HIV protocols / edited by Vinayaka R. Prasad and Ganjam V. Kalpana. — 2nd ed. — New York : Humana Press, c2009. – (58.17/M592/v.485)

Contents

    Contents
    
    Preface
    Contributors
    SECTION I. PREPARATION OF VIRUS PARTICLES AND THEIR ANALYSIS
    1 Methods for Viral RNA Isolation and PCR Amplification for Sequencing of Near Full-Length HIV-1 Genomes
    2 Purification ofHIV-1 Virions by Subtilisin Digestion or CD45 Immunoaffinity Depletion for Biochemical Studies
    3 Calculating HIV-1 Infectious Titre Using a Virtual TCIDs0 Method
    SECTION II. METHODS TO STUDY HIV-1 REPLICATION
    SUBSECTION A. EARLY EVENTS
    4 Cell-Free Assays for HIV-1 Uncoating
    5 Real-Time PCR Analysis of HIV-1 Replication Postentry Events
    6 Analysis of 2-LTR Circle Junctions of Viral DNA in Infected Cells
    7 HIV-1 Recombination: An Experimental Assay and a Phylogenetic Approach
    8 Methods of Preparation and Analysis of Intracellular Reverse Transcription Complexes
    9 Analysis of Viral and Cellular Proteins in HIV-1 Reverse Transcription Complexes by Co-immunoprecipitation
    10 Isolation and Analysis of HIV-1 Preintegration Complexes
    11 Bisarsenical Labeling of HIV-1 for Real-Time Fluorescence Microscopy
    SUBSECTION B. LATE EVENTS
    12 Methods for the Study of HIV-1 Assembly
    13 Assembly of Immature HIV-1 Capsids Using a Cell-Free System
    14 Preparation of Recombinant Hiv-1 Gag Protein and Assembly of Virus-Like Particles In Vitro
    15 Methods for the Analysis of HIV-1 Nucleocapsid Protein Interactions with Oligonucleotides
    16 Methods for Analysis of Incorporation and Annealing of tRNALys in HIV- 1
    SECTION III. SPECIALIZED APPROACHES TO STUDY HIV-1 BIOLOGY AND PATHOGENESIS
    17 Somatic Cell Genetic Analyses to Identify HIV-1 Host Restriction Factors
    18 Rapid， Controlled and Intensive Lentiviral Vector-Based RNAi
    19 Reverse Two-Hybrid Screening to Analyze Protein-Protein Interaction of HIV-1 Viral and Cellular Proteins
    20 Methods to Study Monocyte Migration Induced by HIV-Infected Cells
    21 Novel Mouse Models for Understanding HIV-1 Pathogenesis
    SECTION IV. IMMUNOLOGICAL STUDIES OF HIV
    SUBSECTION A. MUCOSAL IMMUNOLOGY
    22 Mucosal Antibody Responses to HIV
    23 Isolating Mucosal Lymphocytes from Biopsy Tissue for Cellular Immunology Assays
    SUBSECTIOM B. MEASURING T CELL RESPONSES VIA FLOW CYTOMETRY
    24 Quantifying HIV-1-Specific CD8+ T-Cell Responses Using ELISPOT and Cytokine Flow Cytometry
    25 Multiparameter Flow Cytometry Monitoring ofT Cell Responses
    SUBSECTION C. ANTIVIRAL RESPONSES
    26 Measuring HIV Neutralization in a Luciferase Reporter Gene Assay
    27 Assessing the Antiviral Activity of HIV-1-Specific Cytotoxic T Lymphocytes
    28 Methods for Quantitating Antigen-Specific T Cell Responses Using Functional Assays in Rhesus Macaques
    SECTION V. DRUG RESISTANT VIRUSES AND VIRAL EVOLUTION
    29 Isolation of Drug-Resistant Mutant HIV Variants Using Tissue Culture Drug Selection
    30 Virus Evolution as a Tool to Study HIV-1 Biology
    Index