HIV Protocols

ISBN-13: 9781493930456 / Angielski / Twarda / 2015 / 383 str.

Vinayaka R. Prasad; Ganjam Kalpana

Opis

This third edition volume expands upon the previous editions with new research topics and numerous leading-edge HIV protocols. This book contains chapters that cover various aspects of HIV-1 replication and are helpful for investigating the mechanisms of HIV-1 replication or pathogenesis. The chapters are grouped into six sections: HIV early events; HIV-1 RNA structure and RNA-protein interactions; HIV-1 post-integration events; HIV-1 pathogenesis in animal models; tools to study HIV-1 latency; and Neuro AIDS. Chapters are written by research experts who have diligently developed techniques, improved them, simplified them, or applied them to their own HIV research. Written in the highly successful Methods in Molecular Biology series format, the 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.Authoritative and cutting-edge, HIV Protocols, Third Edition is a great resource for HIV research, and contains many significant tips and tricks rarely mentioned in standard protocols.

Spis treści

Part I HIV-1 Replication - Early Events1. Quantifying CD4/CCR5 Usage Efficiency of HIV-1 ENV Using the Affinofile System
Nicholas E. Webb and Benhur Lee

2. Measuring T Cell-to-T Cell HIV-1 Transfer, Viral Fusion, and Infection Using Flow Cytometry
Natasha D. Durham and Benjamin K. Chen

3. HIV-1 Capsid Stabilization Assay
Thomas Fricke and Felipe Diaz-Griffero

4. Detection and Tracking of Dual-Labeled HIV Particles Using Wide-Field Live Cell Imaging to Follow Viral Core Integrity
Joao I. Mamede and Thomas J. Hope

5. HIV-1 Reverse Transcriptase Based Assay to Determine Cellular dNTP Concentrations
Joseph A. Hollenbaugh and Baek Kim

6. Rapid Determination of HIV-1 Mutant Frequencies and Mutation Spectra Using an mCherry/EGFP Dual Reporter Viral Vector
Jonathan M.O. Rawson, Christine L. Clouser, and Louis M. Mansky

Part II HIV-1 RNA Structure and RNA-Protein Interactions

7. Novel Biochemical Tools for Probing HIV RNA Structure
Jason W. Rausch, Joanna Sztuba-Solinska, Sabrina Lusvarghi, and Stuart F. J. Le Grice

8. Analysis of HIV-1 Gag-RNA Interactions in Cells and Virions by CLIP-seq
Sebla B. Kutluay and Paul D. Bieniasz

9. Isolation of Cognate Cellular and Viral Ribonucleoprotein Complexes of HIV-1 RNA Applicable to Proteomic Discovery and Molecular Investigations
Deepali Singh, Ioana Boeras, Gatikrushna Singh, and Kathleen Boris-Lawrie

Part III HIV-1 Replication - Post-Integrational Events

10. Methods for the Analyses of Inhibitor-Induced Aberrant Multimerization of HIV-1 Integrase
Jacques J. Kessl, Amit Sharma, and Mamuka Kvaratskhelia

11. Quantification of HIV-1 Gag Localization Within Virus Producer Cells
Annalena La Porte and Ganjam V. Kalpana

12. Methods to Study Determinants for Membrane Targeting of HIV-1 Gag In Vitro
Gabrielle C. Todd and Akira Ono

Part IV Studying HIV-1 Replication and Pathogenesis in Animal Models

13. Visualizing the Behavior of HIV-Infected T Cells In Vivo Using Multiphoton Intravital Microscopy
Radwa Sharaf, Thorsten R. Mempel, and Thomas T. Murooka

14. Modeling HIV-1 Mucosal Transmission and Prevention in Humanized Mice
Milena Veselinovic, Paige Charlins, and Ramesh Akkina

15. High Throughput Humanized Mouse Models for Evaluation of HIV-1 Therapeutics and Pathogenesis
Tynisha Thomas, Kieran Seay, Jian Hua Zheng, Cong Zhang, Christina Ochsenbauer, John C. Kappes, and Harris Goldstein

Part V Tools to Study HIV-1 Latency and Pathogenesis

16. Measuring the Frequency of Latent HIV-1 in Resting CD4⁺ T Cells Using a Limiting Dilution Co-Culture Assay
Gregory M. Laird, Daniel I.S. Rosenbloom, Jun Lai, Robert F. Siliciano, and Janet D. Siliciano

17. LGIT In Vitro Latency Model in Primary and T Cell Lines to Test HIV-1 Reactivation Compounds
Ulrike Jung, Mayumi Takahashi, John J. Rossi, and John C. Burnett

18. Improved Methods to Detect Low Levels of HIV Using Antibody-Based Technologies
Eliseo A. Eugenin and Joan W. Berman

19. Analysis of ABCA1 and Cholesterol Efflux in HIV-Infected Cells
Nigora Mukhamedova, Beda Brichacek, Christina Darwish, Anastas Popratiloff, Dmitri Sviridov, and Michael Bukrinsky

20. The Proteomic Characterization of Plasma or Serum from HIV-Infected Patients
Nicole A. Haverland, Lance M. Villeneuve, Pawel Ciborowski, and Howard S. Fox

21. Proteomic Characterization of Exosomes from HIV-1-Infected Cells
Ming Li and Bharat Ramratnam

Part VI NeuroAIDS

22. Detecting HIV-1 Tat in Cell Culture Supernatants by ELISA or Western Blot
Fabienne Rayne, Solène Debaisieux, Annie Tu, Christophe Chopard, Petra Tryoen, and Bruno Beaumelle

23. Protocol for Detection of HIV-Tat Protein in Cerebrospinal Fluid by a Sandwich Enzyme Linked Immunosorbent Assay
Tory P. Johnson and Avindra Nath

24. Measuring the Uptake and Transactivation Function of HIV-1 Tat Protein in a Trans-Cellular Co-Cultivation Set Up
Arthur P. Ruiz and Vinayaka R. Prasad

25. Evaluating the Role of Viral Proteins in HIV-Mediated Neurotoxicity Using Primary Human Neuronal Cultures
Vasudev R. Rao, Eliseo A. Eugenin, and Vinayaka R. Prasad