Follows the step-by-step life cycle of a retrovirus
Provides state-of-the-art methodological protocols from world leaders in human retrovirology
Includes expert notes on troubleshooting and avoiding known pitfalls
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.
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, TRIM5?
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 Tranillegalscription
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, Tranillegalscription, 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. SIVSM/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
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.
Virion Attachment & Entry: HIV gp120 Env Biotinylation, gp120 Env, or Integrin Ligand Binding Assay.- CryoEM Analysis of Capsid Assembly and Structural Changes Upon Interactions with a Host Restriction Factor, TRIM5 .- The Fate of HIV-1 Capsid: A Biochemical Assay for HIV-1 Uncoating.- The Cyclosporin A Washout Assay to Detect HIV-1 Uncoating in Infected Cells.- Imaging HIV-1 Nuclear Pre-Integration Complexes.- HIV-1 Reverse Tranillegalscription.- RNase H: Specificity, Mechanisms of Action, and Antiviral Target.- HIV-1 Chromatin, Tranillegalscription, and the Regulatory Protein Tat.- HIV-1 Rev Function and RNA Nuclear-Cytoplasmic Export.- HIV-1 Accessory Proteins: Nef.- HIV-1 Accessory Proteins: VpR.- HIV-1 Accessory Proteins: Vpu and Vif.- SIVSM/HIV-2 Vpx Proteins : Function and Uses in the Infection of Primary Myeloid Cells.- Imaging of HIV Assembly and Release.- HIV-1 Isolation from Infected Peripheral Blood Mononuclear Cells.- Determination of HIV-1 Co-Receptor Usage.- The Macrophage and HIV: Basic Concepts and Methodologies.- HIV Infection of Dendritic Cells.- Histocultures (Tissue Explants) in Human Retrovirology.- Single Copy Quantification of HIV-1 in Clinical Samples.- Quantification of Total HIV1-DNA in Peripheral Blood Mononuclear Cells.- HIV-1 Based Lentiviral Vectors.- Quantification of miRNA by Poly(A)-RT-qPCR Arrays and Verification of Target Sites in HIV-1 Using a One-LTR Infectious Molecular Clone.- Investigating Human T Cell Lymphotropic Retrovirus (HTLV) Tax Function with Molecular and Immunophenotypic Techniques.- Proviral Load Determination of HTLV-1 and HTLV-2 in Patients' Peripheral Blood Mononuclear Cells by Real-Time PCR.- Quantitative Analysis of HumanT-Lymphotropic Virus Type 1 (HTLV-1) Gene Expression Using Nucleo-Cytoplasmic Fractionation and Splice Junction-Specific Real Time RT-PCR (qRT-PCR).
Inhaltsverzeichnis
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, TRIM5a
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 Tranillegalscription
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, Tranillegalscription, 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. SIVSM/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
Klappentext
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.n nComprehensive 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.
Follows the step-by-step life cycle of a retrovirus
Provides state-of-the-art methodological protocols from world leaders in human retrovirology
Includes expert notes on troubleshooting and avoiding known pitfalls
Includes supplementary material: sn.pub/extras
