Full Description
This volume explores data from the applications of molecular biological methods and the applications of recent immunological and cytogenetic methods in Epstein-Barr Virus (EBV) that will offer readers possible new solutions to the unresolved problems in the EBV field. Chapters in this book cover topics such as: viral life cycle, latency, EBV-associated diseases and EBV diagnostics; in vitro methods including organotypic cultures for the analysis of EBV-epithelial cell interactions; identification of the interacting viral and cellular proteins using affinity purification-mass spectrometry methods; 3D telomere FISH; transcription analysis using high-throughput RNA sequencing, qPCR and nuclear run-on assay; analysis of viral and cellular microRNAs; isolation and characterization of exosomes and the assessment of their function; characterization of the viral genome by terminal repeat analysis and sequencing; the use of chromatin immunoprecipitation coupled sequencing (ChIP-Seq) for theanalysis of Zta-DNA interactions; epigenetic analysis by bisulfite sequencing and ChIP; novel in vivo models for the study of EBV infection; and how immunological, virological, tissue culture and molecular methods can be combined to yield Good Manufacturing Practice-compliant EBV-specific T cells for the immunotherapy of EBV-associated post-transplant lymphoproliferative diseases (PTLD). Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Cutting-edge and comprehensive, Epstein Barr-Virus: Methods and Protocols is a valuable resource for anyone who is interested in this fascinating and evolving field.
Contents
Current Trends and Alternative Scenarios in EBV Research.- Epstein-Barr Virus: Clinical Diagnostics.- Establishment of EBV-Infected Lymphoblastoid Cell Lines.- Generation and Infection of Organotypic Cultures with Epstein-Barr Virus.- Affinity Purification-Mass Spectroscopy Methods for Identifying Epstein-Barr Virus-Host Interactions.- The Use of 3D Telomere FISH for the Characterization of the Nuclear Architecture in EBV-Positive Hodgkin's Lymphoma.- Analysis of EBV Transcription Using High-Throughput RNA Sequencing.- Analysis of Viral Promoter Usage in EBV-Infected Cell Lines: A Comparison of qPCR following Conventional RNA Isolation and Nuclear Run-On Assay.- Analysis of Viral and Cellular MicroRNAs in EBV Infected Cells.- Isolation and Characterization of Exosomes Released by EBV Immortalized Cells.- Functional Analysis of Exosomes Derived from EBV-Infected Cells.- Terminal Repeat Analysis of EBV Genomes.- Characterization of EBVPromoters and Coding Regions by Sequencing PCR-Amplified DNA Fragments.- The Use of Chromatin Precipitation Coupled to DNA Sequencing (ChIP-Seq) for the Analysis of Zta Binding to the Human and EBV Genome.- Analysis of Viral Epigenotypes Using Bisulfite Sequencing: A Detailed Protocol for the Crucial Bisulfite Modification and PCR Amplification Steps.- Analysis of Viral Epigenotypes using Chromatin Immunoprecipitation.- Mice with Reconstituted Human Immune System Components as a Tool to Study Immune Cell Interactions in EBV Infection.- Generation and Analysis of Humanized Mouse Model of EBV Infection.- EBV-Directed T Cell Therapeutics for EBV-Associated Lymphomas.
