Methods in Molecular Biology 1591 George Edward Rainger Helen M. Mcgettrick Editors T-Cell Traffi cking Methods and Protocols Second Edition M M B ethods in olecular iology Series Editor John M. Walker School of Life and Medical Sciences University of Hertfordshire Hatfield, Hertfordshire, AL10 9AB, UK For further volumes: http://www.springer.com/series/7651 T-Cell Trafficking Methods and Protocols Second Edition Edited by George Edward Rainger Institute of Cardiovascular Sciences, University of Birmingham, Birmingham, UK Helen M. Mcgettrick Institute of Inflammation and Ageing, College of Medicine and Dental Sciences, University of Birmingham, Birmingham, UK Editors George Edward Rainger Helen M. Mcgettrick Institute of Cardiovascular Sciences Institute of Inflammation and Ageing University of Birmingham College of Medicine and Dental Sciences Birmingham, UK University of Birmingham Birmingham, UK ISSN 1064-3745 ISSN 1940-6029 (electronic) Methods in Molecular Biology ISBN 978-1-4939-6929-6 ISBN 978-1-4939-6931-9 (eBook) DOI 10.1007/978-1-4939-6931-9 Library of Congress Control Number: 2017932550 © Springer Science+Business Media LLC 2017 This work is subject to copyright. All rights are reserved by the Publisher, whether the whole or part of the material is concerned, specifically the rights of translation, reprinting, reuse of illustrations, recitation, broadcasting, reproduction on microfilms or in any other physical way, and transmission or information storage and retrieval, electronic adaptation, computer software, or by similar or dissimilar methodology now known or hereafter developed. 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Preface Welcome to the second edition of Methods and Protocols for assessing T cell Trafficking, in the Methods in Molecular Biology series. The trafficking of T cells is relevant in numer- ous contexts. It occurs during population and maturation of T cells in the thymus and is required for dissemination of antigen naïve T cells to the secondary lymphatic organs where immune responses are initiated. Indeed, T cell trafficking within lymph nodes plays an important role in the maturation of primary and secondary immune responses. Antigen experienced effector T cells can undertake compartmentalized recirculation during immune surveillance. In addition they are recruited to tissues during inflammation where they play important roles in the inflammatory response. Importantly, we now recognize that inap- propriate or persistent trafficking of T cells into such sites makes a major contribution to the pathogenesis of immune-mediated inflammatory diseases which have an autoimmune or chronic inflammatory component. Thus the trafficking of T cells has both physiological and pathological relevance and provides some challenging environments in which to make quantitative measurements. This has seen the development of expertise which goes well beyond the standard laboratory methodologies which can be supported by commercially available kits and reagents. The methods in this edition have been developed by experts in T cell trafficking, who have spent many years perfecting them. Each chapter contains a step-by-step guide to conducting the assays, with useful hints to avoid common pitfalls. The volume is organized into three sec- tions. The first addresses homeostatic T cell trafficking during thymic maturation, followed by the subsequent colonization of and egress from secondary lymphoid organs. The second addresses T cell trafficking during “normal” inflammatory and immune responses. Lastly, we include a section on T cell trafficking in disease. Each section is headed by an informa- tive and accessible introduction written by experts who are actively investigating the regula- tion of T cell trafficking in these different scenarios. We believe this will ensure that this book will become an essential point of reference for those new to the field of T cell traffick- ing, or to those looking to expand their technical capabilities. We would like to thank all the authors for their invaluable contributions and willingness to share their expertise. Thanks also to Professor John Walker, the series editor, for guid- ance in the process of compiling the book. G. Ed. Rainger is generously supported by the British Heart Foundation of the UK. Helen M. Mcgettrick is supported by generous funds from Arthritis Research UK. Birmingham, UK George Edward Rainger Helen M. Mcgettrick v Contents Preface.......................................................... v Contributors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ix 1 Introduction to Homeostatic Migration ............................. 1 Mark C. Coles 2 Analysis of Thymocyte Migration, Cellular Interactions, and Activation by Multiphoton Fluorescence Microscopy of Live Thymic Slices ........... 9 Jessica N. Lancaster and Lauren I.R. Ehrlich 3 Visualizing and Tracking T Cell Motility In Vivo....................... 27 Robert A. Benson, James M. Brewer, and Paul Garside 4 Graph Theory-Based Analysis of the Lymph Node Fibroblastic Reticular Cell Network ................................................. 43 Mario Novkovic, Lucas Onder, Gennady Bocharov, and Burkhard Ludewig 5 Visualizing Endogenous Effector T Cell Egress from the Lymph Nodes ..... 59 Manisha Menon, Alexandre P. Benechet, and Kamal M. Khanna 6 Introduction: T Cell Trafficking in Inflammation and Immunity........... 73 Myriam Chimen, Bonita H.R. Apta, and Helen M. Mcgettrick 7 Leukocyte Adhesion Under Hemodynamic Flow Conditions.............. 85 Charlotte Lawson, Marlene Rose, and Sabine Wolf 8 Endocrine Regulation of Lymphocyte Trafficking In Vitro ............... 101 Bonita H.R. Apta, Myriam Chimen, and Helen M. Mcgettrick 9 Mesenchymal Stromal Cells as Active Regulators of Lymphocyte Recruitment to Blood Vascular Endothelial Cells....................... 121 Helen M. Mcgettrick, Lewis S.C. Ward, George Edward Rainger, and Gerard B. Nash 10 Monitoring RhoGTPase Activity in Leukocytes Using Classic “Pull-Down” Assays ............................................ 143 Marouan Zarrouk, David Killock, Izajur Rahman, Jessica Davies, and Aleksandar Ivetić 11 Utilizing Lentiviral Gene Transfer in Primary Endothelial Cells to Assess Lymphocyte-Endothelial Interactions........................ 155 Jasmeet S. Reyat, Michael G. Tomlinson, and Peter J. Noy 12 Introduction to Lymphocyte Trafficking in Disease..................... 169 Patricia F. Lalor and Elizabeth A. Hepburn 13 Using Ex Vivo Liver Organ Cultures to Measure Lymphocyte Trafficking.... 177 Benjamin G. Wiggins, Zania Stamataki, and Patricia F. Lalor vii viii Contents 14 In Vitro and Ex Vivo Models to Study T Cell Migration Through the Human Liver Parenchyma..................................... 195 Benjamin G. Wiggins, Konstantinos Aliazis, Scott P. Davies, Gideon Hirschfield, Patricia F. Lalor, Gary Reynolds, and Zania Stamataki 15 Monitoring Migration of Activated T Cells to Antigen-Rich Non-lymphoid Tissue . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 215 Eleanor Jayne Ward, Hongmei Fu, and Federica Marelli-Berg 16 Tissue Digestion for Stromal Cell and Leukocyte Isolation ............... 225 Saba Nayar, Joana Campos, Nathalie Steinthal, and Francesca Barone 17 T Cell Response in the Lung Following Influenza Virus Infection.......... 235 Robert A. Benson, Jennifer C. Lawton, and Megan K.L. MacLeod Index . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 249 Contributors Konstantinos aliazis • Centre for Liver Research, Institute for Immunology and Immunotherapy, University of Birmingham, Birmingham, UK Bonita H.R. apta • Institute of Cardiovascular Sciences, College of Medicine and Dental Sciences, University of Birmingham, Birmingham, UK FRancesca BaRone • Centre for Translational Inflammation Research, Institute of Inflammation and Ageing, College of Medical & Dental Sciences, University of Birmingham Research Laboratories, Queen Elizabeth Hospital, Birmingham, UK alexandRe p. BenecHet • Division of Immunology, Transplantation and Infectious Diseases, IRCCS San Raffaele Scientific Institute, Milan, Italy RoBeRt a. Benson • Centre for Immunobiology, Institute of Infection, Immunity and Inflammation, The University of Glasgow, Glasgow, UK FedeRica MaRelli-BeRg • William Harvey Research Institute—Heart Centre, Barts and the London School of Medicine and Dentistry, Queen Mary University of London, London, UK gennady BocHaRov • Institute of Numerical Mathematics, Russian Academy of Sciences, Moscow, Russian Federation JaMes M. BReweR • Institute of Infection, Immunity and Inflammation, College of Medical, Veterinary and Life Sciences, Glasgow Biomedical Research Centre, University of Glasgow, Glasgow, UK Joana caMpos • Centre for Translational Inflammation Research, Institute of Inflammation and Ageing, College of Medical & Dental Sciences, University of Birmingham Research Laboratories, Queen Elizabeth Hospital, Birmingham, UK MyRiaM cHiMen • Institute of Cardiovascular Sciences, College of Medicine and Dental Sciences, University of Birmingham, Birmingham, UK MaRK c. coles • Department of Biology, Centre for Immunology and Infection, University of York, North Yorkshire, UK scott p. davies • Centre for Liver Research, Institute for Immunology and Immunotherapy, University of Birmingham, Birmingham, UK Jessica davies • Cytoskeleton/Membrane Signalling Research Group, Cardiovascular Division, King’s College London, London, UK lauRen i.R. eHRlicH • Department of Molecular Biosciences, Institute for Cellular and Molecular Biology, The University of Texas at Austin, Austin, TX, USA HongMei Fu • William Harvey Research Institute—Heart Centre, Barts and the London School of Medicine and Dentistry, Queen Mary University of London, London, UK paul gaRside • Institute of Infection, Immunity and Inflammation, College of Medical, Veterinary and Life Sciences, Glasgow Biomedical Research Centre, Wellcome Trust Centre for Molecular Parasitology, Glasgow, UK elizaBetH a. HepBuRn • Department of Cellular Pathology, Cheltenham General Hospital, Cheltenham, UK gideon HiRscHField • Centre for Liver Research, Institute for Immunology and Immunotherapy, University of Birmingham, Birmingham, UK ix x Contributors aleKsandaR ivetić • Cytoskeleton/Membrane Signalling Research Group, Cardiovascular Division, King’s College London, London, UK KaMal M. KHanna • Department of Immunology, University of Connecticut Health, Farmington, CT, USA david KillocK • Cytoskeleton/Membrane Signalling Research Group, Cardiovascular Division, King’s College London, London, UK patRicia F. laloR • Centre for Liver Research, Immunity and Immunotherapy, Institute of Biomedical Research, University of Birmingham, Birmingham, UK Jessica n. lancasteR • Department of Molecular Biosciences, Institute for Cellular and Molecular Biology, The University of Texas at Austin, Austin, TX, USA cHaRlotte lawson • Comparative Biomedical Sciences, Royal Veterinary College, London, UK JenniFeR c. lawton • Centre for Immunobiology, Institute of Infection, Immunity and Inflammation, The University of Glasgow, Glasgow, UK BuRKHaRd ludewig • Institute of Immunobiology, Kantonsspital St . Gallen, St . Gallen, Switzerland Megan K.l. MacLeod • Centre for Immunobiology, Institute of Infection, Immunity and Inflammation, The University of Glasgow, Glasgow, UK Helen M. McgettRicK • Institute of Inflammation and Ageing, College of Medicine and Dental Sciences, University of Birmingham, Birmingham, UK ManisHa Menon • Department of Immunology , University of Connecticut Health, Farmington, CT, USA geRaRd B. nasH • Institute of Cardiovascular Sciences, University of Birmingham, Birmingham, UK saBa nayaR • Centre for Translational Inflammation Research, Institute of Inflammation and Ageing, College of Medical & Dental Sciences, University of Birmingham Research Laboratories, Queen Elizabeth Hospital, Birmingham, UK MaRio novKovic • Institute of Immunobiology, Kantonsspital St . Gallen, St . Gallen, Switzerland peteR J. noy • School of Biosciences, College of Life and Environmental Sciences, University of Birmingham, Birmingham, UK lucas ondeR • Institute of Immunobiology, Kantonsspital St . Gallen, St . Gallen, Switzerland izaJuR RaHMan • Cytoskeleton/Membrane Signalling Research Group, Cardiovascular Division, King’s College London, London, UK geoRge edwaRd RaingeR • Institute of Cardiovascular Sciences, University of Birmingham, Birmingham, UK JasMeet s. Reyat • School of Biosciences, College of Life and Environmental Sciences, University of Birmingham, Birmingham, UK gaRy Reynolds • Centre for Liver Research, Institute for Immunology and Immunotherapy, University of Birmingham, Birmingham, UK MaRlene Rose • Harefield Hospital, Imperial College, London, UK zania staMataKi • Centre for Liver Research, Immunity and Immunotherapy, Institute of Biomedical Research, University of Birmingham, Birmingham, UK natHalie steintHal • Centre for Translational Inflammation Research, Institute of Inflammation and Ageing, College of Medical & Dental Sciences, University of Birmingham Research Laboratories, Queen Elizabeth Hospital, Birmingham, UK Contributors xi MicHael g. toMlinson • School of Biosciences, College of Life and Environmental Sciences, University of Birmingham, Birmingham, UK eleanoR Jayne waRd • William Harvey Research Institute—Heart Centre, Barts and the London School of Medicine and Dentistry, Queen Mary University of London, London, UK lewis s.c. waRd • Institute of Inflammation and Ageing, University of Birmingham, Birmingham, UK BenJaMin g. wiggins • Centre for Liver Research, Immunity and Immunotherapy, Institute of Biomedical Research, University of Birmingham, Birmingham, UK saBine wolF • Comparative Biomedical Sciences, Royal Veterinary College, London, UK MaRouan zaRRouK • Cytoskeleton/Membrane Signalling Research Group, Cardiovascular Division, King’s College London, London, UK