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T cell facs analysis

Daniel Jons, Maria Kneider, Linda Fogelstrand, Anders Jeppsson, Stefan Jacobsson, Oluf Andersen
Contemporary evidence supports that MS immunopathology starts in the peripheral lymphatic system. However, the site and character of crucial initiating events are unknown. We examined subsets of the first stages of blood cells in the bone marrow of 9 MS patients and 11 neurologically healthy controls using FACS analysis. The proportion of natural killer T cells was lower (P=0.045) in the bone marrow of MS patients, but proportions of hematogenous stem cells, myeloblasts, and B cell precursor subsets in the bone marrow did not differ between MS patients and controls...
October 15, 2016: Journal of Neuroimmunology
Rabih Halwani, Asma Sultana Shaik, Elaref Ratemi, Sibtain Afzal, Rosan Kenana, Saleh Al-Muhsen, Achraf Al Faraj
Drug resistance and the harmful side effects accompanying the prolonged corticosteroid treatment of chronic pulmonary diseases prompted the development of more specific anti-inflammatory approaches. Several strategies aiming to block IL4Rα, the receptor for a key pro-inflammatory pathway, were investigated. However, their efficiency was limited, mostly due to the systemic or subcutaneous route of administrations. In this paper, we examined the ability of an intranasal treatment with biocompatible nanoparticles targeting IL4Rα to control lung inflammation in ovalbumin (OVA)-sensitized mice...
October 7, 2016: Experimental & Molecular Medicine
Kiran Kumar K M, M Naveen Kumar, Rajeshwari H Patil, Rashmi Nagesh, Shubha M Hegde, Kavya K, R L Babu, Govindarajan T Ramesh, S Chidananda Sharma
Cadmium (Cd) is one of the well-known highly toxic environmental and industrial pollutants. Cd first accumulates in the nucleus, and later interacts with zinc finger proteins of anti-apoptotic genes; inhibit the binding of transcriptional factors and transcription. However, the role of Cd in oxidative stress and apoptosis is less understood. Hence the present study was undertaken to unveil the mechanism of action. A549 cells were treated with or without Cd and cell viability was measured by MTT assay. Treatment of cells with Cd show reduced viability in a dose dependent manner with IC50 of 45µM concentration...
August 15, 2016: Toxicology Mechanisms and Methods
F Javier Rubio, Xuan Li, Qing-Rong Liu, Raffaello Cimbro, Bruce T Hope
The study of neuroplasticity and molecular alterations in learned behaviors is switching from the study of whole brain regions to the study of specific sets of sparsely distributed activated neurons called neuronal ensembles that mediate learned associations. Fluorescence Activated Cell Sorting (FACS) has recently been optimized for adult rat brain tissue and allowed isolation of activated neurons using antibodies against the neuronal marker NeuN and Fos protein, a marker of strongly activated neurons. Until now, Fos-expressing neurons and other cell types were isolated from fresh tissue, which entailed long processing days and allowed very limited numbers of brain samples to be assessed after lengthy and complex behavioral procedures...
2016: Journal of Visualized Experiments: JoVE
Jayson Hardcastle, Lisa Mills, Courtney S Malo, Fang Jin, Cheyne Kurokawa, Hirosha Geekiyanage, Mark Schroeder, Jann Sarkaria, Aaron J Johnson, Evanthia Galanis
BACKGROUND: Glioblastoma (GBM) is the most common primary malignant brain tumor and has a dismal prognosis. Measles virus (MV) therapy of GBM is a promising strategy due to preclinical efficacy, excellent clinical safety, and its ability to evoke antitumor pro-inflammatory responses. We hypothesized that combining anti- programmed cell death protein 1 (anti-PD-1) blockade and MV therapy can overcome immunosuppression and enhance immune effector cell responses against GBM, thus improving therapeutic outcome...
September 23, 2016: Neuro-oncology
Seungbum Kim, Gary Wang, Gilberto Lobaton, Eric Li, Tao Yang, Mohan Raizada
OBJECTIVE: Our previous studies have demonstrated that gut microbial dysbiosis is linked to high blood pressure in patients. This was associated with decreases in butyrate- and acetate- producing microbial populations. Thus, our objective in this study was to investigate the hypothesis that infusion of butyrate would impact dysbiosis, gut immunity and attenuate hypertension. DESIGN AND METHOD: C57B6 mice were divided into 4 groups; Saline infused, Angiotensin II (750ng/kg/min) infused, Ang II infused and butyrate treated (0...
September 2016: Journal of Hypertension
Gloria Gronowicz, Eric R Secor, John R Flynn, Liisa T Kuhn
OBJECTIVE: To assess the effect of human biofield therapy, an integrative medicine modality, on the development of tumors and metastasis, and immune function in a mouse breast cancer model. METHODS: Mice were injected with 66cl4 mammary carcinoma cells. In study one, mice received biofield therapy after cell injection. In study two, mice were treated by the biofield practitioner only prior to cell injection. Both studies had two control groups of mock biofield treatments and phosphate-buffered saline injection...
September 2016: Journal of Integrative Medicine
Valentina Perri, Elena Gianchecchi, Riccardo Scarpa, Mariella Valenzise, Maria Manuela Rosado, Ezio Giorda, Antonino Crinò, Marco Cappa, Susi Barollo, Silvia Garelli, Corrado Betterle, Alessandra Fierabracci
APECED is a T-cell mediated disease with increased frequencies of CD8+ effector and reduction of FoxP3+ T regulatory cells. Antibodies against affected organs and neutralizing to cytokines are found in the peripheral blood. The contribution of B cells to multiorgan autoimmunity in Aire-/- mice was reported opening perspectives on the utility of anti-B cell therapy. We aimed to analyse the B cell phenotype of APECED patients compared to age-matched controls. FACS analysis was conducted on PBMC in basal conditions and following CpG stimulation...
September 9, 2016: Immunobiology
Valentina E Schneeberger, Viola Allaj, Eric E Gardner, J T Poirier, Charles M Rudin
Patient-derived xenograft (PDX) mouse models are increasingly used for preclinical therapeutic testing of human cancer. A limitation in molecular and genetic characterization of PDX tumors is the presence of integral murine stroma. This is particularly problematic for genomic sequencing of PDX models. Rapid and dependable approaches for quantitating stromal content and purifying the malignant human component of these tumors are needed. We used a recently developed technique exploiting species-specific polymerase chain reaction (PCR) amplicon length (ssPAL) differences to define the fractional composition of murine and human DNA, which was proportional to the fractional composition of cells in a series of lung cancer PDX lines...
2016: PloS One
Shefalee K Bhavsar, Yogesh Singh, Piyush Sharma, Vishal Khairnar, Zohreh Hosseinzadeh, Shaqiu Zhang, Monica Palmada, Ivan Sabolic, Hermann Koepsell, Karl S Lang, Philipp A Lang, Florian Lang
BACKGROUND: Similar to tumor cells, activated T-lymphocytes generate ATP mainly by glycolytic degradation of glucose. Lymphocyte glucose uptake involves non-concentrative glucose carriers of the GLUT family. In contrast to GLUT isoforms, Na+-coupled glucose-carrier SGLT1 accumulates glucose against glucose gradients and is effective at low extracellular glucose concentrations. The present study explored expression and regulation of SGLT1 in activated murine splenic cytotoxic T cells (CTLs) and human Jurkat T cells...
2016: Cellular Physiology and Biochemistry
L Milena Beschel, Martin Leu, Sybille D Reichardt, Margret Rave-Fränk, Markus A Schirmer, Christine Stadelmann, Martin Canis, Hendrik A Wolff, Holger M Reichardt
Treatment of head and neck squamous cell carcinoma (HNSCC) by chemoradiotherapy (CRT) often results in high-grade acute organ toxicity (HGAOT). As these adverse effects impair the patients' quality of life and the feasibility of the planned therapy, we sought to analyze immunological parameters in tumor material and blood samples obtained from 48 HNSCC patients in order to assess the potential to predict the individual acute organ toxicity. T cells in the tumor stroma were enriched in patients developing HGAOT whereas levels of soluble factors in the plasma and gene expression in whole blood did not coincide with the occurrence of acute organ toxicity...
August 29, 2016: Oncotarget
Sara Palchetti, Daniela Pozzi, Cristina Marchini, Augusto Amici, Cristina Andreani, Caterina Bartolacci, Luca Digiacomo, Valentina Gambini, Francesco Cardarelli, Carmine Di Rienzo, Giovanna Peruzzi, Heinz Amenitsch, Rocco Palermo, Isabella Screpanti, Giulio Caracciolo
To date, efficiency upon non-viral DNA delivery remains low and this implies the existence of unidentified transfection barriers. Here we explore the mechanisms of action of multicomponent (MC) cationic liposome/DNA complexes (lipoplexes) by a combination of reporter technologies, dynamic light scattering (DLS), synchrotron small angle X-ray scattering (SAXS), fluorescence activated cell sorting (FACS) analysis and laser scanning confocal microscopy (LSCM) in live cells. Lipofectamine - the gold standard among transfection reagents - was used as a reference...
August 23, 2016: Nanomedicine: Nanotechnology, Biology, and Medicine
Khalil Ahmed, Betsy T Kren, Md Joynal Abedin, Rachel I Vogel, Daniel P Shaughnessy, Lucas Nacusi, Vicci L Korman, Yingming Li, Scott M Dehm, Cheryl L Zimmerman, Gloria A Niehans, Gretchen M Unger, Janeen H Trembley
CK2, a protein serine/threonine kinase, promotes cell proliferation and suppresses cell death. This essential-for-survival signal demonstrates elevated expression and activity in all cancers examined, and is considered an attractive target for cancer therapy. Here, we present data on the efficacy of a tenfibgen (TBG) coated nanocapsule which delivers its cargo of siRNA (siCK2) or single stranded RNA/DNA oligomers (RNAi-CK2) simultaneously targeting CK2α and α' catalytic subunits. Intravenous administration of TBG-siCK2 or TBG-RNAi-CK2 resulted in significant xenograft tumor reduction at low doses in PC3-LN4 and 22Rv1 models of prostate cancer...
August 20, 2016: Oncotarget
Stefanie Pektor, Nicole Bausbacher, Georg Otto, Laura Lawaczeck, Stephan Grabbe, Mathias Schreckenberger, Matthias Miederer
INTRODUCTION: High uptake of [(18)F]-2-fluorodeoxyglucose ([(18)F]FDG) by inflammatory cells is a frequent cause of false positive results in [(18)F]FDG-positron-emission tomography (PET) for cancer diagnostics. Similar to cancer cells, immune cells undergo significant increases in glucose utilization following activation, e.g., in infectious diseases or after vaccination during cancer therapy. The aim of this study was to quantify certain immune effects in vitro and in vivo by [(18)F]FDG-PET after stimulation with TLR ligands and specific antibodies...
July 14, 2016: Nuclear Medicine and Biology
Itsaso Montalbán Del Barrio, Cornelia Penski, Laura Schlahsa, Roland G Stein, Joachim Diessner, Achim Wöckel, Johannes Dietl, Manfred B Lutz, Michel Mittelbronn, Jörg Wischhusen, Sebastian F M Häusler
BACKGROUND: Ovarian cancer (OvCA) tissues show abundant expression of the ectonucleotidases CD39 and CD73 which generate immunomodulatory adenosine, thereby inhibiting cytotoxic lymphocytes. Little, however, is known about the effect of adenosine on myeloid cells. Considering that tumor associated macrophages (TAM) and myeloid-derived suppressor cells (MDSC) constitute up to 20 % of OvCA tissue, we investigated the effect of adenosine on myeloid cells and explored a possible contribution of myeloid cells to adenosine generation in vitro and ex vivo...
2016: Journal for Immunotherapy of Cancer
C Kitsou, L Lazaros, S Bellou, G Vartholomatos, P Sakaloglou, E Hatzi, S Markoula, K Zikopoulos, T Tzavaras, I Georgiou
Retroelement transcripts are present in male and female gametes, where they are typically regulated by methylation, noncoding RNAs and transcription factors. Such transcripts are required for occurrence of retrotransposition events, while failure of retrotransposition control may exert negative effects on cellular function and proliferation. In order to investigate the occurrence of retrotransposition events in mouse epididymal spermatozoa and to address the impact of uncontrolled retroelement RNA expression in early preimplantation embryos, we performed in vitro fertilization experiments using spermatozoa preincubated with plasmid vectors containing the human retroelements LINE-1, HERVK-10 or the mouse retroelement VL30, tagged with an enhanced green fluorescence (EGFP) gene-based cassette...
September 2016: Reproduction: the Official Journal of the Society for the Study of Fertility
Bhavya Balan Chandrika, Mathew Steephan, T R Santhosh Kumar, A Sabu, M Haridas
AIM: Aberrant human epidermal growth factor receptor-2 (HER2) expression and constitutive mutant activation of its tyrosine kinase domain account for tumor aggression and therapy resistance in many types of cancers with major share in breast cancer cases. HER2 specific treatment modalities still face challenges owing to the side effects and acquired resistance of available therapeutics. Recently, the anti-proliferative and pro-apoptotic potential of phytochemicals, especially of flavonoids have become increasingly appreciated as powerful chemo preventive agents...
September 1, 2016: Life Sciences
Judith Manz, Elke Rodríguez, Abdou ElSharawy, Eva-Maria Oesau, Britt-Sabina Petersen, Hansjörg Baurecht, Gabriele Mayr, Susanne Weber, Jürgen Harder, Eva Reischl, Agatha Schwarz, Natalija Novak, Andre Franke, Stephan Weidinger
Gene mapping studies have consistently identified a susceptibility locus for atopic dermatitis and other inflammatory diseases on chromosome 11q13.5, with the strongest association observed for a common variant located in an intergenic region between the two annotated genes chromosome 11 open reading frame 30 (C11orf30) and leucine rich repeat containing 32 (LRRC32). Using a targeted resequencing approach we identified low-frequency and rare missense mutations within the LRRC32 gene encoding the glycoprotein A repetitions predominantly (GARP), a receptor on activated regulatory T cells that binds latent TGFβ...
July 20, 2016: Journal of Investigative Dermatology
Young-Hee Kim, Thi-Anh-Thuy Tran, Hyun-Ju Lee, Sook-In Jung, Je-Jung Lee, Wool-Youl Jang, Kyung-Sub Moon, In-Young Kim, Shin Jung, Tae-Young Jung
We investigated the use of cytotoxic T-lymphocyte (CTL) epitopes in peptide immunotherapy for glioblastoma. Three peptides (ERBB2, BIRC5 and CD99) were selected based on their peptide-T2 cell binding affinities and combined in a multipeptide cocktail or a branched multipeptide synthesized with mini-polyethylene glycol spacers. Dendritic cells (DCs) pulsed with the multipeptide cocktail or branched multipeptide were compared based on their immunophenotype and cytokine secretion. FACS analysis of alpha-type 1 polarized dendritic cells (αDC1s) revealed that both groups highly expressed CD80, CD83 and CD86, indicating that both treatments efficiently generated mature αDC1s with the expected phenotype...
July 8, 2016: Oncotarget
Neda Mousavi Niri, Arash Memarnejadian, Younes Pilehvar-Soltanahmadi, Mohammadreza Agha Sadeghi, Mehdi Mahdavi, Nasim Kheshtchin, Samaneh Arab, Afshin Namdar, Farhad Jadidi, Nosratollah Zarghami, Jamshid Hajati
INTRODUCTION: The critical role of regulatory T (Treg) cells in dampening immune responses against tumor cells is apparent. Therefore, several methods have been introduced for eliminating Treg. Among them, inducing immune responses against Treg cells expressing Foxp3 transcription factor is a hopeful approach to decrease the frequency of Tregs. In current study, we used the chimeric FoxP3-Fc(IgG) fusion construct/protein to effectively stimulate the immune responses against Treg cells...
September 2016: Journal of Immunotherapy
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