HIV Protocols

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.