Basics of Chimeric Antigen Receptor (CAR) Immunotherapy

Basics of Chimeric Antigen Receptor (CAR) Immunotherapy
Copyright
Preface
	References
1. An Introduction to CAR Immunotherapy
	Immunotherapy: Historical Perspective
	Concepts That Led to the Development of CAR-Based Immunotherapy Against Cancers
		Immune Escape Through Disruption of Antigen Presentation and Costimulation
		Immune Escape Through T-Cell Exhaustion
		Tolerance as a Major Immune Escape Mechanism
		Loss of Tumor Immunogenicity
		Immune Tolerance: A Natural Checkpoint Protecting Cancers From Immune Recognition and Eradication
	References
2. Components and Design of Chimeric Antigen Receptors
	Prototypical Design of Chimeric Antigen Receptors
		CAR Ectodomain
		Significance of Choosing Antibody Domains (CARbodies) for the CARs
		Choice of Hinges Supporting scFvs
		Choice of CAR Transmembrane Domains
		Choice of CAR Endodomains and Costimulatory Molecules
	References
3. scFv Cloning, Vectors, and CAR-T Production in Laboratory for Preclinical Applications
	Advantages of Using scFv Over Full-Length Monoclonal Antibody in CAR Therapy
	Choices of Tumor Antigens, the Molecular Biology Techniques Used to Generate and Test Tumor Antigen-Specific scFvs
		scFv Generation Using Hybridoma
		scFv Screening Through Phage Display Technology
		Generating Chimeric Gene Sequences for Cloning Into Retroviral Vectors
		Retroviral Vectors Used for Cloning CAR Chimeric Genes
		Retroviral Packaging System for CAR Production
		Transposon-Based System for CAR Cloning and Expression
	Vectors Used for CAR Production
	Protocol for CAR Production in Laboratory Settings for Preclinical Applications
		Production of Third-Generation CAR Lentiviral Particles
			Materials
			Protocol 1
			Protocol 2
			Protocol 3
			Day 0
			Day 2
			Day 3
		Expansion of Primary T Cells and Generation of CAR-Modified T cells
		In Vitro Testing of CAR-T Cells
			Measuring tumor-killing activity
		In Vivo Testing of CAR-T Cells
	References
4. Production of CAR-T Cells for Clinical Applications
	Clinical Production of CAR-T Cells Using Good Manufacturing Practices
	Patient Recruitment and Eligibility Criteria for Receiving CAR T-Cell Therapy
		Eligibility Criteria for CD19+ B-Cell Malignancies
		Eligibility Criteria for CAR-T Therapy in Patients With Solid Cancers, e.g., TAG72+ Metastatic Colorectal Cancer, and FR+ O ...
	Leukapheresis and Patient T-Cell Isolation
		The Leukapheresis
		Activation and Expansion of PBMCs
	Generation of CAR-T Cells
		Lentiviral Transduction
			Lentiviral transduction in CD3/CD28 bead-activated T cells
			Retroviral transduction in OKT3-activated T cells
	Infusion of CAR-T Cells
		Patient Monitoring After T-Cell Infusion
			Adverse outcome
			Response evaluation in solid cancers
			Tumor burden in B-cell malignancies and response evaluations
			Other techniques used to assess tumor responses to CAR-T therapy
	References
5. Challenges and Opportunities to Improve CAR T-Cell Therapy
	CAR T-Cell Exhaustion
	Controlling Antigen Escape in CAR-T Cells
	Monitoring CAR-T Persistence In Vivo
	Toxicities Associated With CAR Therapy and Management of Those Toxicities
		The Cytokine Release Syndrome
		On-Target/Off-Tumor Toxicity
	Allogeneic CARs
	References
Index
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