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This book details the development of methods and models to study the HIV-1 viral reservoir with the ultimate goal of achieving a functional cure of HIV infection. Chapters are divided into six parts covering cell lines, in vitro and ex vivo primary cell models of persistent infection, in vitro and ex vivo tissue-derived models, infected animal models human immune cells, methods of detection and analysis of the reservoir, and current approaches to achieve either a functional cure or cART-free long-term remission. Written in the format of the highly successful Methods in Molecular Biology series, each chapter includes an introduction to the topic, lists necessary materials and reagents, includes tips on troubleshooting and known pitfalls, and step-by-step, readily reproducible protocols.
Authoritative and cutting-edge, HIV Reservoirs: Methods and Protocols provides a comprehensive, updated collection of state-of-art methodologies and models to tackle the HIV-1 viral reservoir.
List of contents
Jurkat-derived (J-Lat, J1.1 and Jurkat E4) and CEM-derived T cell lines (8E5, ACH-2) as Models of Reversible Proviral Latency.- U1 and OM10.1. Myeloid Cell Lines as Surrogate Models Of Reversible Proviral Latency.- An In Vitro System to Model the Establishment and Reactivation of HIV-1 Latencyin primary human CD4+ T cells.- The cultured TCM model of HIV latency.- A Reliable Primary Cell Model for HIV latency: The QUECEL (Quiescent Effector Cell Latency) Method.- TGF-b signaling supports HIV latency in a Memory CD4+ T cell based in vitro model.p Flow Cytometry Sorting of Memory CCR6+CD4+ T-Cells for HIV Reservoirs Quantification.- Human monocyte-derived macrophages (MDM) - Model 1 (GM-CSF).- Human monocyte-derived macrophages (MDM) - Model 2.- Modeling HIV latency in astrocytes with the human neural progenitor cell line HNSC.100.- Hematopoietic stem and progenitor cells (HSPCs).- HIV reservoirs: Modeling, Quantification and Approaches to a Cure.- More than a gender issue: Testis as a distinctive HIV reservoir and its implication for viral eradication.- Experimental Models to Study HIV Latency Reversal from Male Genital Myeloid Cells.- Decidua basalis: An ex vivo Model to Study HIV-1 Infection during Pregnancy and Beyond.- HIV Reservoirs: Modeling, Quantification, and Approaches to a Cure.- Human Hematopoietic Stem Cell (HSC)-Engrafted NSG Mice for HIV Latency Research.- Inducing long-term HIV-1 latency in the TKO-BLT mouse model.- In situ multiplexing to identify, quantify, and phenotype the HIV-1/SIV reservoir within lymphoid tissue.- Single-cell multiparametric analysis of rare HIV-infected cells identified by duplexed RNAflow-FISH.- Ex vivo differentiation of resting CD4+ T lymphocytes enhances detection of replication competent HIV-1 in viral outgrowth assays.- Quantitative Viral Outgrowth Assay to Measure the functional SIV Reservoir in Myeloid Cells.- Near-full-length single-genome HIV-1 DNA sequencing.- TILDA: Tat/rev Induced Limiting Dilution Assay.- Latency Reversal and Clearance of Persistent HIV Infection.- Cure and long-term remission strategies.- Pathways Toward a Functional HIV-1 Cure: Balancing Promise and Perils for CRISPR Therapy.
