This collection of cutting-edge techniques is 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 studying 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.

Table of Contents:
Part I. 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 Part II. 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): A Field Approach to Therapeutic Monitoring Quantitation of HIV-1 RNA in Genital Secretions Detection of Nucleic Acids in Cells and Tissues by In Situ Polymerase Chain Reaction Omar Basgasra Determination of the Promoter Activity of HIV-1 Using the Chloramphenical 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 Ppriming as a Rapid Screen for Known Mutations Quantitation of HIV-1 Entry Cofactor Expression Retrovirus-Mediated Grene Transduction of SupT1 Cells Cloning of HIV Single-Strand DNA Binding Protein from Human Lymphocyte Lamda gt11 Expression Library with P32-Oligomers Telomeric Terminal Restriction Fragment (TRF) Part III. 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): Antibody Dilution Method Neutralization of HIV-1 Infection of Human Peripheral Blood Mononuclear Cells (PBMC): Infectivity Reduction Method Collection and Processing of Mucosal Secretions from HIV-1 Infected Women Collection and Processing of Mucosal Secretions from Mice Part IV. 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 Cell Populations using TUNEL Index.