Vaccine research and development is advancing at an unprecedented pace, with an increasing emphasis on rational design based upon a fundamental und- standing of the underlying molecular mechanisms. The aim of this volume is to provide a selection of contemporary protocols that will be useful to both novice and advanced practitioner alike. The variety of procedures required to design, develop, produce, and assess a vaccine is immense and covers aspects of ch- istry, biochemistry, molecular biology, cell biology, and immunology. No single volume can hope to cover these topics exclusively. Rather, here we attempt to provide a methods sourcebook focusing on hands-on practical advice. Comp- mentary and background information may be found in other volumes in the Me- ods in Molecular Medicine series. Of particular interest are volumes on Dendritic Cell Protocols, Interleukin Protocols, Vaccine Adjuvants, and DNA Vaccines. Since the publication of the first edition of Vaccine Protocols there have been major advances, particularly in the areas of bacterial genomics, antig- specific T-cell quantification, genetic manipulation of vaccine vectors, the h- nessing of natural molecules concerned with the regulation of immune responses, and the burgeoning field of DNA vaccinology. Hence, the extensive revision of this edition with new chapters on live viral vaccine vectors, atte- ated bacterial vectors, immunomodulators, MHC-peptide tetrameric complexes, and the identification of vaccine candidates by genomic analysis. Additionally, chapters from the first edition have been updated to accommodate state-of-t- art methods in vaccinology.

Overview of Vaccines Gordon Ada Temperature-Sensitive Mutant Vaccines Craig R. Pringle Live Viral Vectors: Construction of a Replication-Deficient Recombinant Adenovirus Anthony R. Fooks Live Viral Vectors: Vaccinia Virus Caroline Staib and Gerd Sutter Live Viral Vectors: Semliki Forest Virus Gunilla B. Karlsson and Peter J. Liljeström Development of Attenuated Salmonella Strains That Express Heterologous Antigens Frances Bowe, Derek J. Pickard, Richard J. Anderson, Patricia Londono-Arcila, and Gordon Dougan Expression and Delivery of Heterologous Antigens Using Lactic Acid Bacteria Mark A. Reuter, Sean Hanniffy, and Jerry M. Wells Synthetic Peptides Michael J. Francis Genetic Detoxification of Bacterial Toxins Mariagrazia Pizza, Maria Rita Fontana, Vincenzo Scarlato, and Rino Rappuoli Preparation of Polysaccharide-Conjugate Vaccines Carla C. A. M. Peeters, Patrick R. Lagerman, Odo de Weers, Lukas A. Oomen, Peter Hoogerhout, Michel Beurret, Karen M. Reddin, and Jan T. Poolman Adjuvant Formulations for Experimental Vaccines Duncan E. S. Stewart-Tull Incorporation of Immunomodulators into Plasmid DNA Vaccines Linda S. Klavinskis, Philip Hobson, and Andrew Woods Microencapsulation of Vaccine Antigens David H. Jones Lyophilization of Vaccines: Current Trends Gerald D. J. Adams Stimulation of Mucosal Immunity David J. M. Lewis and Christopher M. M. Hayward Induction and Detection of T-Cell Responses Kingston H. G. Mills Construction of MHC Class I-Peptide Tetrameric Complexes for Analysis of T-Cell-Mediated Immune Responses Rachel V. Samuel and Tomas Hanke Assessment of Functional Antibody Responses Ray Borrow and Paul Balmer The Use of Complete Genome Sequences in Vaccine Design Nigel J. Saunders and Sarah Butcher Severe Combined Immunodeficient (SCID) Mice in Vaccine Assessment Michael J. Dennis Clinical Trials Paddy Farrington and Elizabeth Miller Assuring the Quality and Safety of Vaccines: Regulatory Expectations for Licensing and Batch Release Elwyn Griffiths and Ivana Knezevic DNA Vaccination: An Update Douglas B. Lowrie From Vaccine Research to Manufacture: A Guide for the Researcher Nigel Allison and Howard S. Tranter Index

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