Description
Hands-on laboratory experts describe for the novice investigator a host of novel technologies and molecular techniques specifically designed to study cellular immunology and promote its gene therapy applications. Presented in step-by-step detail to ensure ready reproducibility, these protocols range from flow cytometric techniques to detect cytokines and growth factors in different specimens, to methods for generating and expanding dendritic cells and hematopoietic progenitors from different origins. Although the protocols require little previous experience, even highly skilled researchers will augment their skills with many novel timesaving techniques. Part I. Introduction Primary Immunodeficiencies Caused by Defects of Cytokines and Cytokine Receptors Volker Wahn Part II. Detection Assays for Cytokines and Growth Factors Intracellular Detection of T-Cell Cytokines: Differentiation of TH1 and TH2 Cells Ursula Banning and Dieter Krholz Analysis of Cytokine Profiles in Human Skin Jean Krutmann, Renz Mang, and Susanne Grether-Beck Intracytoplasmic Detection of Proinflammatory Cytokines and Chemokines in Monocytes by Flow Cytometry Christian Schultz Flow-Cytometric Immune Function Methodology for Human Peripheral Blood Dendritic Cells Kerstin Willmann Evaluation of the Frequency of Virus-Specific CD8+ T Cells by Cytokine Flow Cytometry Bodo Hoffmeister, Felix Kiecker, Ingolf Surel, Elham Khatamzas, Volker Schuster, Hans-Dieter Volk, and Florian Kern Detection of Common Cytokine and Colony Stimulating Factor Gene Polymorphisms Thomas Lehrnbecher and Stephen J. Chanock Apoptosis Induction by TRAIL Angelika Eggert, Hauke Sieverts, and Henning Walczak Angiogenic Cytokines: Quantitative and Functional Analysis Angelika Eggert and John H. Maris Part III. Protocols on Gene Therapy Experimental Strategies for Combined Suicide and Immune Cancer Gene Therapy: An Overview Christof M. Kramm, Tim Niehues, and Nikolai G. Rainov Gene Therapy with Plasmids Encoding Cytokine- or Cytokine Receptor-IgG Chimeric Proteins Ciriaco A. Piccirillo and Grald J. Prud’homme Interferon-b Gene Therapy for the Treatment of Arthritis Yuti Chernajovsky, Hanna Dreja, Kostas Triantaphyllopoulos, and David Gould Gene Gun-Based In Vivo Gene Transfer: Application to DNA Vaccination Tohru Sakai and Kunisuke Himeno Purification of the Eukaryotic Heat-Shock Proteins Hsp70 and gp96 Arne von Bonin, Solveig H. Mor, and Minka Breloer Genetic Engineering of a Recombinant Fusion Protein Possessing an Antitumor Antibody Fragment and a TNF-a Moiety Jim Xiang and John R. Gordon Genetic Engineering of Dendritic Cells by Adenovirus-Mediated TNF-a Gene Transfer Jim Xiang and Josh Wu Retroviral Transfer of T-Cell Receptor Genes into Human Peripheral Blood Lymphocytes Timothy M. Clay and Michael I. Nishimura Gene Transfer to Articular Chondrocytes with Recombinant Adenovirus Glyn D. Palmer, Elvire Gouze, Jean-Noel Gouze, Oliver B. Betz, Christopher H. Evans, and Steven C. Ghivizzani Gene Therapy of X-Linked Severe Combined Immunodeficiency Salima Hacein-Bey-Abina, Genevieve de Saint Basile, and Marina Cavazzana-Calvo Gene Transfer for Generation of Tumor and Leukemia Vaccines Dagmar Dilloo and Andree Zibert Cytokine Gene Delivery into the Central Nervous System Using Intrathecally Injected Nonreplicative Viral Vectors Roberto Furlan, Stefano Pluchino, Peggy C. Marconi, and Gianvito Martino Part IV. Ex Vivo Cell Expansion In Vitro Regulation of Colony Stimulating Factor-Mediated Hematopoiesis in Healthy Individuals and Patients with Different Types of Myeloproliferative Disease Thomas Vraetz, Peter D. Emanuel, and Charlotte M. Niemeyer In Vitro Generation of Dendritic Cells from Cord Blood CD34+ Hematopoietic Progenitors Cells Bruno Canque, Michele Rosenzwajg, and Jean Claude Gluckman Generation of Transgenic T Cells from Human CD34+ Cord Blood Cells Bruno Verhasselt, Evelien Naessens, and Veronique Stove Ex Vivo Expansion of Umbilical Cord Blood Cells on Feeder Layers Martin Bornhuser Ex Vivo Production of Cord Blood CD34+ Derived Myeloid Precursors After Serum-Free Static Culture Sergio Querol Ex Vivo Expansion of Hematopoietic Stem Cells Beatriz Albella, Jos Carlos Segovia, Guillermo Guenechea, and Juan Antonio Bueren Large-Scale Ex Vivo Expansion of Human Megakaryocytes for Clinical Use Phil Lefebvre and Isaac Cohen Cytokine-Mediated Expansion of Human NOD-SCID-Repopulating Cells Kohichiro Tsuji, Takahiro Ueda, and Yasuhiro Ebihara Part V. Maturation and Differentiation of Dendritic Cells Differentiation of Human Antigen-Presenting Dendritic Cells from CD34+ Hematopoietic Stem Cells In Vitro Xin-Sheng Ju and Martin Zenke Dendritic Cell Development from Mobilized Peripheral Blood CD34+ Cells Deanne M. R. Lathers, Nicholas Achille, and M. Rita I. Young In Vitro Maturation of Dendritic Cells from Blood Progenitors Lina Matera and Alessandra Galetto. Generation of Human Type 1- and Type 2-Polarized Dendritic Cells from Peripheral Blood Pawel Kalinski, Pedro Vieira, Joost H. N. Schuitemaker, Quan Cai, and Martien Kapsenberg Preparation of Human Dendritic Cells for Tumor Vaccination Michael R. Shurin Generation of Leukemic Dendritic Cells from Patients with Acute Myeloid Leukemia Mohamad Mohty, Batrice Gaugler, and Daniel Olive Index
