 2018: Journal of Visualized Experiments: JoVE
Hamida Beghalem, Khadidja Aliliche, Ahmed Landoulsi
In legumes, LysM domains of receptors-like kinases (RLKs) mediate rhizobial NFs perception; which are required for infection and establishment of symbiosis without triggering the host immune response. In this study, we identify the LysM extracellular domain sequences of putative Sulla species Nod factor receptors (S. pallida, S. capitata and S. coronaria). The Blast search displayed high identity percentages with genes encoding LjNFR5-like of several legumes. Phylogenetic trees were built using the partial nod factor receptor and predicted amino acid sequences, which grouped Sulla in a separate clade...
February 2, 2018: Microbial Pathogenesis
Patricia Romero, Sergio G Bartual, Mathias Schmelcher, Chaim Glück, Juan A Hermoso, Martin J Loessner
Endolysins are bacteriophage-encoded peptidoglycan hydrolases that specifically degrade the bacterial cell wall at the end of the phage lytic cycle. They feature a distinct modular architecture, consisting of enzymatically active domains (EADs) and cell wall binding domains (CBDs). Structural analysis of the complete enzymes or individual domains is required for better understanding the mechanisms of peptidoglycan degradation and provides guidelines for the rational design of chimeric enzymes. We here report the crystal structure of the EAD of PlyP40, a member of the GH-25 family of glycosyl hydrolases, and the first muramidase reported for Listeria phages...
February 5, 2018: Molecular Microbiology
Katsuhiko Shinomiya, Mayumi Ueta, Shigeru Kinoshita
Chronic dry eye is an increasingly prevalent condition worldwide, with resulting loss of visual function and quality of life. Relevant, repeatable, and stable animal models of dry eye are still needed. We have developed an improved surgical mouse model for dry eye based on severe aqueous fluid deficiency, by excising both the exorbital and intraorbital lacrimal glands (ELG and ILG, respectively) of mice. After ELG plus ILG excision, dry eye symptoms were evaluated using fluorescein infiltration observation, tear production measurement, and histological evaluation of ocular surface...
January 24, 2018: Scientific Reports
Jeremy Lagrange, Sabine Kossmann, Klytaimnistra Kiouptsi, Philip Wenzel
Epifluorescence intravital video microscopy (IVM) of blood vessels is an established method to evaluate the activation of immune cells and their ability to role and adhere to the endothelial layer. Visualization of circulating cells by injection of fluorescent dyes or fluorophore-coupled antibodies is commonly used. Alternatively, fluorescent reporter mice can be used. Interactions of leukocytes, in particular lysozyme M+ (LysM+) monocytes, with the vessel wall play pivotal roles in promoting vascular dysfunction and arterial hypertension...
January 4, 2018: Journal of Visualized Experiments: JoVE
Hongmei Li, Sang Jun Han, Mihwa Kim, Ahyeon Cho, Yewoon Choi, Vivette Denise D'Agati, H Thomas Lee
We previously demonstrated that kidney peptidylarginine deiminase-4 (PAD4) plays a critical role in ischemic acute kidney injury (AKI) in mice by promoting renal tubular inflammation and neutrophil infiltration. Although the role of PAD4 in granulocytes including neutrophils is well known, we surprisingly observed profound renal proximal tubular PAD4 induction after renal ischemia and reperfusion (IR) injury. Here we tested the hypothesis that renal proximal tubular PAD4 rather than myeloid cell lineage PAD4 plays a critical role in exacerbating ischemic AKI by utilizing mice lacking PAD4 in renal proximal tubules (PAD4ff PEPCK Cre mice) or in granulocytes (PAD4ff LysM Cre mice)...
January 3, 2018: American Journal of Physiology. Renal Physiology
Yu Gao, Juan Ignacio Basile, Cajsa Classon, Dolores Gavier-Widen, Akihiko Yoshimura, Berit Carow, Martin E Rottenberg
STAT3 is a master regulator of the immune responses. Here we show that M. tuberculosis-infected stat3fl/fl lysm cre mice, defective for STAT3 in myeloid cells, contained lower bacterial load in lungs and spleens, reduced granuloma extension but higher levels of pulmonary neutrophils. STAT3-deficient macrophages showed no improved control of intracellular mycobacterial growth. Instead, protection associated to elevated ability of stat3fl/fl lysm cre antigen-presenting cells (APCs) to release IL-6 and IL-23 and to stimulate IL-17 secretion by mycobacteria-specific T cells...
January 16, 2018: PLoS Pathogens
Rongguang Zhang, Chen Wang, Wenbin Cheng, Guangcai Duan, Qingfeng Shi, Shuaiyin Chen, Qingtang Fan
OBJECTIVE: To develop a safe and effective oral vaccine against Helicobacter pylori using its HpaA protein expressed in Lactococcus lactis. RESULTS: The gene encoding HpaA was obtained by PCR and ligated to pNZ8110-lysM following digestion with NaeI + SphI. The recombinant plasmid was transferred into E. coli for multiplication, and then into L. lactis. The recombinant L. lactis was induced to express HpaA, resulting in two products of 29 and 25 kDa, both of which yielded positive immunoreaction with mouse antisera against H...
January 3, 2018: Biotechnology Letters
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
Hiromasa Wakui, Sayaka Fuseya, Riku Suzuki, Miki Shimbo, Risa Okada, Mitchito Hamada, Akihiro Kuno, Kozue Hagiwara, Takashi Sato, Hisashi Narimatsu, Takashi Kudo, Satoru Takahashi
The core 1 β1,3-galactosyltransferase-specific molecular chaperon (Cosmc) is essential for the synthesis of the core 1 structure of mucin-type O-glycans. To clarify the physiological role of core 1-derived O-glycans in macrophages, we exploited the LysM-Cre transgene to generate a conditional Cosmc mutant allele (conditional Cosmc knockout; cKO) in myeloid cells. cKO mice developed normally with no gross phenotypic abnormalities or abnormal peripheral blood counts. Resident peritoneal macrophages (rpMacs) of cKO mice exhibited impaired engulfment of apoptotic cells but showed normal macrophage differentiation and counts...
December 13, 2017: Biochemical and Biophysical Research Communications
Alexandra L McCubbrey, Kristen C Allison, Alisa B Lee-Sherick, Claudia V Jakubzick, William J Janssen
Conditional and inducible Cre-loxP systems are used to target gene deletion to specific cell lineages and tissues through promoter-restricted expression of the bacterial DNA recombinase, Cre. Although Cre-loxP systems are widely used to target gene deletion in lung macrophages, limited data are published on the specificity and efficiency of "macrophage targeting" Cre lines. Using R26-stopfl/fl -TdTomato and tetOn-GFP reporter lines, we assessed the specificity and efficiency of four commercially available Cre driver lines that are often considered "macrophage specific...
2017: Frontiers in Immunology
Koji Shido, Deebly Chavez, Zhongwei Cao, Jane Ko, Shahin Rafii, Bi-Sen Ding
In mammals, the livers regenerate after chemical injury or resection of hepatic lobe by hepatectomy. How liver regeneration is initiated after mass loss remains to be defined. Here, we report that following liver injury, activated platelets deploy SDF-1 and VEGF-A to stimulate CXCR7+ liver sinusoidal endothelial cell (LSEC) and VEGFR1+ myeloid cell, orchestrating hepatic regeneration. After carbon tetrachloride (CCl4 ) injection or hepatectomy, platelets and CD11b+ VEGFR1+ myeloid cells were recruited LSEC, and liver regeneration in both models was impaired in thrombopoietin-deficient ( Thpo -/- ) mice lacking circulating platelets...
2017: Signal Transduction and Targeted Therapy
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