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HIV Protocols

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Cover of 'HIV Protocols'

Table of Contents

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    Book Overview
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    Chapter 1 Quantifying CD4/CCR5 Usage Efficiency of HIV-1 Env Using the Affinofile System
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    Chapter 2 Measuring T Cell-to-T Cell HIV-1 Transfer, Viral Fusion, and Infection Using Flow Cytometry
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    Chapter 3 HIV-1 Capsid Stabilization Assay
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    Chapter 4 Detection and Tracking of Dual-Labeled HIV Particles Using Wide-Field Live Cell Imaging to Follow Viral Core Integrity
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    Chapter 5 HIV-1 Reverse Transcriptase-Based Assay to Determine Cellular dNTP Concentrations
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    Chapter 6 Rapid Determination of HIV-1 Mutant Frequencies and Mutation Spectra Using an mCherry/EGFP Dual-Reporter Viral Vector
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    Chapter 7 Novel Biochemical Tools for Probing HIV RNA Structure
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    Chapter 8 Analysis of HIV-1 Gag-RNA Interactions in Cells and Virions by CLIP-seq.
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    Chapter 9 Isolation of Cognate Cellular and Viral Ribonucleoprotein Complexes of HIV-1 RNA Applicable to Proteomic Discovery and Molecular Investigations
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    Chapter 10 Methods for the Analyses of Inhibitor-Induced Aberrant Multimerization of HIV-1 Integrase
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    Chapter 11 Quantification of HIV-1 Gag Localization Within Virus Producer Cells
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    Chapter 12 Methods to Study Determinants for Membrane Targeting of HIV-1 Gag In Vitro
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    Chapter 13 Visualizing the Behavior of HIV-Infected T Cells In Vivo Using Multiphoton Intravital Microscopy
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    Chapter 14 Modeling HIV-1 Mucosal Transmission and Prevention in Humanized Mice
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    Chapter 15 High-Throughput Humanized Mouse Models for Evaluation of HIV-1 Therapeutics and Pathogenesis
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    Chapter 16 Measuring the Frequency of Latent HIV-1 in Resting CD4 + T Cells Using a Limiting Dilution Coculture Assay
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    Chapter 17 LGIT In Vitro Latency Model in Primary and T Cell Lines to Test HIV-1 Reactivation Compounds
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    Chapter 18 Improved Methods to Detect Low Levels of HIV Using Antibody-Based Technologies
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    Chapter 19 Analysis of ABCA1 and Cholesterol Efflux in HIV-Infected Cells
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    Chapter 20 The Proteomic Characterization of Plasma or Serum from HIV-Infected Patients
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    Chapter 21 Proteomic Characterization of Exosomes from HIV-1-Infected Cells
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    Chapter 22 Detecting HIV-1 Tat in Cell Culture Supernatants by ELISA or Western Blot
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    Chapter 23 Protocol for Detection of HIV-Tat Protein in Cerebrospinal Fluid by a Sandwich Enzyme-Linked Immunosorbent Assay
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    Chapter 24 Measuring the Uptake and Transactivation Function of HIV-1 Tat Protein in a Trans-cellular Cocultivation Setup.
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    Chapter 25 Evaluating the Role of Viral Proteins in HIV-Mediated Neurotoxicity Using Primary Human Neuronal Cultures.
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    Chapter 26 Erratum
Attention for Chapter 2: Measuring T Cell-to-T Cell HIV-1 Transfer, Viral Fusion, and Infection Using Flow Cytometry
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Chapter title
Measuring T Cell-to-T Cell HIV-1 Transfer, Viral Fusion, and Infection Using Flow Cytometry
Chapter number 2
Book title
HIV Protocols
Published in
Methods in molecular biology, January 2016
DOI 10.1007/978-1-4939-3046-3_2
Pubmed ID
Book ISBNs
978-1-4939-3045-6, 978-1-4939-3046-3
Authors

Natasha D. Durham, Benjamin K. Chen

Abstract

Direct T cell-to-T cell HIV-1 infection is a distinct mode of HIV-1 infection that requires physical contact between an HIV-1-infected "donor" cell and an uninfected, CD4-expressing "target" cell. In vitro studies indicate that HIV-1 cell-to-cell infection is much more efficient than infection by cell-free viral particles; however, the exact mechanisms of the enhanced efficiency of this infection pathway are still unclear. Several assays have been developed to study the mechanism of direct cell-to-cell HIV-1 transmission and to assess sensitivity to neutralizing antibodies and pharmacologic inhibitors. These assays are based on the coculture of donor and target cells. Here, we describe methods that utilize flow cytometry, which can discriminate donor and target cells and can assess different stages of entry and infection following cell-to-cell contact. HIV Gag-iGFP, a clone that makes fluorescent virus particles, can be used to measure cell-to-cell transfer of virus particles. HIV NL-GI, a clone that expresses GFP as an early gene, facilitates the measure of productive infection after cell-to-cell contact. Lastly, a variation of the β-lactamase (BlaM)-Vpr fusion assay can be used to measure the viral membrane fusion process after coculture of donor and target cells in a manner that is independent of cell-cell fusion. These assays can be performed in the presence of neutralizing antibodies/inhibitors to determine the 50 % inhibitory concentration (IC50) required to block infection specifically in the target cells.

Mendeley readers

The data shown below were compiled from readership statistics for 5 Mendeley readers of this research output. Click here to see the associated Mendeley record.

Geographical breakdown

Country Count As %
Unknown 5 100%

Demographic breakdown

Readers by professional status Count As %
Student > Ph. D. Student 1 20%
Professor > Associate Professor 1 20%
Other 1 20%
Student > Master 1 20%
Unknown 1 20%
Readers by discipline Count As %
Agricultural and Biological Sciences 2 40%
Immunology and Microbiology 2 40%
Unknown 1 20%