Vaccine Design - GBV · 6. Immunopotentiating Reconstituted Influenza Virosomes (IRIV) 331 7. Examples of IRIV-Based Vaccines 335 8. Preclinical Evaluations of IRIV-Based Vaccines

Vaccine DesignThe Subunit and AdjuvantApproach

Edited by

Michael F. PowellGenentech, Inc.South San Francisco, California

and

Mark J. NewmanVaxcel, Inc.Norcross, Georgia

Assistant EditorJessica R. BurdmanGenentech, Inc.South San Francisco, California

With a Foreword by Jonas Salk

Plenum Press • New York and London

Contents

List of Abbreviations xxxix

Chapter 1Immunological and Formulation Design Considerations for Subunit VaccinesMark J. Newman and Michael F. Powell

1. Introduction 12. Characterizations of Immune Responses Relevant to Vaccine Design 2

2.1. Adaptive and Innate Immunity 32.2. Antigen Processing and Presentation to the Immune System 32.3. Cytokines 62.4. Immunological Memory 7

3. Immune Responses to Infectious Pathogens, Tumors, and Vaccines 83.1. Immune Responses against Extracellular Bacteria 83.2. Immune Responses against Intracellular Bacteria 103.3. Immune Responses to Viruses 123.4. Immune Responses against Parasites 143.5. Nontraditional Uses of Vaccine Technologies 16

4. Design Criteria for Subunit Vaccines 174.1. Selection of Vaccine Immunogen 174.2. The Use of Adjuvants to Alter Vaccine Immunogenicity 184.3. Immunization/Vaccination Schedule 214.4. Vaccine Delivery Route 234.5. Vaccine Formulation Stability 234.6. Adjuvant Formulation Concerns 26

5. Conclusions 27References 28

Chapter 2Public Health Implications of Emerging Vaccine TechnologiesDale N. Lawrence, Karen L Goldenthal, John XV. Boslego,Donna K. F. Chandler, and John R. La Montagne1. Introduction 43

1.1. The Children's Vaccine Initiative 441.2. Unconquered Pathogens and Other Clinical Indications 441.3. Historical Model of Disease Eradication: Smallpox 45

xvii

xviii Contents

1.4. Recent Model for Disease Control Approaching Eradication:Haemophilus influence Type b 45

2. Reevaluation of the Premises Underlying Public Health Vaccine Practices . . . . 462.1. Premises Underlying Public Health Vaccine Programs and Practices 462.2. New Technologies Altering the Public Health Vaccine Premises 47

3. Public Health Impact of New Technologies 513.1. Research and Development 513.2. Government Agency Activities 523.3. Private and Other Manufacturers of Vaccines and

Possible International Partnerships 563.4. Opportunities for Scientific and Economic Advancement 56

4. Summary 57References 57

Chapter 3Preclinical Safety Assessment Considerations in Vaccine DevelopmentJeanine L. Bussiere, George C. McCormick, and James D. Green

1. Introduction 612. Potential Adverse Effects Associated with Vaccines 62

2.1. General Toxicity of Vaccines 622.2. Enhancement of Disease/Infection 632.3. Antigenic Competition 642.4. Unwanted Immune Responses 652.5. Cross-Reactive Antibodies 652.6. Teratogenic and Reproductive Effects 67

3. Risks Experienced with Specific Types of Vaccines and Adjuvants 673.1. Live Vaccines 673.2. Killed Vaccines 683.3. Live Carrier Vaccines 693.4. Genetically Engineered Vaccines 703.5. Adjuvants 71

4. Balancing Risks and Benefits 725. Preclinical Safety Testing Considerations 73

References 75

Chapter 4Regulatory Considerations in Vaccine DesignLawrence W. Davenport1. Introduction 812. Vaccine Development and Regulatory Issues 833. Center for Biologies Evaluation and Research 894. Code of Federal Regulations 895. Investigational New Drug Application 906. Product License and Establishment License Applications 92

Contents x j x

7. Guideline and Points to Consider Documents 948. Summary 95

References 95

Chapter 5Clinical Considerations in Vaccine Trials with Special Reference to Candidate HIVVaccinesPatricia E. Fast, Leigh A. Sawyer, and Susan L. Wescott1. Introduction 972. Vaccine Candidates Currently in Clinical Trials 98

2.1. Vaccine Candidates Other Than HIV-1 982.2. Types of HIV-1 Vaccine Trials: Preventive, Therapeutic, and Perinatal . . 1032.3. HIV-1 Vaccine Candidates 1032.4. Sequential Use of Different Vaccines in One Regimen 104

3. Clinical Studies of Vaccine Candidates 1073.1. Approach to Vaccine Trials 1073.2. Phases of Clinical Development 107

4. Factors in Design of Clinical Trials 1104.1. Ethical Considerations in Testing HIV Vaccines 1104.2. Volunteer Selection Ill4.3. Special Populations 1124.4. Informed Consent 1134.5. Age of Volunteers for HIV-1 Vaccine Trials 1144.6. Choice of Dose or Dose Range for Clinical Trials 1144.7. Risks of Vaccination 1154.8. Reimbursement 1174.9. Motivation of Volunteers in HIV-1 Vaccine Trials 1184.10. Randomization, Placebos, and Blinding 1184.11. Special Considerations for Vaccine Trials in Pregnant Women 1194.12. Special Considerations for Trials in Developing Countries 120

5. Trial Outcome Measures 1215.1. Laboratory Assays in HIV-1 Prophylactic Trials: The Dilemma of

Unknown Correlate(s) of Immunity/Protection 1215.2. Outcome Measures in HIV-1 Therapeutic Trials: The Dilemma of

Unknown Surrogate Marker(s) 1226. Conclusions 123

References 124

Chapter 6Laboratory Empiricism, Clinical Design, and Social Value: The Rough Roadtoward Vaccine DevelopmentDonald P. Francis1. Introduction 1-352. Discussion 135

xx Contents

3. Conclusion 139References 139

Chapter 7A Compendium of Vaccine Adjuvants and ExcipientsFrederick R. Vogel and Michael F. Powell

Introduction 141Adju-Phos 142AF (see Antigen Formulation)Algal Glucan 143Algammulin 145Alhydrogel 146Alum (see Alhydrogel, Rehydragel)Aluminum Phosphate Gel (see Adju-Phos)Arlacel 85 (see Span 85)Antigen Formulation 147Avridine® 148BAYR1005 149Block Copolymer (see CRL 1005)Calcitriol I 5 0

Calcium Phosphate Gel 151CFA (see Freund's Complete Adjuvant)Cholera Holotoxin (CT) 152Cholera Toxin B Subunit (CTB) 152CRL1005 154Cytokine-Containing Liposomes 156DDA 157Deoxycholic Acid (see DOC/Alum Complex)DHEA 158Dihydroxy vitamin D3 (see Calcitriol)Dimethylammonium Bromide (see DDA)DMPC 159DMPG 160DOC/Alum Complex 161Freund's Complete Adjuvant 162Freund's Incomplete Adjuvant 163Gamma Interferon (see Interferon-}')Gamma Inulin 164Gerbu Adjuvant 165GM-CSF 166GMDP 167IFA (see Freund's Incomplete Adjuvant)IL-ly? Peptide (see Sclavo Peptide)Imiquimod 168ImmTher™ 169

Contents xxi

Immune Stimulating Complex (see ISCOM)Interferon-}' 170Interleukin-ip 171Interleukin-2 172Interleukin-7 174Interleukin-12 175ISCOM(s)™ 177Isoprep 7.0.3™ 178Liposomes 179Loxoribine 181LT-OA or LT Oral Adjuvant 182MF59 183Monophosphoryl Lipid A (see MPL®)Montanide®ISA51 184Montanide® ISA 720 185MPL® 186MTP-PE 188MTP-PE Liposomes 189Murametide 190Muramyl Dipeptide (see Threonyl-MDP)Murapalmitine 191D-Murapalmitine 192NAGO 193Nonionic Surfactant Vesicles 195Phosphatidylcholine (see DMPC)Phosphatidylglycerine (see DMPG)Pleuran '96PLGA, PGA, and PLA 1 9 8

PluronicL121 2 0 °PMMA 2 0 1

PODDS™ 2 0 2

Poloxamer401 (seePluronic L121)Polymethyl Methacrylate (see PMMA)Poly rA: Poly rU 2 0 3

Polyphosphazene 2 ^Polysorbate 80 2 0 5

Protein Cochleates 2 0 6

Proteinoid Microspheres (see PODDS™)QS-21 2 0 8

QuilA 2 I ( )

Rehydragel HPA 2U

Rehydragel LV 2 1 2

S-28463 2 1 3

SAF-1 2 1 4

Saponin (see Iscoprep, Quil A, QS-21)

xxii Contents

Sclavo Peptide 215Sendai Proteoliposomes 216Sendai-Containing Lipid Matrices 216Sorbitan Trioleate (see Span 85)Span 85 218Specol 219SPT (see Antigen Formulation)Squalane 220Squalene 221Stearyl Tyrosine 222Stimulon™ (see QS-2)Termurtide™ (see Threonyl-MDP)Theramide™ 223Threonyl-MDP 224Tween 80 (see Polysorbate 80)Ty Particles 225Walter Reed Liposomes 226Acknowledgments and Update Summary 227

Chapter 8Adjuvant Properties of Aluminum and Calcium CompoundsRajesh K. Gupta, Bradford E. Rost, Edgar Relyveld, and George R. Siber1. Introduction 2292. Aluminum Compounds 230

2.1. Method of Preparation 2302.2. Factors Affecting the Adsorption 2312.3. Adjuvant Properties 2332.4. Mechanism of Action 2372.5. Limitations 238

3. Calcium Phosphate 2393.1. Method of Preparation 2393.2. Adjuvant Properties 241

4. Combination of Aluminum Compounds with Other Adjuvants 2415. Summary 242

References 242

Chapter 9Structure and Properties of Aluminum-Containing AdjuvantsStanley L. Hem and Joe L. White1. Introduction 2492. Characterization 249

2.1. Aluminum Hydroxide Adjuvants 2512.2. Aluminum Hydroxyphosphate Adjuvants 2532.3. Commercial Vaccines 257

3. Optimizing Antigen-Adjuvant Adsorption 258

Contents xxiii

3.1. Model Proteins 2593.2. Malaria Antigens 262

4. Effect of Protein Adsorption on Surface Properties 2655. Interactions in Multivalent Vaccines Using Mixtures of Aluminum-Containing

Adjuvants 2675.1. Redistribution of Antigens 2675.2. Redistribution of Phosphate Anions 2685.3. Colloidal Interactions 269

6. Effect of Anions 2706.1. Aluminum Hydroxide Adjuvant 2716.2. Aluminum Phosphate Adjuvant 272


Chapter 10MF59: Design and Evaluation of a Safe and Potent Adjuvant for Human VaccinesGary On, Gail L. Barchfeld, David Chernoff, Ramachandran Radhakrislman,Peter van Hoogevest, and Gary Van Nest1. Rationale for and Design of Microfluidized Oil/Water Emulsions 2772. Preclinical Experience with MF59 2813. Mechanism of Adjuvant Activity 2854. Manufacturing and Scaleup of MF59 2885. Clinical Results with the MF59 Adjuvant Formulation 2936. Summary 294

References 2 9 4

Chapter 11Development of Vaccines Based on Formulations Containing Nonionic BlockCopolymersRobert N. Brey1. Introduction2. Vaccine Delivery Systems and Adjuvants 2 "3. Nonionic Block Polymers in Vaccine Formulations 3004. Properties of Vaccine Formulations Containing Nonionic Block Copolymers . . 307

5. Summary 3 0 8

References 3 0 8

Chapter 12Development of an Emulsion-Based Muramyl Dipeptide AdjuvantFormulation for VaccinesDeborah M. Lidgate and Noelene E. Byars1. Introduction2. Compound Selection3. Chemistry of Threonyl-MDP 3 1 5

xxiv Contents

4. Emulsion Formulation Considerations 3155. SAF Formulation 3166. SAF Manufacturing 3177. Final Product 3198. Mechanism of Action 3219. Studies Utilizing SAF Emulsion with Threonyl-MDP 321


Chapter 13

Liposomal Presentation of Antigens for Human VaccinesReinhard Gliick1. Introduction 3252. History of Liposomes 3263. Liposomes as Adjuvants 3274. Approaches with Liposomal Vaccines 3275. Technologies for Liposomal Vaccines 3296. Immunopotentiating Reconstituted Influenza Virosomes (IRIV) 3317. Examples of IRIV-Based Vaccines 3358. Preclinical Evaluations of IRIV-Based Vaccines 3369. Clinical Evaluation of IRIV-Based Vaccines 336

10. Patent Protection and Product Licensing of IRIV-Designed Vaccines 33911. Prospects for the Future 34012. Summary 340

References 341

Chapter 14

Liposome Design and Vaccine DevelopmentPatricia J. Freda Pietrobon1. Introduction 3472. General Properties of Liposomes 348

3. Liposomes and Enhancement of Humoral Immune Responses 3493.1. Antibody Responses to Thymus-Independent Antigens 3493.2. Liposome Stimulation of Thymus-Independent Type 1 and

Thymus-Independent Type 2 Antibody Responses 3513.3. Liposomes That Provide T-Dependent Help to Weak (T-Independent)

Antigens 3523.4. Liposomes and Enhancement of Thymus-Dependent Antibody Responses 353

4. Liposomes and Cell-Mediated Immune Responses 3544.1. Liposomal Antigen Delivery and CTL Induction 3544.2. Liposomes and Their Effect on Antiviral and Antiparasitic Immunity . . . . 355


Contents x x v

Chapter 15

Lipid Matrix-Based Vaccines for Mucosal and Systemic Immunization

Raphael J. Mannino and Susan Gould-Fogerite1. Introduction 3632. Protein Cochleate Vaccines 364

2.1. Structure 3652.2 Formulation 3662.3. Oral Immunization and the Common Mucosal Immune System 3672.4. Influenza: Immune Response to Cochleate Vaccines 3682.5. Parainfluenza Type 1: Immune Response to Cochleate Vaccines 3722.6. Human Immunodeficiency Virus: Immune Response to Cochleate Vaccines 374

3. Fusogenic Proteoliposomes 3763.1. Introduction 3763.2. Sendai Fusogenic Proteoliposomes 377

4. Unique Covalent Peptide-Phospholipid Complexes 3794.1. Introduction 3794.2. Formulation 3804.3. Stimulation of an Immune Response to Hydrophilic B-Cell Determinants

through an Association with Putative Th-Cell Determinants 3805. Summary 382

References 384

Chapter 16

Polymer Microspheres for Vaccine Delivery

Justin Hanes, Masatoshi Chiba, and Robert Langer1. Introduction 3892. Polymer Microspheres for Vaccine Delivery 390

2.1. Parenteral Immunization 3902.2. Mucosal Immunization 398

3. Polymers for Vaccine Microencapsulation 4023.1. Lactide/Glycolide Polyesters 4023.2. Polymers with Built-in Adjuvanticity 4033.3. Gelatin 4043.4. Polyphosphazene Gels 4043.5. Microencapsulated Liposomes 405

4.TheRoleofImmunostimulants 4055. Microsphere Production 4066. Consideration of Vaccine Formulation Stability 4077. Regulatory Issues 4088. Conclusions 408

References 4°9

Contents

Chapter 17Vehicles for Oral ImmunizationNoemi Santiago, Susan Haas, and Robert A. Baughman1. Oral Delivery of Antigens 413

1.1. Introduction 4131.2. Approaches for Oral Immunization 415

2. Novel Adjuvants and Vehicles for Oral Immunization 4212.1. Surfactant-Based Vehicles 4212.2. Microorganism-Based Delivery 4222.3. Microspheres 426

3. Proteinoid-Based Oral Antigen Delivery 4263.1. Review of the Technology 4263.2. Oral Immunization with Flu Antigens 4273.3. Ovalbumin as a Model Antigen 429References 433

Chapter 18Design and Production of Single-Immunization VaccinesUsing Polylactide Polyglycolide Microsphere SystemsJeffrey L. Cleland1. Introduction 439

1.1. Rationale for Single-Immunization Vaccines 4391.2. Potential Single-Immunization Vaccine Formulations 440

2. Predevelopment Studies for Design of in Vivo Release Profile 4453. Relationship Between Polymer Properties and Vaccine Design 4474. Stability Constraints on Subunit Vaccines in Polylactide Microspheres 4495. Manufacturing Hurdles for Polylactide Microsphere-Based Vaccines 451

5.1. Aseptic Manufacturing and Terminal Sterilization 4515.2. Scaleup, Process Integration, and Solvent Handling Concerns 453

6. Regulatory and Toxicology Issues 4556.1. Residual Solvent Concerns 4566.2. Histology Studies of Polylactide Formulations 457

7. Overview of Major Issues in Design of Polylactide Vaccine Formulations . . . . 4588. Future Directions in the Development of Single-Shot Subunit Vaccines 459

References 459

Chapter 19Nanoparticles as Adjuvants for VaccinesJb'rg Kreuter1. Introduction 4632. Production of Nanoparticles 464

2.1. Polymerization in the Presence of the Immunogen 4642.2. Polymerization in the Absence of the Immunogen 4642.3. Nanoparticles Produced in an Organic Phase 465

Contents xxvii

3. Characterization of Nanoparticles 4654. Influence of Particle Size and Hydrophobicity on the

Adjuvant Effect of Nanoparticles 4665. Antibody Response and Protection Induced with Vaccines

Containing PMMA Nanoparticles 4676. Body Distribution, Elimination, and Toxicological

Reactivity of PMMA Adjuvants 4697. Summary 470

References 471

Chapter 20Water-Soluble Phosphazene Polymers for Parenteral and Mucosal VaccineDeliveryLendon G. Payne, Sharon A. Jenkins, Alexander Andrianow and Bryan E. Roberts1. Introduction 4732. Microsphere Formulation and Characterization 476

2.1. Microsphere Generation 4762.2. Characterization of Microspheres 4782.3. Antigen Release 478

3. Immunogenicity Studies 4823.1. Parenteral Immunization 4823.2. Mucosal Immunization 4903.3. The Role of Polyphosphazene in Vaccine Delivery 491


Chapter 21Monophosphoryl Lipid A as an Adjuvant: Past Experiences and New DirectionsJ. Terry Ulrich and Kent R. Myers

1. Introduction 4951.1. Historical Perspective 4951.2. Rationale for Using MPL® as an Adjuvant 496

2. Characteristics of MPL® as a Product 4972.1. Manufacturing, Chemistry, and Quality Control 4972.2. Stability of MPL®—Considerations for Formulation 501

3. Methods of Using MPL® as an Adjuvant 5023.1. Aqueous (Nonparticulate) Vehicles 5033.2. Paniculate Vehicles 503

4. Past Experiences—The Effectiveness of MPL® as an Adjuvant 5054.1. Preclinical Experience 5054.2. Clinical Experiences 5134.3. Modes of Action of MPL® as an Adjuvant 516

5. New Directions 5176. Summary and Conclusions 518

References 518

xxviii Contents

Chapter 22Structural and Immunological Characterization of the Vaccine Adjuvant QS-21Charlotte Read Kensil, Jia-Yan Wu, and Sean Soltysik1. Introduction 525

1.1. Historical Use of Quillaja Saponins as Adjuvants 5251.2. Discovery of Diversity of Quillaja Saponaria Adjuvants 5261.3. Stimulon™-QS-21 Adjuvant 526

2. Chemical and Physical Properties 5262.1. Structure 5262.2. Amphipathic Character 527

3. Manufacturing 5273.1. Manufacturing and Quality Control 5273.2. Formulation 5283.3. Stability 529

4. Adjuvant Activity 5304.1. Antibody Response 5304.2. Cell-Mediated Immune Response 5334.3. Adjuvant Effect in Other Species 533

5. Mechanism of Action 5345.1. Effect of Accessory and Effector Cell Depletion on CTL and Antibody

Response 5345.2. Cytokine Profile Induced by QS-21-Adjuvanted Vaccines 5355.3. Structure/Function Studies 537

6. Clinical Experience with QS-21 5387. Summary and Future Directions 539

References 539

Chapter 23A Novel Generation of Viral Vaccines Based on the ISCOM MatrixG. F. Rimmelzwaan and A. D. M. E. Osterhaus1. Introduction 5432. The ISCOM Structure 543

2.1. Components, Chemical and Physical Properties 5432.2. Construction of ISCOMs 545

3. Induction of B- and T-Cell Responses by ISCOMs 5453.1. Induction of B-Cell Responses 5453.2. Induction of T-Cell Responses 546

4. Viral Vaccines Based on ISCOMs: Examples and State of the Art 5474.1. Influenza Viruses 5474.2. Paramyxoviruses 5504.3. Herpesviruses 5504.4. Rhabdoviruses 5514.5. Hepadnaviruses 5514.6. Retroviruses 552

5. General Considerations 552

Contents xxjx

5.1. Toxicological Aspects 5525.2. Mode of Administration 553


Chapter 24Vaccine Adjuvants Based on Gamma InulinPeter D. Cooper1. Introduction 5592. The Nature of Effective Vaccine Adjuvants 5603. Involvement of Complement in Immune Defenses 561

3.1. Opsonic Effects of ACP Activation 5623.2. Immune Modulator Effects of CRLigation 563

4. Complement Activators as Immune Modulators 5645. Inulin as Complement Activator 565

5.1. Chemistry of Inulin 5655.2. Formation of Gamma Inulin 5665.3. Inulin and Complement 566

6. Gamma Inulin as Immune Modulator 5676.1. Gamma Inulin—The Product 5676.2. Vaccine Adjuvant Effects of Gamma Inulin 5686.3. Effects of Gamma Inulin on Leukocyte-Cytokine Interactions 570

7.Algammulin—Composite of Alum and Gamma Inulin 5707.1. Algammulin—The Product 5717.2. Vaccine Adjuvant Effects of Algammulin 5717.3. Effects of Algammulin on Leukocyte-Cytokine Interactions 573

8. Safety Profile of Gamma Inulin-Based Vaccine Adjuvants 5748.1. Gamma Inulin 5748.2. Algammulin 575

9. Comparison with Other Vaccine Adjuvants 57510. Summary and Conclusions 576

References 577

Chapter 25A New Approach to Vaccine Adjuvants: Immunopotentiation by IntracellularT-Helper-Like Signals Transmitted by LoxoribineMichael G. Goodman1. Introduction 581

1.1. Discovery of Loxoribine and Its Analogues 5811.2. Overview of Cell Types Activated and Immunobiological

Actions Evoked by Loxoribine and Its Analogues 5822. Amplification of Antibody Responses 583

2.1. Cellular Mechanism of Action 5832.2. T-Helper-Like Signals 5842.3. Animal Studies 5862.4. Immunoprophylaxis 588

xxx Contents

2.5. Effects on Human Antibody Responses 5893. Enhancement of T-Cell Responses 591

3.1. Augmentation of Cytolytic T-Cell Reactivity 5913.2. Antigen-Specific T-Cell Proliferation 591

4. Role of Cytokines 5925. Biochemical Studies 593

5.1. Transport System 5935.2. Loxoribine Binding Proteins: Binding Studies, Autoradiography 5935.3. Interaction with Purine Salvage Pathway Enzymes 5945.4. Protein Kinase C Pathway Independence 596

6. Preclinical Studies 5966.1. Advantageous Features Unique to Loxoribine 5966.2. Dose Response, Vehicle, Route of Administration 5976.3. Effects on Autoimmune-Prone Mice 5996.4. Patent Status, Synthesis 5996.5. Safety Profile and Stability 6016.6. Activity Relative to Other Immunomodulators 601

7. Clinical Trials 6037.1. Phase I Trial in Advanced Cancer Patients 6037.2. Phase I Trial in Chronic Renal Failure Patients on Hemodialysis 6037.3. Future Trials 604

8. Summary and Conclusions 605References 605

Chapter 26Stearyl Tyrosine: An Organic Equivalent of Aluminum-Based ImmunoadjuvantsChristopher Penney1. Introduction 6112. Why Replace Aluminum? 6123. Stearyl Tyrosine: Overview 612

3.1. Synthesis 6143.2. Toxicity 615

4. Adjuvanticity with Bacterial Vaccines 6155. Adjuvanticity with Viral Vaccines 6176. Other Applications 6197. Analogues of Stearyl Tyrosine 6207. Conclusions 622

References 623

Chapter 27Cytokines as Vaccine Adjuvants: Current Status and Potential ApplicationsPenny Dong, Carsten Brunn, and Rodney J. Y. Ho1. Introduction 625

1.1. The Role of Cytokines in Antigen Presentation 6261.2. The Role of Cytokines in T-Cell Growth 6271.3. The Role of Cytokines in B-Cell Growth 627

Contents xxxi

2. Application of Cytokines in Modulating Antigen Presentation 6282.1. Cytokines 6282.2. Cytokine Induction by Other Adjuvants 629

3. Application of Cytokines in Modulating T-Cell Growth to EnhanceCellular Immune Response 631

4. Application of Cytokines in Modulating B-Cell Growth to Enhance HumoralImmune Response 632

5. Cytokine Fragments as Vaccine Adjuvants 6336. Local Delivery of Cytokines and the Sequence of Administration 6347. Conclusions 638

References 639

Chapter 28Cytokines as Immunological AdjuvantsAndrew W. Heath

1. Introduction 6452. Production of Cytokines 6463. Patent Protection 6464. A Brief Summary of Cytokine Adjuvanticity 647

4.1. Interleukin-1 6474.2. Interleukin-2 6484.3. Interferon-y 649

5. Means of Improving Adjuvant Effects of Cytokines 6496. Areas of Particular Potential for Cytokine Adjuvants 650

6.1. Immunodeficiency 6506.2. Direction of the Immune Response 652


Chapter 29Cytokine-Containing Liposomes as Adjuvants for Subunit VaccinesLawrence B. Lachman, Li-Chen N. Shih, Xiao-Mei Rao, Stephen E. Ullrich,and Jeffrey L. Cleland1. Introduction 6592. Cytokines as Adjuvants 6593. Presentation of Antigen by Liposomes 6604. Dehydration-Rehydration Vesicles (DRVs) Containing Cytokines 662

4.1. Preparation of DRVs 6624.2. Trapping Capacity of DRVs 6624.3. Slow Release of the Trapped Cytokines and HIV-1 MN rgp 120 in the DRVs 6634.4. Localization of Trapped Proteins 664

5. Immunization of Mice with DRVs Containing Cytokines and HIV-1 MN rgp 120 6656. Possible Mechanisms of Action of DRVs 6677. Future Directions for Cytokine-Containing DRVs 669

References 669

xxxii Contents

Chapter 30Haemophilus influenzae Type b Conjugate VaccinesPeter J. Kniskern, Stephen Marburg, and Ronald W. Ellis1. Introduction 6732. Rational Conjugate Design, Chemistry, and Process Control 675

2.1. Outline of the Processes 6752.2. Process Design and Quantitative Aspects of Covalency 6792.3. Characterization of Starting Materials and Process Intermediates 6802.4. Separation of Unconjugated Polysaccharide 682

3. Characterization of Bulk Conjugates and Final Container Vaccines 6824. Immunogenicity and Efficacy 6875. Lessons for the Future 689

5.1. The Implications of Carrier Choice and of Size of Starting Ps 6895.2. The Importance of Rigorous Process Control and Analytical Testing of the

Process Intermediates, Final Products, and Formulated Vaccines 6906. Summary 690

References 691

Chapter 31Pneumococcal Conjugate VaccinesRonald Eby1. Introduction 695

1.1. Epidemiology of Pneumococcal Disease 6951.2. Current Licensed Pneumococcal Vaccines 6971.3. The Need for Pneumococcal Conjugate Vaccines 697

2. Pneumococcal Conjugate Vaccines 6982.1. Chemical Structures 6982.2. Preclinical Analysis 7032.3. Clinical Trial Results 706


Chapter 32Lyme Vaccine Enhancement: N-Terminal Acylation of a Protein Antigen andInclusion of a Saponin AdjuvantRichard T. Coughlin, Jianneng Ma, and Daniel E. Cox1. Background 719

1.1. Lyme Disease, Incidence, Transmission, and Symptoms 7191.2. The Biochemistry of Bacterial Lipoprotein Biosynthesis 7201.3. Comparison of Acylated and Nonacylated Forms as Immunogens 72114. Attempts at Synthetic Generation of Lipoproteins 722

2. Product Description of Borrelia Burgdorferi Lipoprotein Vaccines 7252.1. E. coli Expression and Purification 7252.2. Vaccine Safety 727

Contents • xxxiii

3. Vaccine Evaluation 7273.1. What Is the Nature of Vaccine-Induced Protection? 7273.2. Differences in Immunoprotective Antibodies Induced by Acylated versus

Nonacylated Outer Surface Proteins 7293.3. Cross Neutralization of Genetically and Geographically Divergent Borrelia 729

4. Future Directions 7304.1. Progress of Clinical Trials in High-Incidence Areas of the United States . . 7304.2 Social Behavior as a Factor in the Control of Lyme Disease 730References 731

Chapter 33Vaccine Research and Development for the Prevention of Filarial NematodeInfectionsRobert B. Grieve, Nancy Wisnewski, Glenn R. Frank, and Cynthia A. Tripp1. Introduction 7372. Evidence for Protective Immunity in Filarial Nematode Infections 738

2.1. Existence of Naturally Acquired Protective Immunity 7392.2. Evidence of Concomitant Immunity in Filariasis 7432.3. Experimental Immunization Studies 745

3. Characteristics of the Ideal Vaccine 7494. Dirofilaria immitis: A Vaccine in Animals and a Model for Human Filariases . . 7505. Identification of Candidate Antigens 751

5.1. Dirofilaria immitis Antigens Recognized by Antibodies in Immune Dog Sera 7525.2. Lymphatic Filariasis 7535.3. Onchocerciasis 7535.4. Microfilaria 754

6. Cloning, Characterization, and Expression of Candidate Antigens 7547. Summary 759

References 760

Chapter 34Retrovirus and Retrotransposon Particles as Antigen Presentation and DeliverySystemsSally E. Adams and Alan J. Kingsman1. Introduction 7692. Presentation of Immunogens as Hybrid Ty-VLPs 770

2.1. The Yeast Ty Element 7702.2. Hybrid Ty-VLPs 7722.3. Immune Responses Elicited by Hybrid VLPs 772

3. Presentation of Antigens as Virus-Derived Particles 7783.1. The Construction of Hybrid Retroviral Cores 7783.2. Expression of Authentic Glycoproteins Using VDPs 780


xxxiv Contents

Chapter 35Rationale and Approaches to Constructing Preerythrocytic Malaria VaccinesStephen L. Hoffman and John B. Sacci, Jr.

1. Irradiated Sporozoite Vaccination as a Model for Malaria Vaccine Development 7872. Preventing Sporozoite Invasion of Hepatocytes 7883. Attacking the Infected Hepatocyte 791

3.1. CD8+ T-Cell-Dependent Irradiated Sporozoite Vaccine-InducedProtective Immunity 791

3.2. Adoptive Transfer of CD8+ CTLs against the CS Protein Are Protective . . 7923.3. Immunization with CS Protein Vaccines Induces CD8+ T-Cell-Dependent

Partial Protection in Rodent Malaria Model Systems 7923.4. Induction of CTLs against the P.falciparum CS Protein in Humans . . . . 7933.5. Identification of Sporozoite Surface Protein 2 as a Target of

Vaccine-Induced CD8+Protective CTLs 7943.6. CD4+ CTLs against the CS Protein Mediate Protective Immunity 7943.7. Other Parasite Antigen Targets in Infected Hepatocytes 7953.8. Interferon-y and Other Cytokines 795

4. The Future: Inducing Multiple Immune Responses against Multiple Targets . . 796References 797

Chapter 36The MAP System: A Flexible and Unambiguous Vaccine Design of BranchedPeptidesBernardetta Nardelli and James P. Tarn1. Introduction 8032. MAP Design and Synthesis 8043. MAP Antigenicity 8064. MAP Immunogenicity 806

4.1. Production of a High-Titered Antibody Response 8074.2. Reactivity of the Elicited Antisera against the Cognate Proteins 8074.3. Correlation MAP Structure and Immunogenicity 8084.4. Enhanced Immunogenicity of MAPs Containing B- and T-Cell

Epitopes, and Protection Mediated by the Immune Sera 8084.5. Generation of Long-Term Antibody Responses 8104.6. Capacity of Overcoming MHC-Associated Nonresponsiveness 8115. Lipidated MAP Vaccines 811

6. Concluding Remarks 814References 816

Chapter 37Design of Experimental Synthetic Peptide Immunogens for Prevention of HIV-1and HTLV-I Retroviral InfectionsMary Kate Hart, Thomas J. Palker, and Barton F. Haynes1. Introduction 821

Contents xxxv

2. Synthetic Peptide Approach for HIV-1 8222.1. Peptide Synthesis and Purification 8232.2. Design of HIV-1 Th-B Hybrid Synthetic Peptides 8232.3. Induction of Neutralizing Antibody Responses by Th-B

Peptide Immunogens 8252.4. Rhesus Monkey Studies 8262.5. Peptide-Induced Anti-HIV-1 Responses in Chimpanzees 8282.6. Effect of gp41 Fusogenic Domain (F) on Immune Response 8292.7. Peptide Induction of MHC Class I Molecule-Restricted Anti-HIV-1 CTLs 831

3. Simian Immunodeficiency Virus (SIV) env Synthetic Peptides InduceSIV-Specific CD8+CTLs in Rhesus Monkeys 833

4.HTLV-I 8344.1. HTLV-I Pathogenesis and Epidemiology 8344.2. Feasibility of an HTLV-I Vaccine 8344.3. Envelope Sequence Divergence among HTLV-I Strains 8354.4. Vaccine-Induced Protection from HTLV-I Challenge 8354.5. Synthetic Peptide Approaches to HTLV-I Vaccine Development 836

5. Summary and Future Directions 838References 839

Chapter 38

Design and Testing of Peptide-Based Cytotoxic T-Cell-MediatedImmunotherapeutics to Treat Infectious Diseases and Cancer

Robert W. Chesnut, Alessandro Sette, Esteban Cells, Peggy Wentworth,Ralph T. Kubo, Jeff Alexander, Glenn Ishioka, Antonella Vttiello, and Howard M. Grey1. Background 847

1.1. The Role of Cytotoxic T Cells in the Control and Elimination ofInfectious Diseases and Cancer 847

1.2. The Processing and Presentation of Antigen to CD8+ Cytotoxic T Cells . . 8491.3. Summary of Evidence Showing That Synthetic Peptides

Can Serve as Immunogens 8502. Selection of Antigenic Peptides 851

2.1. Introduction 8512.2. Definition of MHC-Binding Motifs 8512.3. Validation of the MHC-Binding Motifs 8552.4. Screening Vaccine Candidate Peptides for MHC Binding and in Vitro and

in Vivo CTL Activation 8563. Therapeutic Applications 859

3.1. Background 8593.2. Adoptive Cellular Therapy 8593.3. In Vivo Injectable Immunotherapeutics 861

4. Summary and Perspectives 866References 867

xxxvi Contents

Chapter 39Development of Active Specific Immunotherapeutic Agents Based onCancer-Associated MucinsJohn Samuel and B. Michael Longenecker1. Introduction 8752. Immune Responses Relevant to Cancer Rejection 8763. Target Antigens for ASI 8774. Cancer-Associated Mucins as Target Antigens 8785. Immune Responses to Carbohydrate Epitopes 8806. Immune Responses to Mucin Core Peptides 8827. Pharmaceutical Formulation of ASI Agents 8858. Summary 885

References 886

Chapter 40Synthetic Peptide Vaccines for SchistosomiasisDonald A. Ham, Sandra R. Reynolds, Silas Chikunguwo, Steve Furlong, andCharles Dahl1. Background 8912. Targets of Immune Response 8923. Selection of Candidate Vaccine Antigens 893

3.1. Selection of B- and T-Cell Peptide Epitopes for Sm23 and TPI 8943.2. Vaccination of Mice with TPI and Sm23MAPs 8983.3. Immunobiology of TPI and Sm23 MAP Vaccination 900


Chapter 41Synthetic Hormone/Growth Factor Subunit Vaccine with Application toAntifertility and CancerLaura L. Snyder, David V. Woo, Pierre L. Triozzi, and Vernon C. Stevens1. Introduction 9072. Special Considerations in Vaccine Development for Fertility Control 9083. Immunological Fertility Control 909

3.1. Hormones as Target Antigens 9103.2. Human Chorionic Gonadotropin 910

4. Development of an Antifertility Vaccine Based on HumanChorionic Gonadotropin 911

4.1. Peptide Immunogen Selection 9114.2. Carrier Selection 9124.3. Coupling and Ratio of Peptide to Carrier 9124.4. Adjuvant and Vehicle Selection 913

5. Preclinical Vaccine Experience in Antifertility 9136. Clinical Vaccine Experience in Antifertility 915

Contents xxxvii

7. Application to Cancer: Immunological Control of Cancer 9168. Application to Cancer: Tumor Expression of Chorionic Gonadotropin 9189. Application to Cancer: Clinical Vaccine Experience 921

10. The Future of the hCG£CTP37-DT Vaccine 92310.1. Future Development for Antifertility 92310.2. Future Development for Cancer 924References 925

Index 931