Pseudotyped Viruses

Preface
Contents
Editor and Contributors
Chapter 1: Pseudotyped Viruses
1.1 Introduction
1.2 The Vectors for Enveloped Viruses
1.2.1 Lentiviral Vectors for Enveloped Viruses
1.2.1.1 HIV Vector
1.2.1.2 Murine Leukemia Virus Vector
1.2.1.3 FIV Vector
1.2.2 The Vesicular Stomatitis Virus Vector
1.2.2.1 VSV Reverse Genetics
1.2.2.2 Recombinant VSV Vectors
1.3 Self-Assembled Constructed Pseudotyped Viruses
1.3.1 Self-Assembled Pseudotyped Virus for Enveloped Viruses
1.3.2 Self-Assembled Pseudotyped Virus for Non-enveloped Viruses
1.4 The Parameters and Conditions for Construction and Package of Pseudotyped Viruses
1.4.1 Viral Biological Characteristic Affects Pseudotyped Virus Formation and Titer
1.4.2 The Effects of Envelope Protein Expression
1.4.3 The Effects of Packaging System
1.4.4 Effects of Proteases
1.4.5 Selection of Cell Lines for Pseudovirus Packaging and Detection
1.4.6 The Effect of Packaging Conditions
References
Chapter 2: Assays Based on Pseudotyped Viruses
2.1 Introduction
2.2 Development of In Vitro Assays Based on Pseudotyped Viruses
2.2.1 Establishment and Optimization of In Vitro Assays Based on Pseudotyped Viruses
2.2.1.1 Selection of Target Cells
2.2.1.2 Optimization of Cell Inoculation
2.2.1.3 Optimization of the Amount of Pseudotyped Virus
2.2.1.4 Optimization of Infection Conditions
2.2.1.5 Optimization of Culture Time
2.2.1.6 Validation of In Vitro Assays Based on Pseudotyped Viruses
2.2.1.7 Specificity
2.2.1.8 Accuracy
2.2.1.9 Linear Range
2.2.1.10 Precision
2.3 Development of In Vivo Assays Based on Pseudotyped Viruses
2.3.1 Selection of Model Animals
2.3.2 Optimization of Infection Pathway
2.3.3 Determination of Infection Dose of Pseudotyped Virus
2.3.4 Comparison of Pseudotyped and Live Virus Infection Model
2.4 Conclusion
References
Chapter 3: Application of Pseudotyped Viruses
3.1 Analysis of Viral Infectivity
3.1.1 Receptor Usage
3.1.2 Cellular Tropism
3.2 In Vitro Pseudovirion-Based Neutralization Assay (PBNA)
3.2.1 Development and Evaluation of Vaccines
3.2.2 Screening and Validation of Monoclonal Neutralizing Antibodies
3.3 Screening and Validation of Viral Entry Inhibitors
3.4 Animal Model of Pseudotyped Virus Infection In Vivo
3.5 Analysis of Variations in Viral Infectivity and Antigenicity
3.5.1 Viral Variants
3.5.2 Variations in Viral Glycosylation
3.6 Antibody-Dependent Cell-Mediated Cytotoxicity (ADCC)
3.7 Conclusion
References
Chapter 4: Pseudotyped Viruses for Retroviruses
4.1 Introduction to Retroviruses
4.2 The Retrovirus Genome and Replication Cycle: Important Regions for Retroviral Vectors
4.2.1 Genome Structure
4.2.2 Retroviral Proteins
4.2.3 Viral RNA Replication
4.3 Retroviral Pseudotyping Systems
4.3.1 Overview of Retroviral Pseudotyping Plasmid Systems
4.3.2 Commonly Used Retrovirus-Derived Transgene Vectors
4.3.2.1 MLV
4.3.2.2 HIV-1 and SIV
4.3.2.3 PFV
4.4 Applications of Pseudotyped Retroviruses
4.4.1 Functions of Viral Glycoproteins and Cell Entry
4.4.2 Identification and Characterization of Host Restriction
4.4.3 Discovery of Antivirals and Characterization of Drug Resistance
4.4.4 CRISPR-Cas9 Delivery and Gene Editing
4.4.5 Gene Therapy
4.4.5.1 Monogenic Blood Disorders
4.4.5.2 Cancer Immunotherapy
4.4.6 Insertional Mutagenesis: Lessons Learned
4.5 Conclusions
References
Chapter 5: Pseudotyped Virus for Papillomavirus
5.1 Introduction
5.2 Construction of Pseudotyped Papillomavirus
5.2.1 Pseudotyped Virus Packaging System Based on Virus Vector
5.2.2 Pseudotyped Virus Packaging System Based on Plasmid Transfection
5.3 Application of Pseudotyped Papillomavirus
5.3.1 The In Vitro L1 Pseudotyped Virus Based Neutralization Assay (PBNA)
5.3.1.1 PBNA Based on Green Fluorescent Protein (GFP) and Secretory Alkaline Phosphatase (SEAP)
5.3.1.2 PBNA Based on Secretory Membrane-Anchored Luciferase (Gaussia Luciferase, Gluc)
5.3.1.3 Multiple-Color PBNA Based on Fluorescent Protein
5.3.2 In Vitro L2 Pseudotyped Virus Neutralization Test
5.3.2.1 L2-Based PBNA Mimicking In Vivo Infection
5.3.2.2 Detection of L2 Pseudotyped Virus Neutralizing Antibody Based on Furin Cleavage Intermediates
5.3.3 Animal Model of HPV Pseudotyped Virus Infection
5.3.3.1 Mouse Model of HPV Pseudotyped Virus Infection
5.3.3.2 Chimeric Pseudotyped Virus Animal Model
5.3.4 Other Applications of Pseudotyped Papillomavirus
5.4 Conclusion
References
Chapter 6: Pseudotyped Viruses for Marburgvirus and Ebolavirus
6.1 Introduction
6.2 The Biological Characteristics of Marburgvirus and Ebolavirus
6.2.1 Morphology and Genome Structure
6.2.2 Virus Entry
6.3 Drugs and Vaccines for Marburgvirus and Ebolavirus
6.4 Construction of Pseudotyped Marburgvirus and Ebolavirus
6.4.1 VSV-Based Pseudotyped Marburgvirus and Ebolavirus
6.4.2 Lentiviral-Based Pseudotyped Marburgvirus and Ebolavirus
6.4.3 Influenza-Based Pseudotyped Marburgvirus and Ebolavirus
6.4.4 Comparison of Different Types of Pseudotyped Filoviruses and Authentic Filoviruses
6.5 Application of the Pseudotyped Viruses
6.5.1 Neutralization Antibody Detection
6.5.1.1 Vaccine Efficacy Evaluation
6.5.1.2 Therapeutic Antibody Analysis
6.5.1.3 Investigation of the Kinetics of Convalescent Sera
6.5.2 Antibody-Dependent Enhancement (ADE) Evaluation
6.5.3 Study of the Mechanisms of Filovirus Infection
6.5.3.1 Mapping the Key Domains and Amino Acids on the GP
6.5.3.2 Cell Tropism Examination
6.5.3.3 Proteolytic Enzyme Analysis
6.5.3.4 Discovery and Analysis of Receptor NPC1
6.5.3.5 Receptor Tyrosine Kinase (RTK)-Related Studies
6.5.3.6 Glycosylation and Acylation Analysis
6.5.3.7 Studies of Other Host Factors
6.5.3.8 Comparisons Between Filoviruses
6.5.4 Virus Entry Inhibitor Screening
6.5.4.1 In Vitro Screening
6.5.4.2 In Vivo Verification
6.5.5 Analysis of Mutations Within the GP
6.6 Summary
References
Chapter 7: Pseudotyped Viruses for Coronaviruses
7.1 Biological Characteristics of Coronavirus
7.1.1 Structure of Coronaviruses
7.1.2 Infection of Coronaviruses
7.1.3 Diversity for Each Coronavirus
7.2 Construction of Pseudotyped Viruses for Coronaviruses
7.2.1 CoV Pseudotyped Virus Based on Vesicular Stomatitis Virus (VSV)
7.2.2 CoV Pseudotyped Virus Based on Human Immunodeficiency Virus (HIV)
7.2.3 Construction of CoV Pseudotyped Virus Based on Other Packaging Systems
7.2.4 Construction of CoV Pseudotyped Virus Based on Protein-Autonomous Packaging Systems for Virus like Particles
7.3 Application of the HCoV Pseudotyped Viruses
7.3.1 Infectivity of Highly Pathogenic hCoV and the Possibility of Cross-Species Transmission
7.3.2 Study of Highly Pathogenic hCoV Mutant Strains
7.3.3 Pseudotyped Virus for Quantifying hCoV Neutralizing Antibodies
7.3.3.1 Natural Infection
7.3.3.2 Pseudotyped Virus for CoV Vaccine Development and Clinical Evaluation
7.4 Summary
References
Chapter 8: Pseudotyped Viruses for Influenza
8.1 Introduction
8.2 Production of Pseudotyped Viruses and Developing Assays Based on Pseudotyped Viruses
8.2.1 Materials
8.2.2 Protocols for Production of Pseudotyped Viruses and Developing Assays
8.2.2.1 Production of Influenza Hemagglutinin (HA) and Neuraminidase (NA) Pseudotypes (PV)
8.2.2.2 Titration of Influenza Hemagglutinin (HA) PV
8.2.2.3 Titration of H11_Neuraminidase (H11_NA) PV via Enzyme Linked Lectin Assay (pELLA)
8.2.2.4 Pseudotype Microneutralization (pMN) Assay Using HA PV (Fig. 8.5)
8.2.2.5 Inhibition of H11_NA(X) PV by Antisera and Monoclonal Antibodies via Enzyme-Linked Lectin Assay (pELLA) (Fig. 8.6)
8.2.2.6 AutoPlate Analysis
8.3 Commentary
8.3.1 Background
8.3.2 Critical Parameters and Troubleshooting
8.3.3 Understanding Results
8.3.4 Time Considerations
References
Chapter 9: Pseudotyped Virus for Henipavirus
9.1 General Information About Henipaviruses
9.1.1 Transmission of HNV Viruses
9.1.2 Structure of HNV
9.1.3 Diversity of HNV
9.2 Construction of Pseudotyped Viruses
9.2.1 Pseudotyped Viruses Using the Moloney Murine Leukemia Virus (MuLV) Packaging System
9.2.2 Pseudotyped Viruses Using the Human Immunodeficiency Virus Type 1 (HIV-1) Packaging System
9.2.3 Pseudotyped Viruses Using the Vesicular Stomatitis Virus (VSV) Packaging System
9.2.4 Pseudotyped Viruses Using the VSV-NiV-SEAP Novel Packaging System
9.2.5 Pseudotyped Viruses Using the Self-Assembling NiV-M-VLP Packaging System
9.3 Applications of HNV Pseudotyped Viruses
9.3.1 Studies of Potential Virus Receptors
9.3.2 Viral Infectivity Studies
9.3.3 Evaluation of Neutralization Detection Systems and Potential Antibody Candidates
9.3.4 Screening Studies of Inhibitory Drugs
9.4 Conclusions
References
Chapter 10: Pseudotyped Viruses for Lyssavirus
10.1 General Information About Lyssavirus
10.2 General Information About RABV
10.2.1 Glycoprotein Structure and Its Biological Role
10.2.2 Variation and Mutation on RABV G Protein
10.3 Construction of RABV G Protein Pseudotyped Virus
10.4 Application of Pseudotyped RABV
10.4.1 Application of Pseudotyped RABV in Neutralizing Antibody Detection
10.4.2 Application of Pseudotyped RABV in mAb Screening and Epitope Mapping
10.4.3 Application of Pseudotyped RABV in Evaluation of Viral Infection, Cell Tropism, and Antigenicity
10.4.4 Application of Pseudotyped RABV in Screening of Antiviral Drugs
10.5 Summary
References
Chapter 11: Pseudotyped Viruses for Enterovirus
11.1 Introduction of Enterovirus
11.2 Biological Characteristics of Enterovirus
11.2.1 Classification of Enterovirus
11.2.2 Genome and Life Cycle of the Enterovirus
11.3 Construction of Pseudotyped Enterovirus
11.3.1 Plasmid Construction
11.3.2 Preparation of Pseudotyped Enterovirus
11.3.3 Pseudotyped Enterovirus Shows High Physical, Chemical, and Antigenic Similarities with Wild-Type Virus
11.3.4 Investigation of Encapsidation Efficiency of Pseudotyped Enterovirus
11.3.4.1 Sequence Accuracy Is the Key to the Successful Package of Pseudotyped Enterovirus
11.3.4.2 Compatibility of Trans-packaging
11.4 The Application of Pseudotyped Enterovirus
11.4.1 An Useful Tool of Studying Molecular Virology of Enterovirus
11.4.2 An Useful Tool for Detection of NtAb
11.4.2.1 The Advantage of the NtAb Detection Based on Pseudotyped Enterovirus
11.4.2.2 The Principle of pvNA
11.4.2.3 Clinical Application of pvNA for Measurement of NtAb in Human Samples
11.4.3 A Safe, Sensitive, and Visualizing Model with Pseudotyped Enterovirus
11.4.4 Screening and Evaluation of Anti-enterovirus Drugs
11.5 Summary
References
Chapter 12: Pseudotyped Viruses for Orthohantavirus
12.1 Introduction
12.2 General Property of Orthohantavirus
12.2.1 Orthohantavirus Particle and Genome
12.2.2 Orthohantavirus Entry Pathway
12.2.3 Structure and Function of Orthohantavirus Glycoproteins
12.3 Construction of Pseudotyped Orthohantaviruses
12.3.1 Construction of Replication-Deficient Pseudotyped Orthohantaviruses
12.3.1.1 The VSV-Based Packaging System
12.3.1.2 The LV-Based Packaging System
12.3.1.3 The MLV-Based Packaging System
12.3.2 Construction of Replication-Competent Pseudotyped Orthohantaviruses
12.4 Applications of Pseudotyped Orthohantaviruses
12.4.1 Mechanistic Study for Viral Entry and Infection
12.4.1.1 Identification of Cellular Receptors and Factors
12.4.1.2 Identification of Key Amino Acid in Gn/Gc
12.4.1.3 Cell Tropism
12.4.2 Quantification of Neutralizing Antibodies
12.4.3 Antigenic Property Study
12.4.4 Identification of Viral Entry Inhibitors
12.4.5 Vaccine Approach
12.5 Conclusions
References
Chapter 13: Pseudotyped Viruses for Phlebovirus
13.1 Biological Characteristics of Phlebovirus
13.1.1 Structure of Rift Valley Fever Virus (RVFV)
13.1.2 Molecular Evolution
13.2 Construction of Pseudotyped RVFV
13.2.1 Construction of Pseudotyped RVFV Using Lentiviral Vectors
13.2.2 Construction of Pseudotyped RVFV Using VSV-Based Vectors
13.2.3 Construction of Pseudotyped RVFV Using the Self-Assembly System
13.3 Application of Pseudotyped RVFV
13.3.1 Neutralizing Assay Based on Pseudotyped RVFV
13.3.2 Visual In Vivo Neutralizing Antibody Evaluation Model
13.3.3 The Mechanism of Viral Infection
13.3.4 Pseudotyped RVFV as a Candidate Vaccine
13.3.5 Neutralization Sensitivity Analysis of Natural and Artificial RVFV Variants
13.4 Conclusion
References
Chapter 14: Pseudotyped Virus for Bandavirus
14.1 Introduction
14.2 General Property of Dabie Bandavirus
14.2.1 Virion Structure and Genome Characteristics
14.2.2 Infection Mechanisms of Dabie Bandavirus
14.2.3 Genetic Diversity
14.2.4 Characteristics of the Glycoprotein
14.3 Construction of Pseudotyped Dabie Bandavirus
14.3.1 VSV-Based System
14.3.2 Lentiviral-Based System
14.4 Application of the Pseudotyped Dabie Bandavirus
14.4.1 Pseudotyped Dabie Bandavirus as Vaccine
14.4.2 Analysis of Neutralizing Antibody
14.4.3 Analysis of Viral Tropism and Entry
14.4.4 Infectivity and Neutralization Analysis of Pseudotyped Dabie Bandavirus Mutants
14.5 General Conclusions
References
Chapter 15: Pseudotyped Viruses for Mammarenavirus
15.1 Introduction
15.2 Biological Characteristics of Mammarenavirus
15.2.1 Morphology and Genome Structure
15.2.2 Replication of the Viral Genome
15.2.3 Pathogenicity
15.2.4 Diversity of LASV and LCMV
15.3 Construction of Pseudotyped Mammarenaviruses
15.3.1 The Lentiviral Vectors
15.3.2 VSV-Based Vector
15.3.3 MLV-Based Vectors
15.3.4 Recombinant Mammarenaviruses
15.3.5 The Genus of Successful Constructed Pseudotyped Mammarenaviruses
15.4 Application of the Pseudotyped Mammarenaviruses
15.4.1 The Analysis of Virus-Receptor Interactions and Host Range
15.4.2 The Mechanism of Viral Infection, Endocytosis, and Fusion
15.4.3 Detection of Neutralizing Antibodies and Evaluation of Candidate Vaccines
15.4.4 High-Throughput Screening of Viral Inhibitors
15.4.5 Analysis on the Virulence Mechanisms of Viral Mutants
15.4.6 Functional Analysis of N-linked Glycans of GPC
15.5 Conclusion
References
Chapter 16: Pseudotyped Viruses for the Alphavirus Chikungunya Virus
16.1 Biological Characteristics of Chikungunya Virus
16.1.1 Molecular Structure
16.1.2 Genotypes and Variants
16.1.3 Pathogenic Mechanisms and Biosafety Risk
16.2 Construction of Pseudotyped CHIKV
16.2.1 Construction of Pseudotyped CHIKV Using Different Vectors
16.2.1.1 Lentiviral Vectors
16.2.1.2 VSV-Based Vectors
16.2.1.3 MLV-Based Vectors
16.2.2 CHIKV Infectious Clones and Virus-like Particles
16.3 Application of Pseudotyped CHIKV
16.3.1 Neutralizing Assay Based on Pseudotyped CHIKV
16.3.1.1 Correlation of PBNA and PRNT
16.3.1.2 Factors Closely Related to CHIKV PBNA
16.3.2 Establishment of an in Vivo Imaging Model of Small Animals
16.3.3 Use of a Capture Antigen in CHIKV IgM Detection
16.3.4 Drug Screening
16.3.5 The Mechanism of Viral Infection
16.4 Conclusion
References
Chapter 17: Pseudotyped Virus for Flaviviridae
17.1 Introduction
17.2 Construction Strategies of Pseudotyped Flaviviridae
17.2.1 Construction Strategies of HCV Pseudotyped Virus
17.2.2 Construction Strategies of JEV Pseudotyped Virus
17.2.3 Construction Strategies of DENV Pseudotyped Virus
17.2.4 Construction Strategies of ZIKV Pseudotyped Virus
17.3 Application of Pseudotyped Flaviviridae
17.3.1 Study the Interaction between Virus and Host Cell
17.3.2 Neutralizing Antibody Detection and Vaccine Effect Evaluation
17.3.3 Screening of Antiviral Drugs
17.3.4 Research on Antitumor Therapy
17.4 Summary and Prospect
References
Chapter 18: Replicating-Competent VSV-Vectored Pseudotyped Viruses
18.1 Construction of Replicating-Competent VSV Viruses
18.1.1 History of Replicating-Competent VSV Viruses
18.1.2 Rescue Method of Replicating-Competent VSV Viruses
18.2 Application of Replicating-Competent VSV
18.2.1 Screening of Viral Host Factors/Receptors
18.2.2 Screening of Mutations that Escape Therapeutic mAbs
18.2.3 Vaccine Development
18.2.3.1 VSV-Based EBOV Vaccine
18.2.3.2 VSV-Based SARS-CoV-2 Vaccine
References