Main description:

This volume serves as a reference for the dissemination of advances made in the study of Hepatitis B Virus (HBV). Hepatitis B Virus: Methods and Protocols details protocols and techniques ranging from cell culture studies to in vivo and clinical immunology. The chapters in this book discuss treatments of in vitro infection systems, analysis and quantification of cccDNA and its mutations; in vitro polymerase activity assays; cellular trafficking of core proteins; intracellular calcium metabolism; detection, cloning, and sequencing of HBV markers; and new strategies aimed at exploiting new mechanisms for drug discovery. The book also covers classical methods for resolution of extracellular viral particles by native gel electrophoresis, and methods for detecting HBV antigens in drug discovery. 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, Hepatitis B Virus: Methods and Protocols is a valuable tool for researchers to use toward their advanced studies in HBV.

Contents:

Preface...
Table of Contents...
Contributing Authors...

1. NTCP-Reconstituted In Vitro HBV Infection System
Yinyan Sun, Yonghe Qi, Bo Peng, and Wenhui Li

2. Hepatitis B Virus Infection of HepaRG Cells, HepaRG-hNTCP Cells and Primary Human Hepatocytes
Yi Ni and Stephan Urban

3. Live Cell Imaging Confocal Microscopy Analysis of HBV Myr-PreS1 Peptide Binding and Uptake in NTCP-GFP Expressing HepG2 Cells
Alexander Koenig and Dieter Glebe

4. Intracytoplasmic Transport of Hepatitis B Virus Capsids
Quentin Osseman and Michael Kann5. A Homokaryon Assay for Nucleocytoplasmic Shuttling Activity of HBV Core Protein
Ching-Chun Yang, Hung-Cheng Li, and Chiaho Shih

6. Analyses of HBV cccDNA Quantification and Modification
Yuchen Xia, Daniela Stadler, Chunkyu Ko, and Ulrike Protzer

7. Detection of HBV cccDNA Methylation from Clinical Samples by Bisulfite Sequencing and Methylation Specific PCR
Yongmei Zhang, Richeng Mao, Haitao Guo, and Jiming Zhang

8. A T7 Endonuclease I Assay to Detect TALEN-Mediated Mutation of HBV cccDNA
Kristie Bloom, Abdullah Ely, and Patrick Arbuthnot9. Detection of Hepatocyte Clones Containing Integrated Hepatitis B Virus DNA using Inverse Nested PCR
Thomas Tu and Allison R. Jilbert

10. Highly Sensitive Detection of HBV RNA in Liver Tissue by In Situ Hybridization
Diego Calabrese and Stefan F. Wieland

11. Immunofluorescent Staining for the Detection of the Hepatitis B Core Antigen in Frozen Liver Sections of Human Liver Chimeric Mice
Lena Allweiss, Marc Lutgehetmann, and Maura Dandri

12. Measuring Changes in Cytosolic Calcium Levels in HBV- and HBx-Expressing Cultured Primary Hepatocytes
Jessica C. Casciano and Michael J. Bouchard

13. In Vitro Assays for RNA Binding and Protein Priming of Hepatitis B Virus Polymerase
Daniel N. Clark, Scott A. Jones, and Jianming Hu

14. In Vitro Enzymatic and Cell Culture-Based Assays for Measuring Activity of HBV RNaseH Inhibitors
Elena Lomonosova and John E. Tavis<

15. Detection of Hepatitis B Virus Particles Released from Cultured Cells by Particle Gel Assay
Ran Yan, Dawei Cai, Yuanjie Liu, and Haitao Guo

16. Microtiter-Format Assays for HBV Antigen Quantitation in Non-Clinical Applications
Cally Goddard, Lale-Bildrici Ertekin, Xiahoe Wang, and Andrea Cuconati17. Deep Sequencing of the Hepatitis B Virus Genome: Analysis of Multiple Samples by Implementation of the Illumina Platform
Quan-Xin Long, Jie-Li Hu, and Ai-Long Huang

18. Generation of Replication Competent Hepatitis B Virus Genome from Blood Samples for Functional Characterization
Yanli Qin, Yong-Xiang Wang, Jiming Zhang, Jisu Li, and Shuping Tong

19. Hydrodynamic HBV Transfection Mouse Model
Li-Ling Wu, Hurng-Yi Wang, and Pei-Jer Chen

20. An ELISPOT-Based Assay to Measure HBV-Specific CD8+ T Cell Responses in Immunocompetent Mice
Tracy D. Reynolds, Safiehkhatoon Moshkani, and Michael D. Robek

21. Advanced Method for Isolation of Mouse Hepatocytes, Liver Sinusoidal Endothelial Cells and Kupffer Cells
Jia Liu, Xuan Huang, Melanie Werner, Ruth Broering, Dongliang Yang, and Mengji Lu

22. Partial Hepatectomy and Castration of HBV Transgenic Mice
Yongjun Tian and Jing-hsiung James Ou

23. Studying HBV Infection and Therapy in Immune Deficient NOD-Rag1-/-IL2RgammaC-null (NRG) Fumarylacetoacetate Hydrolase (Fah) Knockout Mice Transplanted with Human Hepatocytes
Feng Li, Kouki Nio, Fumihiko Yasui, Christopher M. Murphy, and Lishan Su

24. Measurement of Antiviral Effect and Innate Immune Response during Treatment of Primary Woodchuck Hepatocytes
Marta G. Murreddu, Manasa Suresh, Severin O. Gudima, and Stephan Menne