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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 15: High-Throughput Humanized Mouse Models for Evaluation of HIV-1 Therapeutics and Pathogenesis
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Chapter title
High-Throughput Humanized Mouse Models for Evaluation of HIV-1 Therapeutics and Pathogenesis
Chapter number 15
Book title
HIV Protocols
Published in
Methods in molecular biology, January 2016
DOI 10.1007/978-1-4939-3046-3_15
Pubmed ID
Book ISBNs
978-1-4939-3045-6, 978-1-4939-3046-3

Tynisha Thomas, Kieran Seay, Jian Hua Zheng, Cong Zhang, Christina Ochsenbauer, John C. Kappes, Harris Goldstein


Mice cannot be used as a model to evaluate HIV-1 therapeutics because they do not become infected by HIV-1 due to structural differences between several human and mouse proteins required for HIV-1 replication. This has limited their use for in vivo assessment of anti-HIV-1 therapeutics and the mechanism by which cofactors, such as illicit drug use accelerate HIV-1 replication and disease course in substance abusers. Here, we describe the development and application of two in vivo humanized mouse models that are highly sensitive and useful models for the in vivo evaluation of candidate anti-HIV therapeutics. The first model, hu-spl-PBMC-NSG mice, uses NOD-SCID IL2rγ(-/-) (NSG) mice intrasplenically injected with human peripheral blood mononuclear cells (PBMC) which develop productive splenic HIV-1 infection after intrasplenic inoculation with a replication-competent HIV-1 expressing Renilla reniformis luciferase (HIV-LucR) and enables investigators to use bioluminescence to visualize and quantitate the temporal effects of therapeutics on HIV-1 infection. The second model, hCD4/R5/cT1 mice, consists of transgenic mice carrying human CD4, CCR5 and cyclin T1 genes, which enables murine CD4-expressing cells to support HIV-1 entry, Tat-mediated LTR transcription and consequently develop productive infection. The hCD4/R5/cT1 mice develop disseminated infection of tissues including the spleen, small intestine, lymph nodes and lungs after intravenous injection with HIV-1-LucR. Because these mice can be infected with HIV-LucR expressing transmitted/founder and clade A/E and C Envs, these mouse models can also be used to evaluate the in vivo efficacy of broadly neutralizing antibodies and antibodies induced by candidate HIV-1 vaccines. Furthermore, because hCD4/R5/cT1 mice can be infected by vaginal inoculation with replication-competent HIV-1 expressing NanoLuc (HIV-nLucR)-, this mouse model can be used to evaluate the mechanisms by which substance abuse and other factors enhance mucosal transmission of HIV-1.

Mendeley readers

The data shown below were compiled from readership statistics for 10 Mendeley readers of this research output. Click here to see the associated Mendeley record.

Geographical breakdown

Country Count As %
Canada 1 10%
Unknown 9 90%

Demographic breakdown

Readers by professional status Count As %
Student > Master 3 30%
Professor 1 10%
Researcher 1 10%
Student > Ph. D. Student 1 10%
Other 1 10%
Other 1 10%
Unknown 2 20%
Readers by discipline Count As %
Agricultural and Biological Sciences 3 30%
Biochemistry, Genetics and Molecular Biology 2 20%
Nursing and Health Professions 1 10%
Immunology and Microbiology 1 10%
Medicine and Dentistry 1 10%
Other 0 0%
Unknown 2 20%