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T cell facs analysis

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https://www.readbyqxmd.com/read/28637895/cd4-cd8%C3%AE-double-positive-t-cells-in-skin-draining-lymph-nodes-respond-to-inflammatory-signals-from-the-skin
#1
Nana H Overgaard, Jazmina L Cruz, Jennifer A Bridge, Hendrik J Nel, Ian H Frazer, Nicole L La Gruta, Antje Blumenthal, Raymond J Steptoe, James W Wells
CD4(+)CD8(+) double-positive (DP), mature, peripheral T cells are readily detectable in a variety of species and tissues. Despite a common association with autoimmune and malignant skin disorders, however, little is understood about their role or function. Herein, we show that DP T cells are readily detectable in the blood, spleen, and peripheral lymph nodes of naïve C57BL/6 mice. DP T cells were also present in Jα18(-/-) and CD1d(-/-) mice, indicating that these cells are not NK-T cells. After skin administration of CASAC adjuvant, but not Quil A adjuvant, both total DP T cells and skin-infiltrating DP T cells increased in number...
June 21, 2017: Journal of Leukocyte Biology
https://www.readbyqxmd.com/read/28595132/functionality-based-method-for-simultaneous-isolation-of-rodent-hepatic-sinusoidal-cells
#2
L Stradiot, S Verhulst, T Roosens, C I Øie, I M Moya, G Halder, I Mannaerts, L A van Grunsven
Chronic liver disease is the result of long term exposure to viruses or toxins such as alcohol, fat and drugs, and forms the basis for the development of liver fibrosis and primary liver cancer. In vitro and in vivo models are key to study the pathways involved in chronic liver disease and for the development of therapeutics. 3D co-culture systems are becoming the in vitro standard, which requires freshly isolated primary hepatic cells. We developed a novel isolation method to simultaneously isolate liver sinusoidal endothelial cells (LSECs), Kupffer cells (KCs) and hepatic stellate cells (HSCs)...
September 2017: Biomaterials
https://www.readbyqxmd.com/read/28550109/chronic-%C3%AE-cell-depolarization-impairs-%C3%AE-cell-identity-by-disrupting-a-network-of-ca-2-regulated-genes
#3
Jennifer S Stancill, Jean-Philippe Cartailler, Hannah W Clayton, James T O'Connor, Matthew T Dickerson, Prasanna K Dadi, Anna B Osipovich, David A Jacobson, Mark A Magnuson
We used mice lacking Abcc8, a key component of the β-cell KATP-channel, to analyze the effects of a sustained elevation in the intracellular Ca(2+) concentration ([Ca(2+)]i) on β-cell identity and gene expression. Lineage tracing analysis revealed the conversion of β-cells lacking Abcc8 into PP-cells, but not to α- or δ-cells. RNA-Seq analysis of FACS-purified Abcc8(-/-) β-cells confirmed an increase in Ppy gene expression, and revealed altered expression of over 4,200 genes, many of which are involved in Ca(2+)-signaling, the maintenance of β-cell identity, and cell adhesion...
May 26, 2017: Diabetes
https://www.readbyqxmd.com/read/28548949/dietary-grape-seed-proanthocyanidins-inactivate-regulatory-t-cells-by-promoting-ner-dependent-dna-repair-in-dendritic-cells-in-uvb-exposed-skin
#4
Mudit Vaid, Ram Prasad, Tripti Singh, Santosh K Katiyar
Ultraviolet B (UVB) radiation induces regulatory T cells (Treg cells) and depletion of these Treg cells alleviates immunosuppression and inhibits photocarcinogenesis in mice. Here, we determined the effects of dietary grape seed proanthocyanidins (GSPs) on the development and activity of UVB-induced Treg cells. C3H/HeN mice fed a GSPs (0.5%, w/w)-supplemented or control diet were exposed to UVB (150 mJ/cm2) radiation, sensitized to 2,4-dinitrofluorobenzene (DNFB) and sacrificed 5 days later. FACS analysis indicated that dietary GSPs decrease the numbers of UVB-induced Treg cells...
May 15, 2017: Oncotarget
https://www.readbyqxmd.com/read/28542359/performance-evaluation-of-bd-facspresto%C3%A2-point-of-care-cd4-analyzer-to-enumerate-cd4-counts-for-monitoring-hiv-infected-individuals-in-nigeria
#5
Olubunmi Ruth Negedu-Momoh, Feyisayo Ebenezer Jegede, Ali Yakubu, Oluseyi Balogun, Musa Abdullahi, Titilope Badru, Edward Adekola Oladele, Chinedu Agbakwuru, Hadiza Khamofu, Kwasi Torpey
BACKGROUND: Despite the upsurge in support and intervention of donor agencies in HIV care and treatment programing in Sub-Sahara African, antiretroviral (ART) programs are still confronted with access and coverage challenges which influence enrolment of new patients. This study investigated the validity of point of care BD FACSPresto™ CD4 analyzer for CD4+ cell count, overall agreement, correlation, sensitivity, and specificity in comparison to a reference standard flow cytometry method...
2017: PloS One
https://www.readbyqxmd.com/read/28508475/functional-properties-of-peripheral-cd8-%C3%A2-t-cells-in-patients-with-repeated-implantation-failure
#6
Biao Yin, Yong Zeng, Tonghua Wu, Shuyi Yu, Jian Xu, Su Liu, Lianghui Diao, Zhenfu Zhao, Desheng Liang, Yuye Li
CD8(+) T cells are the main candidates to recognize and respond to fetal HLA-C at the fetal-maternal interface, but data on the amount of peripheral CD8(+) T cells and their functions during the window of implantation in recurrent implantation failure (RIF) patients are limited. Peripheral blood was obtained from 56 women with RIF and 16 fertile women in the mid-luteal phase of the menstrual cycle, and the CD8(+) T cells were determined by FACS analysis. No statistical differences in the proportion of peripheral CD8(+) T cells were observed among the women with RIF and the control group...
May 15, 2017: American Journal of Reproductive Immunology: AJRI
https://www.readbyqxmd.com/read/28475971/novel-mononuclear-cu-ii-terpyridine-complexes-impact-of-fused-ring%C3%A2-thiophene-and-thiazole-head-groups-towards-dna-bsa-interaction-%C3%A2-cleavage-and-antiproliferative-activity-on-hepg2-and%C3%A2-triple%C3%A2-negative-cal-51%C3%A2-cell-line
#7
V M Manikandamathavan, M Thangaraj, T Weyhermuller, R P Parameswari, V Punitha, N Narasimha Murthy, Balachandran Unni Nair
Two mononuclear copper (II) terpyridine complexes namely, [Cu(Btptpy) (ClO4)](ClO4) 1, and [Cu(Bttpy) (ClO4)](ClO4) 2, (Btptpy (L1) = 4'-(Benzothiophene)-2,2':6',2″-terpyridine, Bttpy (L2) = 4'-(Benzylthiazolyl)-2,2':6',2″-terpyridine) have been synthesized and characterized. Single crystal X-ray diffraction shows that, both ligands belong to monoclinic crystal system with space group P21/c (L1) and P21/n (L2). Absorption spectral titration, DNA melting study, circular dichroism and viscosity measurement reveal that, complex 1 and 2 bind with DNA through intercalation...
April 15, 2017: European Journal of Medicinal Chemistry
https://www.readbyqxmd.com/read/28465541/transcriptional-landscape-of-epithelial-and-immune-cell-populations-revealed-through-facs-seq-of-healthy-human-skin
#8
Richard S Ahn, Keyon Taravati, Kevin Lai, Kristina M Lee, Joanne Nititham, Rashmi Gupta, David S Chang, Sarah T Arron, Michael Rosenblum, Wilson Liao
Human skin consists of multiple cell types, including epithelial, immune, and stromal cells. Transcriptomic analyses have previously been performed from bulk skin samples or from epithelial and immune cells expanded in cell culture. However, transcriptomic analysis of bulk skin tends to drown out expression signals from relatively rare cells while cell culture methods may significantly alter cellular phenotypes and gene expression profiles. To identify distinct transcriptomic profiles of multiple cell populations without substantially altering cell phenotypes, we employed a fluorescence activated cell sorting method to isolate keratinocytes, dendritic cells, CD4+ T effector cells, and CD8+ T effector cells from healthy skin samples, followed by RNA-seq of each cell population...
May 2, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28464475/isolation-of-cd248-expressing-stromal-vascular-fraction-for-targeted-improvement-of-wound-healing
#9
Elizabeth Brett, Elizabeth R Zielins, Monica Chin, Michael Januszyk, Charles P Blackshear, Michael Findlay, Arash Momeni, Geoffrey C Gurtner, Michael T Longaker, Derrick C Wan
Wound healing remains a global issue of disability, cost, and health. Addition of cells from the stromal vascular fraction (SVF) of adipose tissue has been shown to increase the rate of full thickness wound closure. This study aimed to investigate the angiogenic mechanisms of CD248+ SVF cells in the context of full thickness excisional wounds. Single cell transcriptional analysis was used to identify and cluster angiogenic gene-expressing cells, which was then correlated with surface marker expression. SVF cells isolated from human lipoaspirate were FACS sorted based on the presence of CD248...
May 2, 2017: Wound Repair and Regeneration
https://www.readbyqxmd.com/read/28417335/poor-prognosis-of-patients-with-triple-negative-breast-cancer-can-be-stratified-by-rank-and-rankl-dual-expression
#10
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
#11
Qin Lu, Yinan Liu, Yang Wang, Weiping Wang, Zhe Yang, Tao Li, Yuyao Tian, Ping Chen, Kangtao Ma, Zhuqing Jia, Chunyan Zhou
To investigate the effects of rapamycin on cardiac differentiation, murine embryonic stem cells (ESCs) were induced into cardiomyocytes by 10(-4) M ascorbic acid (AA), 20 nM rapamycin alone or 0.01% solvent DMSO. We found that rapamycin alone was insufficient to initiate cardiomyogenesis. Then, the ESCs were treated with AA and rapamycin (20 nM) or AA and DMSO (0.01%) as a control. Compared with control, mouse ESCs (mESCs) treated with rapamycin (20 nM) and AA yielded a significantly higher percentage of cardiomyocytes, as confirmed by the percentage of beating embryonic bodies (EBs), the immunofluorescence and FACS analysis...
June 30, 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
#12
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
#13
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...
June 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
#14
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
#15
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...
August 2017: Redox Biology
https://www.readbyqxmd.com/read/28202731/microfluidic-droplet-platform-for-ultrahigh-throughput-single-cell-screening-of-biodiversity
#16
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
#17
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
#18
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
#19
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...
June 8, 2017: Oncogene
https://www.readbyqxmd.com/read/28039186/mir-10b-5p-is-a-novel-th17-regulator-present-in-th17-cells-from-ankylosing-spondylitis
#20
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...
March 2017: Annals of the Rheumatic Diseases
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