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Immunophenotyping by flow cytometry

Michael R Pranzatelli, Tyler J Allison, Elizabeth D Tate
INTRODUCTION: Flow cytometric cerebrospinal fluid (CSF) lymphocyte subset analysis has improved the diagnosis of neuroinflammation and identified multiple markers of inflammation in opsoclonus-myoclonus syndrome (OMS). The aim of this exploratory, retrospective study was to analyze the effect of immunotherapy on these markers to determine which agents are disease modifying. METHODS: Cross-sectional immunological observations were made in an IRB-approved case-control study, and patients were treated empirically...
March 5, 2018: European Journal of Paediatric Neurology: EJPN
Jacqueline Cloos, Jeffrey R Harris, Jeroen J W M Janssen, Angele Kelder, F Huang, Gerrit Sijm, Maike Vonk, Alexander N Snel, Jennifer R Scheick, Willemijn J Scholten, Jannemieke Carbaat-Ham, Dennis Veldhuizen, Diana Hanekamp, Yvonne J M Oussoren-Brockhoff, Gertjan J L Kaspers, Gerrit J Schuurhuis, A Kate Sasser, Gert Ossenkoppele
Response criteria in acute myeloid leukemia (AML) has recently been re-established, with morphologic examination utilized to determine whether patients have achieved complete remission (CR). Approximately half of the adult patients who entered CR will relapse within 12 months due to the outgrowth of residual AML cells in the bone marrow. The quantitation of these remaining leukemia cells, known as minimal or measurable residual disease (MRD), can be a robust biomarker for the prediction of these relapses. Moreover, retrospective analysis of several studies has shown that the presence of MRD in the bone marrow of AML patients correlates with poor survival...
March 5, 2018: Journal of Visualized Experiments: JoVE
Qingkai Dai, Xiaojuan Liu, Hui Yang, Siqi Guo, Yuefang Wang, Luyun Peng, Lei Ye, Lan Chen, Chunqi Lai, Qi Chen, Ge Zhang, Yongmei Jiang
Detection of aberrant antigen expression in acute lymphoblastic leukemia (ALL) by flow cytometric is proposed for the quantification of minimal residual disease (MRD). There are few studies that investigate the stability of the antigen expression in children with B lineage ALL at the end of remission induction therapy and determine its prognostic impact. Between 2010 and 2015, 691 bone marrow specimens of childhood ALL were sent at diagnosis for immunophenotypic characterization, and follow-up samples for MRD were analyzed on day 33...
March 2, 2018: Leukemia Research
Wei Duan, Cong Chen, Masudul Haque, Daniel Hayes, Mandi J Lopez
BACKGROUND: Use of bioscaffolds to direct osteogenic differentiation of adult multipotent stromal cells (MSCs) without exogenous proteins is a contemporary approach to bone regeneration. Identification of in vivo osteogenic contributions of exogenous MSCs on bioscaffolds after long-term implantation is vital to understanding cell persistence and effect duration. METHODS: This study was designed to quantify in vivo equine MSC osteogenesis on synthetic polymer scaffolds with distinct mineral combinations 9 weeks after implantation in a murine model...
March 9, 2018: Stem Cell Research & Therapy
Esteban Arrieta-Bolaños, Pietro Crivello, Maximilian Metzing, Thuja Meurer, Müberra Ahci, Julie Rytlewski, Marissa Vignali, Erik Yusko, Peter van Balen, Peter A Horn, J H Frederik Falkenburg, Katharina Fleischhauer
T cell alloreactivity is mediated by a self-human leukocyte antigen (HLA)-restricted T cell receptor (TCR) repertoire able to recognize both structurally similar and dissimilar allogeneic HLA molecules (i.e., differing by a single or several amino acids in their peptide-binding groove). We hypothesized that thymic selection on self-HLA molecules could have an indirect impact on the size and diversity of the alloreactive response. To test this possibility, we used TCR Vβ immunophenotyping and immunosequencing technology in a model of alloreactivity between self-HLA selected T cells and allogeneic HLA-DPB1 (DPB1) differing from self-DPB1*04:02 by a single (DPB1*02:01) or several (DPB1*09:01) amino acids in the peptide-binding groove...
2018: Frontiers in Immunology
Hailian Wang, Wei Ge, Yong Zhuang, Jinqiu Fu, Dong Li, Xiuli Ju
Background: Cord blood transplantation (CBT) can be a life-saving procedure in the treatment of a broad variety of disorders, including hematologic, immune, and genetic diseases. However, delayed platelet recovery hinders the application of CBT. Purpose: The aim of this study was to determine the optimal combination of cytokines to amplify megakaryocyte (Mk). Methods: CB CD34+ cells were obtained by immunomagnetic isolation and amplified under four different cytokine combinations...
January 2018: Journal of Cancer Research and Therapeutics
Gabriela Gainaru, Antonios Papadopoulos, Iraklis Tsangaris, Malvina Lada, Evangelos J Giamarellos-Bourboulis, Aikaterini Pistiki
BACKGROUND: Evidence on the changes in the absolute counts of monocyte subpopulations in sepsis is missing. METHODS: Firstly, absolute counts of circulating CD14pos /HLA-DRpos /CD45pos monocytes were measured by flow cytometry in 70 patients with Gram-negative sepsis and in 10 healthy volunteers. In the second phase, immunophenotyping was performed and the absolute count of circulating inflammatory monocytes and of circulating CD14dim /CD16pos /CD45pos patrolling monocytes were measured in another 55 patients and 10 healthy volunteers...
March 3, 2018: Critical Care: the Official Journal of the Critical Care Forum
Francesco Buccisano, Luca Maurillo, Maria Ilaria Del Principe, Ambra Di Veroli, Eleonora De Bellis, Annalisa Biagi, Anangiulia Zizzari, Valentina Rossi, Vito Rapisarda, Sergio Amadori, Maria Teresa Voso, Francesco Lo-Coco, William Arcese, Adriano Venditti
Response to therapy is affected by the genetic heterogeneity of acute myeloid leukemia (AML) and persistence of leukemic cells below the threshold of morphological complete remission (mCR). Such persistence is called minimal (or measurable) residual disease (MRD). Areas covered: MRD assessment allows early identification of patients who are at high risk of relapse and who should timely receive aggressive therapy (e.g. allogeneic stem cell transplantation) and of those with a good quality mCR in whom an aggressive front-line therapy can be spared, avoiding the harm of excessive treatment toxicity...
March 2, 2018: Expert Review of Hematology
Jinge Xu, Bin Liu, Shoubao Ma, Jubin Zhang, Yuhan Ji, Liangjing Xu, Mingqing Zhu, Suning Chen, Xiaojin Wu, Depei Wu
BACKGROUND/AIMS: Carcinoembryonic antigen-related cell adhesion molecule 1 (CEACAM1), also known as CD66a, is a member of the immunoglobulin (Ig) superfamily that belongs to the carcinoembryonic antigen (CEA) family which plays a dual role in cancer. Previous studies showed high expression of CEACAM1 in multiple myeloma (MM). The aim of this study was to investigate the biological consequences of CEACAM1 overexpression in MM. METHODS: pEGFP-N1-CEACAM1 and pcDNA3...
February 21, 2018: Cellular Physiology and Biochemistry
Michael R Pranzatelli, Tyler J Allison, Nathan R McGee, Elizabeth D Tate
Studies of cerebrospinal fluid (CSF) γδ T cells in children are limited, especially due to the lack of control data. In adults, gamma/delta T cells (TCR-γδ) residing in the intrathecal space are sometimes involved in neuroinflammation. To evaluate the possible role of γδ T cells in pediatric neuroinflammation, we immunophenotyped CSF and blood lymphocytes using flow cytometry in a case-control study of 100 children with non-inflammatory neurologic disorders (NIND), 312 with opsoclonus-myoclonus (OMS), and 23 with other inflammatory neurologic disorders (OIND)...
February 27, 2018: Clinical and Experimental Immunology
R Solís-Martínez, M Cancino-Marentes, G Hernández-Flores, P Ortiz-Lazareno, G Mandujano-Álvarez, C Cruz-Gálvez, E Sierra-Díaz, C Rodríguez-Padilla, L F Jave-Suárez, A Aguilar-Lemarroy, A Bravo-Cuellar
BACKGROUND: Transcription factor STAT3 has a prominent innate immunity effect on cancer progression. We determined the regulation of STAT3 in the immunophenotype modulation of macrophages from M1 into M2 induced by the cell-culture supernatant of the Prostate-Cancer line PC3. METHODS: Monocytes-macrophages from healthy donors were cultured in the supernatant of PC3 cells, membrane proteins, and intracytoplasmic and phosphorylated STAT3 were measured using flow cytometry, while cytokines and growth factors were studied using luminescence...
February 23, 2018: Immunology Letters
L Tauzin, V Campos, M Tichet
Reticulocytes and erythrocytes are the ultimate differentiated stages of erythropoiesis. In addition to being anucleate cells, they are characterized by the clearance of their mitochondrial pool or lack thereof. Given that for most research-oriented flow cytometry experiments erythrocytes and reticulocytes are often undesirable cell types, their identification and exclusion from analyses can be essential. Here, we describe a flow cytometric method based on cellular NAD(P)H-related autofluorescence, whose localization is mainly associated with mitochondria...
February 26, 2018: Cytometry. Part A: the Journal of the International Society for Analytical Cytology
Mariangeles Auat, Chandra Chiappin Cardoso, Iris Mattos Santos-Pirath, Renata Cristina Messores Rudolf-Oliveira, Camila Matiollo, Bárbara Gil Lange, Jessica Pires da Silva, Gisele Cristina Dametto, Mayara Marin Pirolli, Maria Daniela Holthausen Perico Colombo, Maria Claudia Santos-Silva
BACKGROUND: Flow cytometric immunophenotyping is deemed a fundamental tool for the diagnosis of B-cell neoplasms. Currently, the investigation of novel immunophenotypic markers has gained importance, as they can assist in the precise subclassification of B-cell malignancies by flow cytometry. Therefore, the purpose of the present study was to evaluate the expression of CD307a during normal B-cell maturation and in B-cell malignancies as well as to investigate its potential role in the differential diagnosis of these entities...
February 24, 2018: Cytometry. Part B, Clinical Cytometry
Etienne Patin, Milena Hasan, Jacob Bergstedt, Vincent Rouilly, Valentina Libri, Alejandra Urrutia, Cécile Alanio, Petar Scepanovic, Christian Hammer, Friederike Jönsson, Benoît Beitz, Hélène Quach, Yoong Wearn Lim, Julie Hunkapiller, Magge Zepeda, Cherie Green, Barbara Piasecka, Claire Leloup, Lars Rogge, François Huetz, Isabelle Peguillet, Olivier Lantz, Magnus Fontes, James P Santo, Stéphanie Thomas, Jacques Fellay, Darragh Duffy, Lluís Quintana-Murci, Matthew L Albert
The quantification and characterization of circulating immune cells provide key indicators of human health and disease. To identify the relative effects of environmental and genetic factors on variation in the parameters of innate and adaptive immune cells in homeostatic conditions, we combined standardized flow cytometry of blood leukocytes and genome-wide DNA genotyping of 1,000 healthy, unrelated people of Western European ancestry. We found that smoking, together with age, sex and latent infection with cytomegalovirus, were the main non-genetic factors that affected variation in parameters of human immune cells...
February 23, 2018: Nature Immunology
Simon Degandt, Bart Peeters, Stijn Jughmans, Nancy Boeckx, Xavier Bossuyt
BACKGROUND: Quantitation of lymphocyte subsets (B cells, T cells, CD4 and CD8 T cells and NK cells) classically relies on quantitation of lymphocytes and immunophenotyping by flow cytometry. AQUIOS CL (Beckman Coulter) is a fully automated system that performs an onboard volumetric cell count, automatically processes the sample (staining, lysing and fixation) and analyzes the results. We compared AQUIOS CL to a dual-platform analysis and evaluated analytical performance. METHODS: We evaluated precision, sample stability, inter-sample carryover, linearity and interpanel consistency...
February 21, 2018: Clinical Chemistry and Laboratory Medicine: CCLM
Malte Mohme, Simon Schliffke, Cecile L Maire, Alessandra Rünger, Laura Glau, Klaus C Mende, Jakob Matschke, Christina Gehbauer, Nuray Akyüz, Svenja Zapf, Mareike Holz, Miriam Schaper, Tobias Martens, Nils O Schmidt, Sven Peine, Manfred Westphal, Mascha Binder, Eva Tolosa, Katrin Lamszus
PURPOSE: Immunotherapeutic treatment strategies for glioblastoma (GBM) are under investigation in clinical trials. However, our understanding of the immune phenotype of GBM-infiltrating T-cells (TILs) and changes during disease progression is limited. Deeper insight is urgently needed to therapeutically overcome tumor-induced immune exhaustion. EXPERIMENTAL DESIGN: We used flow-cytometry and cytokine assays to profile TILs and blood lymphocytes (PBL) from GBM patients, comparing newly diagnosed or recurrent GBM to long-term survivors (LTS) and healthy donors...
February 14, 2018: Clinical Cancer Research: An Official Journal of the American Association for Cancer Research
Hossein Ranjbaran, Saeid Abediankenari, Masoud Mohammadi, Narjes Jafari, Alireza Khalilian, Zahra Rahmani, Marzei Momeninezhad Amiri, Pirooz Ebrahimi
Wharton`s jelly-derived mesenchymal stem cells (WJ-MSCs), have a high proliferation valency and they do not produce teratogen or carcinogen after subsequent transplantation. They are known as regenerative medicine. Thus more research is needed on the isolation and characterization of mesenchymal stem cells. In this experimental study, we obtained Wharton's jelly tissues from mothers during normal vaginal delivery, after obtaining their informed consent. Mesenchymal stem cells were isolated from cultured Wharton`s jelly, cultured, and were then examined for their proliferation, immunophenotypes, and differentiation capacities...
January 2018: Acta Medica Iranica
S Comazzi, M Cozzi, S Bernardi, D R Zanella, L Aresu, D Stefanello, L Marconato, V Martini
Flow cytometry (FC) is increasingly being used for immunophenotyping and staging of canine lymphoma. The aim of this retrospective study was to assess pre-analytical variables that might influence the diagnostic utility of FC of lymph node (LN) fine needle aspirate (FNA) specimens from dogs with lymphoproliferative diseases. The study included 987 cases with LN FNA specimens sent for immunophenotyping that were submitted to a diagnostic laboratory in Italy from 2009 to 2015. Cases were grouped into 'diagnostic' and 'non-diagnostic'...
February 2018: Veterinary Journal
Praveen Vimalathas, Alexandra Farris, Dorothea Letner, Vikram Deshpande, Vijay Yajnik, Wayne Shreffler, John Garber
Eosinophilic esophagitis (EoE) is increasingly recognized allergic disease associated with dysphagia and esophageal fibrosis. We aimed to determine expression patterns of specific eosinophil integrins that promote eosinophilic infiltration of the esophageal epithelium, and to determine how key EoE-related cytokines influence eosinophil activation and survival. Esophageal and peripheral eosinophils were isolated from 20 adult subjects with EoE for immunophenotyping and integrin profiling using multicolor flow cytometry and immunohistochemistry...
February 9, 2018: Immunology and Cell Biology
Giuseppe Cappellano, Christian Ploner, Susanne Lobenwein, Sieghart Sopper, Paul Hoertnagl, Christina Mayerl, Nikolaus Wick, Gerhard Pierer, Georg Wick, Dolores Wolfram
The most common complication of silicone breast implants is capsular contracture (massive scar formation around the implant). We postulate that capsular contracture is always a sequel to inflammatory processes, with both innate and adaptive immune mechanisms participating. In general, fibroblasts and macrophages have been used as cell types to evaluate in vitro the biocompatibility of breast implant surfaces. Moreover, also T cells have been found at the implant site at the initial stage of fibrous capsule formation...
2018: PloS One
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