Virology -- Isolation and Expansion of HIV from Cells and Body Fluids by Coculture -- Quantitative HIV Culture -- Quantitation of Cell-Free HIV by Reverse Transcriptase Activity -- Determination of Syncytium-Inducing Phenotype of Primary HIV-1 Isolates Using MT-2 cells -- Determination of HIV-1 Chemokine Coreceptor Tropism Using Transduced Human Osteosarcoma (HOS) Cells -- Use of Luciferase Reporter Viruses for Studying HIV Entry -- A Cell-Cell Fusion Assay to Monitor HIV-1 Env Interactions with Chemokine Receptors -- Methods of Culturing HIV-1 from Semen -- Molecular Biology -- Detection of HIV-1 Nucleic Acids by Southern Blotting -- Detection of HIV-1 Nucleic Acids by Northern Blotting -- Rapid Identification of Cloned HIV-1 Fragments -- PCR Amplification and Cloning of Virtually Full-Length HIV-1 Provirus -- Quantitation of HIV-1gag DNA and RNA From Single Frozen Cell Pellets -- Quantitative PCR for HIV-1 Proviral DNA -- Diagnosis and Direct Automated Sequencing of HIV-1 From Dried Blood Spots (DBS) Collected on Filter Paper -- Quantification of HIV-1 RNA in Dried Plasma Spots (DPS) -- Quantitation of HIV-1 RNA in Genital Secretions -- Detection of Nucleic Acids in Cells and Tissues by In Situ Polymerase Chain Reaction -- Determination of the Promoter Activity of HIV-1 Using the Chloramphenicol Acetyltransferase Reporter Gene Assay -- In Vitro Techniques for Studies of HIV-1 Promoter Activity -- Detection of Polymorphisms in the HIV-1 Coreceptor CCR5 Using Single-Strand Conformation Polymorphism -- Sequence-Specific Priming as a Rapid Screen for Known Mutations -- Quantitation of HIV-1 Entry Cofactor Expression -- Retrovirus-Mediated Gene Transduction of SupT1 Cells -- Cloning of HIV Single-Strand DNA Binding Protein from Human Lymphocyte Lambda gt11 Expression Library with [32P]-Oligomers -- Telomeric Terminal Restriction Fragment (TRF) -- Humoral Immunology -- Serologic Analysis by Enzyme-Linked Immunosorbent Assay (ELISA) -- Quantitative Immunoblotting with Fusion Proteins -- Analysis of Antibody Interactions with HIV-1 Envelope Expressed on the Surface of Acutely Infected H9 Cells -- Analysis of Antibody-Antigen Interactions Using Surface Plasmon Resonance -- Measurement of HIV-1 Specific and Total Antibody Secreting Cells by ELISPOT -- Linear Epitope Mapping by the PEPSCAN Method -- Neutralization of HIV-1 Infection of Human Peripheral Blood Mononuclear Cells (PBMC) -- Neutralization of HIV-1 Infection of Human Peripheral Blood Mononuclear Cells (PBMC) -- Collection and Processing of Mucosal Secretions from HIV-1 Infected Women -- Collection and Processing of Mucosal Secretions from Mice -- Cellular Immunology -- Lymphocyte Proliferation Assay -- HIV-1-Specific Cytotoxic T-Cell Assays -- HIV-1-Specific Antibody-Dependent Cellular Cytotoxicity (ADCC) -- Evaluation of Natural Killer Cell Activity -- Generation and Expansion of HIV-1 CD4+ Antigen-Specific T Cell Lines -- Immunophenotyping and Assessment of Cell Function by Three-Color Flow Cytometry of Peripheral Blood Lymphocytes -- Detection of Apoptosis in HIV-Infected Ceil Populations using TUNEL.

In HIV Protocols, Nelson Michael and Jerome Kim have organized a collection of cutting-edge techniques essential for studying the molecular biology, virology, and immunology of the HIV virus. The methods cover a broad range of research interests, including quantitation of viral genomes, HIV promoter function, B-cell epitope mapping, viral coreceptor usage, and measurements of T-cell function. Special emphasis is accorded to the study of those viral and host immune responses to infection that will be critical to the design of effective preventive vaccines. HIV Protocols is the first HIV methods book to concentrate equally on virology, molecular biology, and immunology, as well as to incorporate methods on chemokine receptor structure and function. Comprehensive and state-of-the-art, the techniques detailed here provide an indispensable framework for unraveling the molecular and immunological aspects of HIV infection.