Library Science

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.  Ed. Michael, Nelson ; Kim, Jerome H.

Table of Contents

Part I
1. Virology Isolation and Expansion of HIV from Cells and Body Fluids by Coculture
   James R. Lane
2. Quantitative HIV Culture
   James R. Lane
3. Quantitation of Cell-Free HIV by Reverse Transcriptase Activity
   James R. Lane
4. Determination of Syncytium-Inducing Phenotype of Primary HIV-1 Isolates Using MT-2 Cells
   Mark K. Louder and John R. Mascola
5. Determination of HIV-1 Chemokine Coreceptor Tropism Using Transduced Human Osteosarcoma (HOS) Cells
   Mark K. Louder and John R. Mascola
6. Use of Luciferase Reporter Viruses for Studying HIV Entry
   Rong Liu, Benjamin Chen, and Nathaniel R. Landau
7. A Cell-Cell Fusion Assay to Monitor HIV-1 Env Interactions with Chemokine Receptors
   Aimee L. Edinger and Robert W. Doms
8. Methods of Culturing HIV-1 from Semen
   Bruce L. Gilliam

Part II

9. Molecular Biology Detection of HIV-1 Nucleic Acids by Southern Blotting
   Richard A. McDonald
10. Detection of HIV-1 Nucleic Acids by Northern Blotting
    Richard A. McDonald
11. Rapid Identification of Cloned HIV-1 Fragments
    Wei Dong, Anindita Kar Roy, and Yen Li
12. PCR Amplification and Cloning of Virtually Full-Length HIV-1 Provirus
    Mika Salminen
13. Quantitation of HIV-1gag DNA and RNA from Single Frozen Cell Pellets
    Maryanne Vahey, Sandra C. Barrick, and Martin Nau
14. Quantitative PCR for HIV-1 Proviral DNA
    D . Zella and P. Secchiero
15. Diagnosis and Direct Automated Sequencing of HIV-1 From Dried Blood Spots (DBS) Collected on Filter Paper
    Sharon A. Cassol, Stanley Read, Bruce G. Weniger, Richard Pilon, Barbara Leung, and Theresa Mo
16. Quantification of HIV-1 RNA in Dried Plasma Spots (DPS): A Field Approach to Therapeutic Monitoring
    Sharon A. Cassol, Francisco Diaz-Mitoma, Richrd Pilon, Michelle James, and D. William Cameron
17. Quantitation of HIV-1 RNA in Genital Secretions
    Bruce L. Gilliam
18. Detection of Nucleic Acids in Cells and Tissues by In Situ Polymerase Chain Reaction
    Omar Basgasra, Lisa E. Bobroski, Mohammad Amjad, Matthew Memdi, and Maureen V. Abbey
19. Determination of the Promoter Activity of HIV-1 Using the Chloramphenical Acetyltransferase Reporter Gene Assay
    Loretta Tse and Nelson L. Michael
20. In Vitro Techniques for Studies of HIV-1 Promoter Activity
    Fabio Romerio and David M. Margolis
21. Detection of Polymorphisms in the HIV-1 Coreceptor CCR5 Using Single-Strand Conformation Polymorphism
    Maureen P. Martin and Mary Carrington
22. Sequence-Specific Ppriming as a Rapid Screen for Known Mutations
    Mary Carrington
23. Quantitation of HIV-1 Entry Cofactor Expression
    James L. Riley and Richard G. Carroll
24. Retrovirus-Mediated Grene Transduction of SupT1 Cells
    Jerome K. Kim and J. Lin
25. Cloning of HIV Single-Strand DNA Binding Protein from Human Lymphocyte Lamda gt11 Expression Library with P32-Oligomers
    S. J. Hur and D. Lai
26. Telomeric Terminal Restriction Fragment (TRF)
    Sumesh Kaushal

Part III

27. Humoral Immunology Serologic Analysis by Enzyme-Linked Immunosorbent Assay (ELISA)
    Melissa Krider and Lawrence D. Loomis-Price
28. Quantitative Immunoblotting with Fusion Proteins (Loomis-Price )
    Margaret M. Mitchell and Lawrence D.
29. Analysis of Antibody Interactions with HIV-1 Envelope Expressed on the Surface of Acutely Infected H9 Cells
    Thomas C. VanCott and Paul L. Hallberg
30. Analysis of Antibody-Antigen Interactions Using Surface Plasmon Resonance
    Thomas C. VanCott
31. Measurement of HIV-1 Specific and Total Antibody Secreting Cells by ELISPOT
    Frederik W. van Ginkel, John L. VanCott, Robert Kaminski, and Thomas C. VanCott
32. Linear Epitope Mapping by the PEPSCAN Method
    Lawrence D. Loomis-Price
33. Neutralization of HIV-1 Infection of Human Peripheral Blood, Mononuclear Cells (PBMC): Antibody Dilution Method
    John R. Mascola
34. Neutralization of HIV-1 Infection of Human Peripheral Blood, Mononuclear Cells (PBMC): Infectivity Reduction Method
    John R. Mascola
35. Collection and Processing of Mucosal Secretions from HIV-1 Infected Women
    Andrew W. Artenstein and Thomas C. VanCott
36. Collection and Processing of Mucosal Secretions from Mice
    Robert W. Kamonski and Thomas C. VanCott

Part IV

37. Cellular Immunology Lymphocyte Proliferation Assay
    Karl V. Sitz and Deborah L. Birx
38. HIV-1 Specific Cytotoxic T-Cell Assays Josephine H. (osephine H. Cox )
    Cox HIV-1-Specific Antibody-Dependent Cellular Cytotoxicity (ADCC) J
39. Evaluation of Natural Killer Cell Activity
    Josephine H. Cox
40. Generation and Expansion of HIV-1 CD4+ Antigen-Specific T Cell Lines
    Silvia Ratto-Kim
41. Immunophenotyping and Assessment of Cell Function by Three-Color Flow Cytometry of Peripheral Blood Lymphocytes
    Stephen P. Perfetto and Gilbert McCrary
42. Detection of Apoptosis in HIV-Infected Cell Populations using TUNEL
    Patrick J. Blair and Stephen P. Perfetto

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