Human Retroviruses: Methods and Protocols

ISBN-13: 9781493962587 / Angielski / Miękka / 2016 / 345 str.

Elisa Vicenzi; Guido Poli

Opis

Human Retroviruses: Methods and Protocols collects key experimental protocols that have provided the basis of the major discoveries of the field. Split into five sections, this detailed volume covers mapping of the HIV life cycle, isolation, co-receptor use, and cell tropism of HIV-1, in vivo quantification of HIV-1, biological aspects of HIV-1, as well as HTLVs. Some articles explore "assay and function of accessory genes," largely involving the interface between retroviral and host factors, the extracellular role of Tat and Tax, resembling the function of cytokines, and the biotechnological exploitation of HIV as lentiviral vector to carry foreign genes with therapeutic value. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Comprehensive and authoritative, Human Retroviruses: Methods and Protocols provides state-of-art methodological protocols from world leaders in human retrovirology, essential for any lab working this vital field.

Spis treści

Part I: Mapping the HIV Life Cycle

 

1. Virion Attachment & Entry: HIV gp120 Env Biotinylation, gp120 Env, or Integrin Ligand Binding Assay

            Claudia Cicala and James Arthos

 

2. CryoEM Analysis of Capsid Assembly and Structural Changes Upon Interactions with a Host Restriction Factor, TRIM5α

            Gongpu Zhao and Peijun Zhang

 

3. The Fate of HIV-1 Capsid: A Biochemical Assay for HIV-1 Uncoating

            Yang Yang, Jeremy Luban, and Felipe Diaz-Griffero

 

4. The Cyclosporin A Washout Assay to Detect HIV-1 Uncoating in Infected Cells

            Thomas Hope

 

5. Imaging HIV-1 Nuclear Pre-Integration Complexes

            Anna Cereseto and Mauro Giacca

 

6. HIV-1 Reverse Transcription

            Andrea Cimarelli and Jean-Luc Darlix

 

7. RNase H: Specificity, Mechanisms of Action, and Antiviral Target

            Karin Moelling, Felix Broecker, and John E. Kerrigan

 

8. HIV-1 Chromatin, Transcription, and the Regulatory Protein Tat

            Laurence Colin, Eric Verdin, and Carine Van Lint

 

9. HIV-1 Rev Function and RNA Nuclear-Cytoplasmic Export

            Alan Cochrane

 

10. HIV-1 Accessory Proteins: Nef

            Anke Heigele, Daniel Sauter, Jan Münch, and Frank Kirchhoff

 

11. HIV-1 Accessory Proteins: VpR

            Richard Y. Zhao and Michael I. Bukrinsky

 

12. HIV-1 Accessory Proteins: Vpu and Vif

            Amy Andrew and Klaus Strebel

 

13. SIV_SM/HIV-2 Vpx Proteins : Function and Uses in the Infection of Primary Myeloid Cells

            Gregory Berger and Andrea Cimarelli

 

14. Imaging of HIV Assembly and Release

            Barbara Müller and Jacomine Krijnse-Locker

 

Part II: Isolation, Co-Receptor Use, and Cell Tropism of HIV-1

 

15. HIV-1 Isolation from Infected Peripheral Blood Mononuclear Cells

            Stefania Dispinseri, Elisa Saba, Elisa Vicenzi, Neeltje A. Kootstra, Hanneke Schuitemaker, and Gabriella Scarlatti

 

16. Determination of HIV-1 Co-Receptor Usage

            Mariangela Cavarelli and Gabriella Scarlatti

 

17. The Macrophage and HIV: Basic Concepts and Methodologies

            Suzanne Gartner

 

18. HIV Infection of Dendritic Cells

            Najla Nasr, Andrew Harman, Stuart Turville and Anthony L. Cunningham

 

19. Histocultures (Tissue Explants) in Human Retrovirology

            Anush Arakelyan, Wendy Fitzgerald, Jean-Charles Grivel, Christophe Vanpouille, and Leonid Margolis

 

Part III: In Vivo Quantification of HIV-1

 

20. Single Copy Quantification of HIV-1 in Clinical Samples

            Ann Wiegand and Frank Maldarelli

 

21. Quantification of Total HIV1-DNA in Peripheral Blood Mononuclear Cells

            Christine Rouzioux, Adeline Mélard, and Véronique Avéttand-Fénoël

 

Part IV: Biotechnological Aspects of HIV-1

 

22. HIV-1 Based Lentiviral Vectors

            Ying Poi Liu and Ben Berkhout

 

23. Quantification of miRNA by Poly(A)-RT-qPCR Arrays and Verification of Target Sites in HIV-1 Using a One-LTR Infectious Molecular Clone

            Zachary A. Klase, Laurent Houzet, and Kuan-Teh Jeang

 

Part V: HTLVs

 

24. Investigating Human T Cell Lymphotropic Retrovirus (HTLV) Tax Function with Molecular and Immunophenotypic Techniques

            Greta Forlani, Roberto S. Accolla, and Giovanna Tosi

 

25. Proviral Load Determination of HTLV-1 and HTLV-2 in Patients’ Peripheral Blood Mononuclear Cells by Real-Time PCR

            Claudio Casoli, Elisabetta Pilotti, and Umberto Bertazzoni

 

26. Quantitative Analysis of Human T-Lymphotropic  Virus Type 1 (HTLV-1) Gene Expression Using Nucleo-Cytoplasmic Fractionation and Splice Junction-Specific Real Time RT-PCR (qRT-PCR)

            Ilaria Cavallari, Francesca Rende, and Vincenzo Ciminale