CLINICAL FLOW CYTOMETRY – EMERGING APPLICATIONS Edited by Ingrid Schmid Clinical Flow Cytometry – Emerging Applications Edited by Ingrid Schmid Published by InTech Janeza Trdine 9, 51000 Rijeka, Croatia Copyright © 2012 InTech All chapters are Open Access distributed under the Creative Commons Attribution 3.0 license, which allows users to download, copy and build upon published articles even for commercial purposes, as long as the author and publisher are properly credited, which ensures maximum dissemination and a wider impact of our publications. After this work has been published by InTech, authors have the right to republish it, in whole or part, in any publication of which they are the author, and to make other personal use of the work. Any republication, referencing or personal use of the work must explicitly identify the original source. As for readers, this license allows users to download, copy and build upon published chapters even for commercial purposes, as long as the author and publisher are properly credited, which ensures maximum dissemination and a wider impact of our publications. Notice Statements and opinions expressed in the chapters are these of the individual contributors and not necessarily those of the editors or publisher. No responsibility is accepted for the accuracy of information contained in the published chapters. The publisher assumes no responsibility for any damage or injury to persons or property arising out of the use of any materials, instructions, methods or ideas contained in the book. Publishing Process Manager Martina Blecic Technical Editor Teodora Smiljanic Cover Designer InTech Design Team First published April, 2012 Printed in Croatia A free online edition of this book is available at www.intechopen.com Additional hard copies can be obtained from orders@intechopen.com Clinical Flow Cytometry – Emerging Applications, Edited by Ingrid Schmid p. cm. ISBN 978-953-51-0575-6 Contents Preface IX Chapter 1 Effect of Monocyte Locomotion Inhibitory Factor (MLIF) on the Activation and Production of Intracellular Cytokine and Chemokine Receptors in Human T CD4 + Lymphocytes Measured by Flow Cytometry 1 Sara Rojas-Dotor Chapter 2 Applications of Flow Cytometry to Clinical Microbiology 17 Barbara Pieretti, Annamaria Masucci and Marco Moretti Chapter 3 High-Throughput Flow Cytometry for Predicting Drug-Induced Hepatotoxicity 43 Marion Zanese, Laura Suter, Adrian Roth, Francesca De Giorgi and François Ichas Chapter 4 B Cells in Health and Disease – Leveraging Flow Cytometry to Evaluate Disease Phenotype and the Impact of Treatment with Immunomodulatory Therapeutics 60 Cherie L. Green, John Ferbas and Barbara A. Sullivan Chapter 5 Evaluation of the Anti-Tumoural and Immune Modulatory Activity of Natural Products by Flow Cytometry 91 Susana Fiorentino, Claudia Urueña, Sandra Quijano, Sandra Paola Santander, John Fredy Hernandez and Claudia Cifuentes Chapter 6 Identification and Characterization of Cancer Stem Cells Using Flow Cytometry 107 Yasunari Kanda Chapter 7 Flow Based Enumeration of Plasmablasts in Peripheral Blood After Vaccination as a Novel Diagnostic Marker for Assessing Antibody Responses in Patients with Hypogammaglobulinaemia 125 Vojtech Thon, Marcela Vlkova, Zita Chovancova, Jiri Litzman and Jindrich Lokaj VI Contents Chapter 8 Applications of Flow Cytometry in Solid Organ Allogeneic Transplantation 143 Dimitrios Kirmizis, Dimitrios Chatzidimitriou, Fani Chatzopoulou, Lemonia Skoura and Gregory Myserlis Chapter 9 The Use of Flow Cytometry to Monitor T Cell Responses in Experimental Models of Graft-Versus-Host Disease 151 Bryan A. Anthony and Gregg A. Hadley Chapter 10 Lymphocyte Apoptosis, Proliferation and Cytokine Synthesis Pattern in Children with Helicobacter pylori Infection 173 Anna Helmin-Basa, Lidia Gackowska, Izabela Kubiszewska, Malgorzata Wyszomirska-Golda, Andrzej Eljaszewicz, Grazyna Mierzwa, Anna Szaflarska-Poplawska, Mieczyslawa Czerwionka-Szaflarska, Andrzej Marszalek and Jacek Michalkiewicz Chapter 11 The Effect of Epigallocatechin Gallate (EGCG) and Metal Ions Corroded from Dental Casting Alloys on Cell Cycle Progression and Apoptosis in Cells from Oral Tissues 191 Jiansheng Su, Zhizen Quan, Wenfei Han, Lili Chen and Jiamei Gu Preface Advances in patient management have often been closely linked to the development of critical quantitative analysis methods. Flow cytometry is such an important methodology. It can be applied to individual cells or organelles allowing investigators interested in obtaining information about the functional properties of cells to assess the differences among cells in a heterogeneous cell preparation or between cells from separate samples. It is characterized by the use of a select wavelength of light (or multiple ones) to interrogate cells or other particles one at a time providing statistically relevant, rapid correlated measurements of multiple parameters with excellent temporal resolution. These intrinsic attributes, as well as advances in instrumentation and fluorescent probes and reagents, have contributed to the tremendous growth of clinical applications of flow cytometry and to the world-wide expansion of laboratories which use this technology since its inception in the late 1960s. This publication reflects these facts as indicated by the global author panel and the wide range of sample types, assays, and methodologies described. Openly accessible, the book is intended to introduce novices to this powerful technology and also provide experienced professionals with valuable insights and an opportunity to refresh or up- date their knowledge in various subject areas of clinical flow cytometry. Ingrid Schmid, Mag. Pharm. Department of Medicine Division of Hematology-Oncology University of California, Los Angeles USA [...]... studies d Applications 347-436 (49 0–5 50)-(54 0–6 30) 508-(52 5–5 90) Depends on fluorochrome conjugated 340-(39 8–4 85) 340-549 469-545 (46 0–5 10 )–( 52 0–6 10) 510-(58 7–6 35) 484-501 488-525 507-529 Depends on fluorochrome conjugated Excitation/ Emission Wavelength (λmax)nm 504-525 536-625 510-595 488-509 340-450 425-550 497-563 495-525 580-620 496-526 20 Clinical Flow Cytometry – Emerging Applications Applications... complemented by technologies such as those provided by flow cytometry We have gotten used to consider the flow cytometry applicable only in the field of hematology, then associate it with clinical microbiology makes it even more mysterious Over the past forty years we have witnessed several attempts of application of the flow cytometry to microbiology, with good results but also with many difficulties 18 Clinical. .. provides information regarding the cell’s state of activation Chemokine receptors can by analyzed by flow cytometry using fluorescently labeled anti-receptor antibodies or fluorescently-labeled chemokines Combining these reagents with antibodies against the activation marker CD69 8 Clinical Flow Cytometry – Emerging Applications enables analysis of cell activation within specific cell population Figure... results but also with many difficulties 18 Clinical Flow Cytometry – Emerging Applications In particular, the problems encountered relate the difficulty of measuring microbes by flow due to their small size and point towards the development of instrumentation that has managed to overcome this limitation of standard instrumentation used for routine flow cytometry in different fields from microbiology The... (Utrera-Barillas et al., 2003) The p65–p50 heterodimer comprises the most abundant form of NF-κB in a PMA-induced system Temporary studies showed that MLIF induces p50 translocation, which may be explained 2 Clinical Flow Cytometry – Emerging Applications by the ability of MLIF to induce AMPc synthesis and protein kinase A phosphorylation in NF-κB and IκB followed by NF-κB translocation (Kretschmer et... measuring changes in cells surface 6 Clinical Flow Cytometry – Emerging Applications markers and assays for measuring the expression of intracellular cytokine Flow cytometers are laser-based cell counters that are capable of distinguishing 3, 4, 5 or more (depending of flow cytometer), different fluorescence emissions, each associated with a particle identified by its light scatter proprieties Fluorescence... microflow cytometer can detect for Escherichia coli, Listeria, and Salmonella 103, 105, and 104 cfu/mL, respectively, while the limits of detection for the toxins as cholera toxin, staphylococcal enterotoxin B, and ricin were 1.6, 0.064, and 1.6 ng/mL respectively (Kim, 2009) 26 Clinical Flow Cytometry – Emerging Applications 3.1.2 Fungi The use of FCM to detect fungal pathogens was first described by. .. are often found in damaged tissue (Luscinskas & Gimbrone, 1996) 4 Clinical Flow Cytometry – Emerging Applications Chemokines are small polypeptides that activate and direct the migration of monocytes, neutrophils, eosinophils and activated T lymphocytes from the bloodstream to sites of infection They also regulate pro-inflammatory signals by binding to specific receptors belonging to the superfamily of... directed against IL-1β, IL-2, IFN-γ, IL-4, and IL-10, and Th1 and Th2-associated cytokine-producing lymphocytes can be counted on a flow cytometer This procedure helps to differentiate between Th1 (IFN-γ producing) and Th2 (IL-4 producing) 10 Clinical Flow Cytometry – Emerging Applications cells in specific cell populations MLIF increased the expression of IL-lβ, IL-2, IFN-γ, IL-4, and IL-10 Following PMA+MLIF... light, and the results represent cumulative individual cytometric characteristics An important analytical feature of flow cytometers is their ability to measure multiple cellular parameters (analytical flow Applications of Flow Cytometry to Clinical Microbiology 19 cytometers) Some flow cytometers are able to physically separate cell subsets (sorting) based on their cytometric characteristics (cell . CLINICAL FLOW CYTOMETRY – EMERGING APPLICATIONS Edited by Ingrid Schmid Clinical Flow Cytometry – Emerging Applications Edited. be obtained from orders@intechopen.com Clinical Flow Cytometry – Emerging Applications, Edited by Ingrid Schmid p. cm. ISBN 978-953-51-0575-6