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T cell facs analysis

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https://www.readbyqxmd.com/read/28417335/poor-prognosis-of-patients-with-triple-negative-breast-cancer-can-be-stratified-by-rank-and-rankl-dual-expression
#1
Monica E Reyes, Takeo Fujii, Daniel Branstetter, Savitri Krishnamurthy, Hiroko Masuda, Xiaoping Wang, James M Reuben, Wendy A Woodward, Beatrice J Edwards, Gabriel N Hortobagyi, Debu Tripathy, William C Dougall, Bedrich L Eckhardt, Naoto T Ueno
PURPOSE: As clinical studies have correlated RANK expression levels with survival in breast cancer, and that RANK signaling is dependent on its cognate ligand RANKL, we hypothesized that dual protein expression further stratifies the poor outcome in TNBC. METHODS: RANK mRNA and protein expression was evaluated in TNBC using genomic databases, cell lines and in a tissue microarray of curated primary tumor samples derived from 87 patients with TNBC. RANK expression was evaluated either by Mann-Whitney U test on log-normalized gene expression data or by Student's t test on FACS data...
April 17, 2017: Breast Cancer Research and Treatment
https://www.readbyqxmd.com/read/28396518/rapamycin-efficiently-promotes-cardiac-differentiation-of-mouse-embryonic-stem-cells
#2
Qin Lu, Zhuqing Jia, Yang Wang, Weiping Wang, Zhe Yang, Tao Li, Yuyao Tian, Ping Chen, Kangtao Ma, Yinan Liu, Chunyan Zhou
To investigate the effects of the rapamycin on cardiac differentiation, murine ESCs was induced to cardiomyocytes by 10(-4) M ascorbic acid (AA), 20 nM rapamycin alone or 0.1‰ solvent dimethyl sulfoxide (DMSO). We found that rapamycin alone was insufficient to initiate cardiomyogenesis. Then the ESCs was treated with AA and rapamycin (20 nM) or AA and DMSO (0.1‰) as a control. Compared with control, mESCs treated with rapamycin (20 nM) and AA yielded a significantly higher percentage of cardiomyocytes, as confirmed by percentage of beating EBs, the immunofluorescence and FACS analysis...
April 10, 2017: Bioscience Reports
https://www.readbyqxmd.com/read/28356009/ecpirm-a-potential-therapeutic-agent-for-cutaneous-t-cell-lymphoma-inhibits-cell-proliferation-and-promotes-apoptosis-via-a-jak-stat-pathway-the-biological-effect-of-ecpirm-in-ctcl-cell
#3
Hua Yang, Pengcheng Ma, Yuping Cao, Mengli Zhang, Lingjun Li, Jun Wei, Lei Tao, Kun Qian
BACKGROUND: Retinoids are important agents for treatment of cutaneous T-cell lymphomas (CTCL). But side effects and drug resistance caused by activation of RAR/RXR limited their clinical application. So it is urgent to develop new agents fight against CTCL. ECPIRM, a 13-cis retinoic acid derivative, was reported that it inhibited proliferation and induced apoptosis of SCL-1 cells. OBJECTIVE: To evaluated the biological activities and mechanisms of ECPIEM. METHODS: Effect of ECPIRM on cell proliferation was determined by MTT assay and Trypan blue exclusion assay while FACS analysis was used to detect changes of cell cycle and apoptosis in HUT78 cells...
March 27, 2017: Anti-cancer Agents in Medicinal Chemistry
https://www.readbyqxmd.com/read/28327575/analysis-of-exome-data-for-4293-trios-suggests-gpi-anchor-biogenesis-defects-are-a-rare-cause-of-developmental-disorders
#4
Alistair T Pagnamenta, Yoshiko Murakami, John M Taylor, Consuelo Anzilotti, Malcolm F Howard, Venessa Miller, Diana S Johnson, Shereen Tadros, Sahar Mansour, I Karen Temple, Rachel Firth, Elisabeth Rosser, Rachel E Harrison, Bronwen Kerr, Niko Popitsch, Taroh Kinoshita, Jenny C Taylor, Usha Kini
Over 150 different proteins attach to the plasma membrane using glycosylphosphatidylinositol (GPI) anchors. Mutations in 18 genes that encode components of GPI-anchor biogenesis result in a phenotypic spectrum that includes learning disability, epilepsy, microcephaly, congenital malformations and mild dysmorphic features. To determine the incidence of GPI-anchor defects, we analysed the exome data from 4293 parent-child trios recruited to the Deciphering Developmental Disorders (DDD) study. All probands recruited had a neurodevelopmental disorder...
March 22, 2017: European Journal of Human Genetics: EJHG
https://www.readbyqxmd.com/read/28321209/treatment-of-leishmania-leishmania-amazonensis-infected-mice-with-a-combination-of-a-palladacycle-complex-and-heat-killed-propionibacterium-acnes-triggers-protective-cellular-immune-responses
#5
Carolina S Paladi, Danielle A M da Silva, Priscila D Motta, Daniel M Garcia, Daniela Teixeira, Ieda M Longo-Maugéri, Simone Katz, Clara L Barbiéri
Palladacycle complex DPPE 1.2 was previously reported to inhibit the in vitro and in vivo infection by Leishmania (Leishmania) amazonensis. The aim of the present study was to compare the effect of DPPE 1.2, in association with heat-killed Propionibacterium acnes, on L. (L.) amazonensis infection in two mouse strains, BALB/c and C57BL/6, and to evaluate the immune responses of the treated animals. Foot lesions of L. (L.) amazonensis-infected mice were injected with DPPE 1.2 alone, or associated with P. acnes as an adjuvant...
2017: Frontiers in Microbiology
https://www.readbyqxmd.com/read/28236766/oxidative-stress-metabolomics-profiling-and-mechanism-of-local-anesthetic-induced-cell-death-in-yeast
#6
Cory H T Boone, Ryan A Grove, Dana Adamcova, Javier Seravalli, Jiri Adamec
The World Health Organization designates lidocaine as an essential medicine in healthcare, greatly increasing the probability of human exposure. Its use has been associated with ROS generation and neurotoxicity. Physiological and metabolomic alterations, and genetics leading to the clinically observed adverse effects have not been temporally characterized. To study alterations that may lead to these undesirable effects, Saccharomyces cerevisiae grown on aerobic carbon sources to stationary phase was assessed over 6h...
February 3, 2017: Redox Biology
https://www.readbyqxmd.com/read/28202731/microfluidic-droplet-platform-for-ultrahigh-throughput-single-cell-screening-of-biodiversity
#7
Stanislav S Terekhov, Ivan V Smirnov, Anastasiya V Stepanova, Tatyana V Bobik, Yuliana A Mokrushina, Natalia A Ponomarenko, Alexey A Belogurov, Maria P Rubtsova, Olga V Kartseva, Marina O Gomzikova, Alexey A Moskovtsev, Anton S Bukatin, Michael V Dubina, Elena S Kostryukova, Vladislav V Babenko, Maria T Vakhitova, Alexander I Manolov, Maja V Malakhova, Maria A Kornienko, Alexander V Tyakht, Anna A Vanyushkina, Elena N Ilina, Patrick Masson, Alexander G Gabibov, Sidney Altman
Ultrahigh-throughput screening (uHTS) techniques can identify unique functionality from millions of variants. To mimic the natural selection mechanisms that occur by compartmentalization in vivo, we developed a technique based on single-cell encapsulation in droplets of a monodisperse microfluidic double water-in-oil-in-water emulsion (MDE). Biocompatible MDE enables in-droplet cultivation of different living species. The combination of droplet-generating machinery with FACS followed by next-generation sequencing and liquid chromatography-mass spectrometry analysis of the secretomes of encapsulated organisms yielded detailed genotype/phenotype descriptions...
March 7, 2017: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/28138512/aging-affects-bone-marrow-macrophage-polarization-relevance-to-bone-healing
#8
E Gibon, F Loi, Luis A Córdova, J Pajarinen, T Lin, L Lu, A Nabeshima, Z Yao, Stuart B Goodman
Macrophages are an important component of the inflammatory cascade by initiating and modulating the processes leading to tissue regeneration and bone healing. Depending on the local environment, macrophages can be polarized into M1 (pro-inflammatory) or M2 (anti-inflammatory) phenotypes. In order to assess the effects of aging on macrophage function, bone marrow macrophage polarization using primary bone marrow macrophages (BMMs) from young (8 weeks old) and aged (72 weeks old) wild-type male C57BL/6J mice was analyzed...
June 2016: Regenerative Engineering and Translational Medicine
https://www.readbyqxmd.com/read/28109145/-short-term-use-of-rapamycin-combined-with-tregs-prolongs-mouse-cardiac-graft-survival-but-attenuates-recipient-s-anti-tumor-immunity
#9
Ting Yan, Chen Luo, Xianmo Yang, Lei Ji, Shiqiao Luo
Objective To study the effect of short-term use of rapamycin(Rap) combined with regulatory T cells (Tregs) on the long-time survival of allogeneic mouse cardiac transplant, and its impact on the anti-tumor immunity of recipient. MethodsMouse Tregs were purified from recipients' spleen by magnetic activated cell sorting (MACS), and expanded by CD3/CD28 monoclonal antibody immunomagnetic beads and 2000 U/mL recombinant mouse IL-2 (rmIL-2) ex vivo. The purity was tested by fluorescence-activated cell sorting (FACS)...
February 2017: Xi Bao Yu Fen Zi Mian Yi Xue za Zhi, Chinese Journal of Cellular and Molecular Immunology
https://www.readbyqxmd.com/read/28068321/analysis-of-dna-methylation-in-single-circulating-tumor-cells
#10
C F Pixberg, K Raba, F Müller, B Behrens, E Honisch, D Niederacher, H Neubauer, T Fehm, W Goering, W A Schulz, P Flohr, G Boysen, M Lambros, J S De Bono, W T Knoefel, C Sproll, N H Stoecklein, R P L Neves
Direct analysis of circulating tumor cells (CTCs) can inform on molecular mechanisms underlying systemic spread. Here we investigated promoter methylation of three genes regulating epithelial-to-mesenchymal transition (EMT), a key mechanism enabling epithelial tumor cells to disseminate and metastasize. For this, we developed a single-cell protocol based on agarose-embedded bisulfite treatment, which allows investigating DNA methylation of multiple loci via a multiplex PCR (multiplexed-scAEBS). We established our assay for the simultaneous analysis of three EMT-associated genes miR-200c/141, miR-200b/a/429 and CDH1 in single cells...
January 9, 2017: Oncogene
https://www.readbyqxmd.com/read/28039186/mir-10b-5p-is-a-novel-th17-regulator-present-in-th17-cells-from-ankylosing-spondylitis
#11
L Chen, M H Al-Mossawi, A Ridley, T Sekine, A Hammitzsch, J de Wit, D Simone, H Shi, F Penkava, M Kurowska-Stolarska, I Pulyakhina, J C Knight, T J Kim, P Bowness
OBJECTIVE: To determine the microRNA (miR) signature in ankylosing spondylitis (AS) T helper (Th)17 cells. METHODS: Interleukin (IL)-17A-producing CD4+ T cells from patients with AS and healthy controls were FACS-sorted for miR sequencing and qPCR validation. miR-10b function was determined by miR mimic expression followed by cytokine measurement, transcriptome analysis, qPCR and luciferase assays. RESULTS: AS Th17 cells exhibited a miR signature characterised by upregulation of miR-155-5p, miR-210-3p and miR-10b...
December 30, 2016: Annals of the Rheumatic Diseases
https://www.readbyqxmd.com/read/28004528/minimal-residual-disease-monitoring-in-childhood-b-lymphoblastic-leukemia-with-t-12-21-p13-q22-etv6-runx1-concordant-results-using-quantitation-of-fusion-transcript-and-flow-cytometry
#12
S J Alm, C Engvall, J Asp, L Palmqvist, J Abrahamsson, L Fogelstrand
INTRODUCTION: The translocation t(12;21)(p13;q22) resulting in the fusion gene ETV6-RUNX1, is the most frequent gene fusion in childhood B lymphoblastic leukemia. In the Nordic Society of Paediatric Haematology and Oncology ALL-2008 treatment protocol, treatment stratification in B-lineage ALL is based on results of minimal residual disease (MRD) analysis with fluorescence-activated cell sorting (FACS). In this study, we determined whether RT-qPCR of the ETV6-RUNX1 fusion transcript can be a reliable alternative for MRD analysis...
April 2017: International Journal of Laboratory Hematology
https://www.readbyqxmd.com/read/28003307/pi3k-inhibition-reduces-mammary-tumor-growth-and-facilitates-antitumor-immunity-and-anti-pd1-responses
#13
Jiqing Sai, Philip Owens, Sergey V Novitskiy, Oriana E Hawkins, Anna E Vilgelm, Jinming Yang, Tammy Sobolik, Nicole Lavender, Andrew C Johnson, Colt McClain, Gregory D Ayers, Mark C Kelley, Melinda Sanders, Ingrid A Mayer, Harold L Moses, Mark Boothby, Ann Richmond
Purpose: Metastatic breast cancers continue to elude current therapeutic strategies, including those utilizing PI3K inhibitors. Given the prominent role of PI3Kα,β in tumor growth and PI3Kγ,δ in immune cell function, we sought to determine whether PI3K inhibition altered antitumor immunity.Experimental Design: The effect of PI3K inhibition on tumor growth, metastasis, and antitumor immune response was characterized in mouse models utilizing orthotopic implants of 4T1 or PyMT mammary tumors into syngeneic or PI3Kγ-null mice, and patient-derived breast cancer xenografts in humanized mice...
December 21, 2016: Clinical Cancer Research: An Official Journal of the American Association for Cancer Research
https://www.readbyqxmd.com/read/27995384/quantitative-imaging-by-pixel-based-contrast-enhanced-ultrasound-reveals-a-linear-relationship-between-synovial-vascular-perfusion-and-the-recruitment-of-pathogenic-il-17a-f-il-23-cd161-cd4-t-helper-cells-in-psoriatic-arthritis-joints
#14
Ugo Fiocco, Roberto Stramare, Veronica Martini, Alessandro Coran, Francesco Caso, Luisa Costa, Mara Felicetti, Gaia Rizzo, Matteo Tonietto, Anna Scanu, Francesca Oliviero, Bernd Raffeiner, Maristella Vezzù, Francesca Lunardi, Raffaele Scarpa, David Sacerdoti, Leopoldo Rubaltelli, Leonardo Punzi, Andrea Doria, Enrico Grisan
To develop quantitative imaging biomarkers of synovial tissue perfusion by pixel-based contrast-enhanced ultrasound (CEUS), we studied the relationship between CEUS synovial vascular perfusion and the frequencies of pathogenic T helper (Th)-17 cells in psoriatic arthritis (PsA) joints. Eight consecutive patients with PsA were enrolled in this study. Gray scale CEUS evaluation was performed on the same joint immediately after joint aspiration, by automatic assessment perfusion data, using a new quantification approach of pixel-based analysis and the gamma-variate model...
February 2017: Clinical Rheumatology
https://www.readbyqxmd.com/read/27976414/low-testosterone-alters-the-activity-of-mouse-prostate-stem-cells
#15
Ye Zhou, Ben Copeland, Maya Otto-Duessel, Miaoling He, Susan Markel, Tim W Synold, Jeremy O Jones
BACKGROUND: Low serum testosterone (low T) has been repeatedly linked to worse outcomes in men with newly diagnosed prostate cancer (PC). How low T contributes to these outcomes is unknown. Here we demonstrate that exposure to low T causes significant changes in the mouse prostate and prostate stem cells. METHODS: Mice were castrated and implanted with capsules to achieve castrate, normal, or sub-physiological levels of T. After 6 weeks of treatment, LC-MS/MS was used to quantify the levels of T and dihydrotestosterone (DHT) in serum and prostate tissue...
December 14, 2016: Prostate
https://www.readbyqxmd.com/read/27942419/immune-suppression-mediated-by-myeloid-and-lymphoid-derived-immune-cells-in-the-tumor-microenvironment-facilitates-progression-of-thyroid-cancers-driven-by-hras-g12v-and-pten-loss
#16
Lee Ann Jolly, Nicole Massoll, Aime T Franco
Thyroid cancer is the most common endocrine malignancy and is predicted to be the 4(th) most commonly diagnosed cancer by 2030. Approximately one-half of follicular thyroid carcinomas (FTC) contain genetic alterations in RAS family members. Furthermore, Cowden's disease, which is characterized by loss of PTEN, predisposes for the development of FTC in humans. We have shown that thyroid specific expression of Hras(G12V) at endogenous levels and Pten inactivation (Hras(G12V)/Pten(-/-)/TPO-cre mice) leads to the development of FTCs that closely recapitulate human disease, with complete penetrance at one year...
October 2016: Journal of Clinical & Cellular Immunology
https://www.readbyqxmd.com/read/27939246/knockout-of-p2y12-aggravates-experimental-autoimmune-encephalomyelitis-in-mice-via-increasing-of-il-23-production-and-th17-cell-differentiation-by-dendritic-cells
#17
Jiang Zhang, Zhenlong Li, Xuefei Hu, Qiong Su, Cong He, Jing Liu, Hua Ren, Min Qian, Junling Liu, Shufang Cui, Wenzheng Jiang
Experimental autoimmune encephalomyelitis (EAE), a common model of multiple sclerosis (MS), is mainly mediated by CD4(+) T cells with demyelination and neurodegeneration of central nervous system (CNS). The loss of P2Y12 receptor might be associated with the pathogenesis of MS/EAE, but its potential mechanism is still not clear. In this study, more severe EAE developed in P2Y12-knockout (P2Y12-KO) mice compared to WT mice. Knockout of P2Y12 increased expression of IL-17A in the sera and proportion of Th17 cells in spleen and CNS...
December 7, 2016: Brain, Behavior, and Immunity
https://www.readbyqxmd.com/read/27933701/ccr2-mediates-helicobacter-pylori-induced-immune-tolerance-and-contributes-to-mucosal-homeostasis
#18
Xia Sun, Min Zhang, Mohamad El-Zaatari, Gray B Huffnagle, John Y Kao
BACKGROUND: We previously demonstrated that H. pylori infection leads to increased induction of regulatory T cells in local and systemic immune compartments. Here, we investigate the role of CCR2 in the tolerogenic programing of dendritic cells in a mouse model of H. pylori infection. MATERIALS AND METHODS: CCR2 deficient (CCR2KO) mice and wild-type (Wt) mice infected with H. pylori SS1 strain were analyzed by qPCR and FACS analysis. In vitro, bone marrow-derived DC on day 6 from CCR2KO and Wt mice cocultured with or without H...
April 2017: Helicobacter
https://www.readbyqxmd.com/read/27852609/il-17-mediates-neutrophil-infiltration-and-renal-fibrosis-following-recovery-from-ischemia-reperfusion-compensatory-role-of-natural-killer-cells-in-athymic-rats
#19
Purvi Mehrotra, Jason A Collett, Seth D McKinney, Jackson Stevens, Carlie M Ivancic, David P Basile
T cells have been implicated in the pathogenesis of acute kidney injury (AKI) and its progression to chronic kidney disease (CKD). Previous studies suggest that Th17 cells participate during the AKI-to-CKD transition, and inhibition of T cell activity by mycophenolate mofetil (MMF) or losartan attenuates the development of fibrosis following AKI. We hypothesized that T cell-deficient rats may have reduced levels of IL-17 cytokine leading to decreased fibrosis following AKI. Renal ischemis-reperfusion (I/R) was performed on T cell-deficient athymic rats (Foxn1(rnu-/rnu-)) and control euthymic rats (Foxn1(rnu-/+)), and CKD progression was hastened by unilateral nephrectomy at day 33 and subsequent exposure to 4...
March 1, 2017: American Journal of Physiology. Renal Physiology
https://www.readbyqxmd.com/read/27828665/aerobic-training-modulates-t-cell-activation-in-elderly-women-with-knee-osteoarthritis
#20
W F Gomes, A C R Lacerda, G E A Brito-Melo, S F Fonseca, E Rocha-Vieira, A A O Leopoldino, M R Amorim, V A Mendonça
Osteoarthritis of the knee (kOA) is a disease that mainly affects the elderly and can lead to major physical and functional limitations. However, the specific effects of walking, particularly on the immune system, are unknown. Therefore, this study aimed to analyze the effect of 12 weeks of walking (3×/week) on the leukocyte profile and quality of life (QL) of elderly women with kOA. Sixteen women (age: 67±4 years, body mass index: 28.07±4.16 kg/m2) participated in a walking program. The variables were assessed before and after 12 weeks of training with a progressively longer duration (30-55 min) and higher intensity (72-82% of HRmax determined using a graded incremental treadmill test)...
2016: Brazilian Journal of Medical and Biological Research, Revista Brasileira de Pesquisas Médicas e Biológicas
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