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T cell facs analysis

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https://www.readbyqxmd.com/read/29024986/stromal-fibroblasts-from-perimenopausal-endometrium-exhibit-a-different-transcriptome-than-those-from-the-premenopausal-endometrium
#1
David W Erikson, Fatima Barragan, Terhi T Piltonen, Joseph C Chen, Shaina Balayan, Juan C Irwin, Linda C Giudice
Human endometrium undergoes extensive regeneration on a cyclic basis in premenopausal women and likely occurs through the contribution of stem/progenitor cells. Menopause results in the permanent cessation of menstrual cycles and is preceded by perimenopause, a period of several years in which endocrine and biological changes occur and is a period of risk for endometrial proliferative disorders. The objectives of this study were to identify mesenchymal stem cells (eMSC) and stromal fibroblasts (eSF) in endometrium of perimenopausal women and perform expression profile analysis of perimenopausal eMSC and eSF to gain insight into the biology of stem/progenitor and lineage cell populations during the transition to menopause...
August 19, 2017: Biology of Reproduction
https://www.readbyqxmd.com/read/29020964/il-27-mediates-hla-class-i-up-regulation-which-can-be-inhibited-by-the-il-6-pathway-in-hla-deficient-small-cell-lung-cancer-cells
#2
Grazia Carbotti, Amin Reza Nikpoor, Paola Vacca, Rosaria Gangemi, Chiara Giordano, Francesco Campelli, Silvano Ferrini, Marina Fabbi
BACKGROUND: Recently, immunotherapy with anti-PD-1 antibodies has shown clinical benefit in recurrent Small Cell Lung Cancer (SCLC). Since anti-PD-1 re-activates anti-tumor Cytotoxic T Lymphocyte (CTL) responses, it is crucial to understand the mechanisms regulating HLA class I, and PD-L1 expression in HLA-negative SCLC. Here we addressed the role of IL-27, a cytokine related to both IL-6 and IL-12 families. METHODS: The human SCLC cell lines NCI-N592, -H69, -H146, -H446 and -H82 were treated in vitro with different cytokines (IL-27, IFN-γ, IL-6 or a soluble IL-6R/IL-6 chimera [sIL-6R/IL-6]) at different time points and analyzed for tyrosine-phosphorylated STAT proteins by Western blot, for surface molecule expression by immunofluorescence and FACS analyses or for specific mRNA expression by QRT-PCR...
October 11, 2017: Journal of Experimental & Clinical Cancer Research: CR
https://www.readbyqxmd.com/read/29016555/interleukin-10-regulated-tumour-tolerance-in-non-small-cell-lung-cancer
#3
Julius Malte Vahl, Juliane Friedrich, Susanne Mittler, Sonja Trump, Lisanne Heim, Katerina Kachler, Liubov Balabko, Nicole Fuhrich, Carol-Immanuel Geppert, Denis Iulian Trufa, Nina Sopel, Ralf Rieker, Horia Sirbu, Susetta Finotto
BACKGROUND: Lung cancer is the most life-threatening cancer type worldwide. Treatment options include surgery, radio- and chemotherapy, as well as the use of immunomodulatory antibodies. Interleukin (IL)-10 is an immunosuppressive cytokine involved in tumour immune escape. METHODS: Immunohistochemistry (IHC) on human lung surgery tissue as well as human tumour cell line cultures, FACS analysis, real-time PCR and experimental lung cancer. RESULTS: Here we discovered a positive correlation between IL-10 and IL-10 receptor (IL-10R) expression in the lung with tumour diameter in patients with lung cancer (non-small cell lung cancer), the most life-threatening cancer type worldwide...
October 10, 2017: British Journal of Cancer
https://www.readbyqxmd.com/read/28951258/ibrutinib-suppresses-alloantibody-responses-in-a-mouse-model-of-allosensitization
#4
Irene Kim, Gordon Wu, Ning-Ning Chai, Andrew S Klein, Stanley Jordan
BACKGROUND: Ibrutinib is a Bruton's tyrosine Kinase (BTK) antagonist that inhibits B cell receptor (BCR) signaling. Complete BTK deficiency is associated with absence of B-cells. Ibrutinb is currently approved by FDA for treatment of B-cell malignancies, including Waldenström macroglobulinaemia. We recently carried out studies to determine if ibrutinib could modify alloantibody responses. MATERIALS AND METHODS: A mouse model of allogenic sensitization using a C57BL/6 mouse as the recipient of a skin allograft from an HLA-A2 transgenic mouse was utilized to examine the effects of ibrutinib on alloantibody responses and B cell effector functions...
September 23, 2017: Transplant Immunology
https://www.readbyqxmd.com/read/28951199/a-t-cell-specific-knockout-reveals-an-important-role-for-protease-activated-receptor-2-in-lymphocyte-development
#5
Nidhish Francis, Alison L Every, Babatunde A Ayodele, Robert N Pike, Eleanor J Mackie, Charles N Pagel
Activation of protease-activated receptor-2 (PAR2) expressed by T cells has been linked to the bone loss associated with periodontitis. We generated PAR2 conditional-null mice and crossed these with mice expressing Cre recombinase under control of the Lck proximal promoter, to produce T cell-specific PAR2-null mice in order to further study the cellular mechanism involved in periodontitis. Here we report that efficient deletion of PAR2 in thymocytes isolated from T cell-specific PAR2-null mice resulted in thymic and splenic hypoplasia and a reduction in the cells of the cortex and a loss of distinction between the cortex and the medulla of the thymus...
September 22, 2017: International Journal of Biochemistry & Cell Biology
https://www.readbyqxmd.com/read/28934507/a-comprehensive-cell-specific-microrna-catalogue-of-human-peripheral-blood
#6
Simonas Juzenas, Geetha Venkatesh, Matthias Hübenthal, Marc P Hoeppner, Zhipei Gracie Du, Maren Paulsen, Philip Rosenstiel, Philipp Senger, Martin Hofmann-Apitius, Andreas Keller, Limas Kupcinskas, Andre Franke, Georg Hemmrich-Stanisak
With this study, we provide a comprehensive reference dataset of detailed miRNA expression profiles from seven types of human peripheral blood cells (NK cells, B lymphocytes, cytotoxic T lymphocytes, T helper cells, monocytes, neutrophils and erythrocytes), serum, exosomes and whole blood. The peripheral blood cells from buffy coats were typed and sorted using FACS/MACS. The overall dataset was generated from 450 small RNA libraries using high-throughput sequencing. By employing a comprehensive bioinformatics and statistical analysis, we show that 3' trimming modifications as well as composition of 3' added non-templated nucleotides are distributed in a lineage-specific manner-the closer the hematopoietic progenitors are, the higher their similarities in sequence variation of the 3' end...
September 19, 2017: Nucleic Acids Research
https://www.readbyqxmd.com/read/28932631/tadalafil-has-biologic-activity-in-human-melanoma-results-of-a-pilot-trial-with-tadalafil-in-patients-with-metastatic-melanoma-tame
#7
Jessica C Hassel, Huanhuan Jiang, Carolin Bender, Julia Winkler, Alexandra Sevko, Ivan Shevchenko, Niels Halama, Antonia Dimitrakopoulou-Strauss, Walter E Haefeli, Dirk Jäger, Alexander Enk, Jochen Utikal, Viktor Umansky
Myeloid-derived suppressor cells (MDSCs) are known to play a critical role in the suppression of T cell antitumor responses. Our preclinical data showed that the phosphodiesterase (PDE)-5 inhibitor sildenafil impaired MDSC functions, enhanced intratumoral T cell activity and prolonged survival of melanoma-bearing mice. In this study, we evaluated biologic effects, safety and efficacy of palliative treatment with the PDE-5 inhibitor tadalafil in metastatic melanoma patients. We conducted an open-label, dose de-escalation trial with tadalafil in pretreated metastatic melanoma patients...
2017: Oncoimmunology
https://www.readbyqxmd.com/read/28904432/impact-of-steroids-on-the-inflammatory-response-after-ischemic-acute-kidney-injury-in-rats
#8
J Fontana, A Vogt, A Hohenstein, U Vettermann, E Doroshenko, E Lammer, B A Yard, S Hoeger
Inflammation plays a crucial role in acute kidney injury (AKI). The current study was designed to analyze the influence of prednisolone treatment on the inflammatory reaction during the first 96 h after AKI induction in a rat model. AKI was induced by unilateral clipping of the renal vessels. The treatment group received prednisolone 5 mg/kg s.c. daily. Infiltration rates of macrophages, leukocytes, and T-cells (24, 96 h) as well as plasma concentrations of the inflammatory markers intercellular adhesion molecule, interleukin-1 beta (IL-1β), IL-18, IL-6, and tumor necrosis factor-alpha (0, 6, 24, 96 h) were determined by fluorescence-activated cell sorting (FACS) analysis only...
September 2017: Indian Journal of Nephrology
https://www.readbyqxmd.com/read/28900857/endovascular-biopsy-in-vivo-cerebral-aneurysm-endothelial-cell-sampling-and-gene-expression-analysis
#9
Daniel L Cooke, David B McCoy, Van V Halbach, Steven W Hetts, Matthew R Amans, Christopher F Dowd, Randall T Higashida, Devon Lawson, Jeffrey Nelson, Chih-Yang Wang, Helen Kim, Zena Werb, Charles McCulloch, Tomoki Hashimoto, Hua Su, Zhengda Sun
There is limited data describing endothelial cell (EC) gene expression between aneurysms and arteries partly because of risks associated with surgical tissue collection. Endovascular biopsy (EB) is a lower risk alternative to conventional surgical methods, though no such efforts have been attempted for aneurysms. We sought (1) to establish the feasibility of EB to isolate viable ECs by fluorescence-activated cell sorting (FACS), (2) to characterize the differences in gene expression by anatomic location and rupture status using single-cell qPCR, and (3) to demonstrate the utility of unsupervised clustering algorithms to identify cell subpopulations...
September 13, 2017: Translational Stroke Research
https://www.readbyqxmd.com/read/28836203/isolation-of-live-fibroblasts-by-fluorescence-activated-cell-sorting
#10
Tripp Leavitt, Michael S Hu, Michael T Longaker
Flow cytometry is a powerful tool in cell biology in that it allows real-time characterization of cellular phenotypes. Additionally, through the process of fluorescence-activated cell sorting (FACS), living cells can be isolated for future in vitro experiments, including single cell analysis. Here we describe the isolation of live fibroblasts from the dermis of the skin in mice using FACS. This method circumvents the need for ex vivo expansion in cell culture, which can alter phenotypic and functional characteristics of cells...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28835380/single-dose-of-the-cxcr4-antagonist-bl-8040-induces-rapid-mobilization-for-the-collection-of-human-cd34-cells-in-healthy-volunteers
#11
Michal Abraham, Yaron Pereg, Baruch Bulvik, Shiri Klein, Inbal Mishalian, Hanna Wald, Orly Eizenberg, Katia Beider, Arnon Nagler, Rottem Golan, Abi Vainstein, Arnon Aharon, Eithan Galun, Yoseph Caraco, Reuven Or, Amnon Peled
PURPOSE: The potential of the high affinity CXCR4 antagonist BL-8040 as a monotherapy mobilizing agent and its derived graft composition and quality were evaluated in a phase I clinical study in healthy volunteers (NCT02073019). EXPERIMENTAL DESIGN: The first part of the study was a randomized, double-blind, placebo-controlled dose escalation phase. The second part of the study was an open label phase, in which 8 subjects received a single injection of BL-8040 (1mg/kg) and approximately 4hrs later underwent a standard leukapheresis procedure...
August 23, 2017: Clinical Cancer Research: An Official Journal of the American Association for Cancer Research
https://www.readbyqxmd.com/read/28758259/transient-receptor-potential-melastatin-4-channel-is-required-for-rat-dental-pulp-stem-cell-proliferation-and-survival
#12
T D Ngoc Tran, K E Stovall, T Suantawee, Y Hu, S Yao, L-J Yang, S Adisakwattana, H Cheng
OBJECTIVES: Investigate the role of the transient receptor potential melastatin 4 (TRPM4) channel in rat dental pulp stem cell (DPSC) proliferation and survival. MATERIALS AND METHODS: Immunofluorescence and FACS analysis were used to detect the stem cell marker CD90. Alizarin Red S and Oil Red O staining were used to identify osteoblast and adipocyte differentiation, respectively. To characterize TRPM4, patch-clamp recordings were obtained from single cells in the whole-cell configuration mode...
October 2017: Cell Proliferation
https://www.readbyqxmd.com/read/28710827/comparison-of-non-coding-rnas-in-exosomes-and-functional-efficacy-of-human-embryonic-stem-cell-versus-induced-pluripotent-stem-cell-derived-cardiomyocytes
#13
Won Hee Lee, Wen-Yi Chen, Ning-Yi Shao, Dan Xiao, Xulei Qin, Natalie Baker, Hye Ryeong Bae, Tzu-Tang Wei, Yongjun Wang, Praveen Shukla, Haodi Wu, Kazuki Kodo, Sang-Ging Ong, Joseph C Wu
Both human embryonic stem cell-derived cardiomyocytes (ESC-CMs) and human induced pluripotent stem cell-derived CMs (iPSC-CMs) can serve as unlimited cell sources for cardiac regenerative therapy. However, the functional equivalency between human ESC-CMs and iPSC-CMs for cardiac regenerative therapy has not been demonstrated. Here, we performed a head-to-head comparison of ESC-CMs and iPSC-CMs in their ability to restore cardiac function in a rat myocardial infarction (MI) model as well as their exosomal secretome...
July 14, 2017: Stem Cells
https://www.readbyqxmd.com/read/28706011/increased-ifn%C3%AE-t-cells-are-responsible-for-the-clinical-responses-of-low-dose-dna-demethylating-agent-decitabine-antitumor-therapy
#14
Xiang Li, Yan Zhang, Meixia Chen, Qian Mei, Yang Liu, Kaichao Feng, Hejin Jia, Liang Dong, Lu Shi, Lin Liu, Jing Nie, Weidong Han
Purpose: Low-dose DNA-demethylating agent decitabine therapy is effective in a subgroup of cancer patients. It remains largely elusive for the biomarker to predict therapeutic response and the underlying antitumor mechanisms, especially the impact on host antitumor immunity.Experimental Design: The influence of low-dose decitabine on T cells was detected both in vitro and in vivo Moreover, a test cohort and a validation cohort of advanced solid tumor patients with low-dose decitabine-based treatment were involved...
October 15, 2017: Clinical Cancer Research: An Official Journal of the American Association for Cancer Research
https://www.readbyqxmd.com/read/28702612/rapid-label-free-cd4-testing-using-a-smartphone-compatible-device
#15
Manoj Kumar Kanakasabapathy, Hardik J Pandya, Mohamed Shehata Draz, Manjyot Kaur Chug, Magesh Sadasivam, Shreya Kumar, Behzad Etemad, Vinish Yogesh, Mohammadali Safavieh, Waseem Asghar, Jonathan Z Li, Athe M Tsibris, Daniel R Kuritzkes, Hadi Shafiee
The most recent guidelines have called for a significant shift towards viral load testing for HIV/AIDS management in developing countries; however point-of-care (POC) CD4 testing still remains an important component of disease staging in multiple developing countries. Advancements in micro/nanotechnologies and consumer electronics have paved the way for mobile healthcare technologies and the development of POC smartphone-based diagnostic assays for disease detection and treatment monitoring. Here, we report a simple, rapid (30 minutes) smartphone-based microfluidic chip for automated CD4 testing using a small volume (30 μL) of whole blood...
August 22, 2017: Lab on a Chip
https://www.readbyqxmd.com/read/28683082/the-herbal-decoction-modified-danggui-buxue-tang-attenuates-immune-mediated-bone-marrow-failure-by-regulating-the-differentiation-of-t-lymphocytes-in-an-immune-induced-aplastic-anemia-mouse-model
#16
Peiying Deng, Xue Li, Yi Wei, Juan Liu, Meng Chen, Yamei Xu, Bin Dong, Lingqun Zhu, Limin Chai
Angelicae Sinensis, Radix Astragali and Rhizoma Coptidis are all herbs of modified Danggui Buxue Tang (DGBX) and are extensively applied herbs in traditional Chinese medicine for the treatment of anemia and inflammation. In this study, immune-induced AA mice were used as an animal model, and the immunosuppressive agent, Ciclosporin A (CsA), was used as a positive control. Multiple pro-inflammatory cytokines were examined by bead-based multiplex flow cytometry. The T-cell subsets were assessed using a fluorescence-activated cell sorter (FACS)...
2017: PloS One
https://www.readbyqxmd.com/read/28682940/monocyte-dependent-suppression-of-t-cell-function-in-postoperative-patients-and-abdominal-sepsis
#17
Markus Albertsmeier, Niclas J Prix, Hauke Winter, Alexandr Bazhin, Jens Werner, Martin K Angele
INTRODUCTION: Surgical trauma causes inflammation and postoperative immunosuppression. Previous studies have shown a T-cell dependent suppression of MHC II expression and other functions of antigen presenting cells. Aim of this study was to determine which immune cell initiates postoperative immunosuppression and consecutive sepsis. METHODS: We separated T-cells and monocytes in human abdominal surgery (n = 11) patients preoperatively as well as 24 hrs postoperatively and in patients who developed postoperative sepsis (n = 6)...
July 4, 2017: Shock
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
#18
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...
September 2017: Journal of Leukocyte Biology
https://www.readbyqxmd.com/read/28595132/functionality-based-method-for-simultaneous-isolation-of-rodent-hepatic-sinusoidal-cells
#19
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
#20
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 pancreatic polypeptide cells but not to α- or δ-cells. RNA-sequencing analysis of FACS-purified Abcc8(-/-) β-cells confirmed an increase in Ppy gene expression and revealed altered expression of more than 4,200 genes, many of which are involved in Ca(2+) signaling, the maintenance of β-cell identity, and cell adhesion...
August 2017: Diabetes
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