HIV Protocols

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.

Includes cutting-edge methods and protocols Provides step-by-step detail essential for reproducible results Contains key notes and implementation advice from the experts

Contenu

Quantifying CD4/CCR5 Usage Efficiency of HIV-1 ENV Using the Affinofile System.- Measuring T Cell-to-T Cell HIV-1 Transfer, Viral Fusion, and Infection Using Flow Cytometry.- HIV-1 Capsid Stabilization Assay.- Detection and Tracking of Dual-Labeled HIV Particles Using Wide-Field Live Cell Imaging to Follow Viral Core Integrity.- HIV-1 Reverse Transcriptase Based Assay to Determine Cellular dNTP Concentrations.- Rapid Determination of HIV-1 Mutant Frequencies and Mutation Spectra Using an mCherry/EGFP Dual Reporter Viral Vector.- Novel Biochemical Tools for Probing HIV RNA Structure.- Analysis of HIV-1 Gag-RNA Interactions in Cells and Virions by CLIP-seq.- Isolation of Cognate Cellular and Viral Ribonucleoprotein Complexes of HIV-1 RNA Applicable to Proteomic Discovery and Molecular Investigations.- Methods for the Analyses of Inhibitor-Induced Aberrant Multimerization of HIV-1 Integrase.- Quantification of HIV-1 Gag Localization Within Virus Producer Cells.- Methods to Study Determinants for Membrane Targeting of HIV-1 Gag In Vitro.- Visualizing the Behavior of HIV-Infected T Cells In Vivo Using Multiphoton Intravital Microscopy.- Modeling HIV-1 Mucosal Transmission and Prevention in Humanized Mice.- High Throughput Humanized Mouse Models for Evaluation of HIV-1 Therapeutics and Pathogenesis.- Measuring the Frequency of Latent HIV-1 in Resting CD4 + T Cells Using a Limiting Dilution Co-Culture Assay.- LGIT In Vitro Latency Model in Primary and T Cell Lines to Test HIV-1 Reactivation Compounds.- Improved Methods to Detect Low Levels of HIV Using Antibody-Based Technologies.- Analysis of ABCA1 and Cholesterol Efflux in HIV-Infected Cells.- The Proteomic Characterization of Plasma or Serum from HIV-Infected Patients.- Proteomic Characterization of Exosomes from HIV-1-Infected Cells.- Detecting HIV-1 Tat in Cell Culture Supernatants by ELISA or Western Blot.- Protocol for Detection of HIV-Tat Protein in Cerebrospinal Fluid by a Sandwich Enzyme Linked Immunosorbent Assay.- Measuring the Uptake and Transactivation Function of HIV-1 Tat Protein in a Trans-Cellular Co-Cultivation Set Up.- Evaluating the Role of Viral Proteins in HIV-Mediated Neurotoxicity Using Primary Human Neuronal Cultures.