The field of DNA vaccines has undergone explosive growth in the last few years. As usual, some historical precursors of this approach can be d- cerned in the scientific literature of the last decades. However, the present state of affairs appears to date from observations made discreetly in 1988 by Wolff, Malone, Felgner, and colleagues, which were described in a 1989 patent and published in 1990. Quite surprisingly, they showed that genes carried by pure plasmid DNA and injected in a saline solution, hence the epithet naked DNA, could be taken up and expressed by skeletal muscle cells with a low but reproducible frequency. Such a simple methodology was sure to spawn many applications. In a separate and important line of experimentation, Tang, De Vit, and Johnston announced in 1992 that it was indeed possible to obtain humoral immune responses against proteins encoded by DNA delivered to the skin by a biolistic device, which has colloquially become known as the gene gun. The year 1993 saw the publication of further improvements in the me- ods of naked DNA delivery and, above all, the first demonstrations by several groups of the induction of humoral and cytotoxic immune responses to viral antigens expressed from injected plasmid DNA. In some cases, protection against challenge with the pathogen was obtained. The latter result was - questionably the touchstone of a method of vaccination worthy of the name.

Klappentext

In DNA Vaccines: Methods and Protocols, state-of-the-art review articles by leading experts summarize how to develop and employ the highly promising new DNA vaccines, what clinical results can be expected from their use, and what is known about how they work. Key topics range from vaccine design and construction to preparation and delivery methods, including the use of classical adjuvants, "genetic adjuvants," and the immunostimulatory properties of DNA and selected oligonucleotide sequences. Several contributors provide strategic ideas on antigen engineering and describe the particularly novel applications of DNA vaccine methodology that have recently emerged. In addition, there is a discussion of dendritic cells and antigen-presenting cells, the understanding of whose actions holds great promise for modulating the immune response, and thus for treating disease. Cutting-edge and comprehensive, DNA Vaccines: Methods and Protocols provides a broad panorama of the methods and thinking from which the vaccines of tomorrow will evolve, and so constitutes an invaluable sourcebook for both experts developing new applications and newcomers who want to gain mastery of the techniques and problems involved.

Zusammenfassung
" The year 1993 saw the publication of further improvements in the me- ods of naked DNA delivery and, above all, the first demonstrations by several groups of the induction of humoral and cytotoxic immune responses to viral antigens expressed from injected plasmid DNA.

Inhalt
Purification of Supercoiled Plasmid.- Production of Plasmid DNA in Industrial Quantities According to cGMP Guidelines.- Development and Characterization of Lyophilized DNA Vaccine Formulations.- Repeated Use of Qiagen Columns in Large-Scale Preparation of Plasmid DNA.- The Immunology of DNA Vaccines.- Methodology Used in DNA-Based Prophylactic and Therapeutic Immunization Against Hepatitis B Virus in Chimpanzees.- Intramuscular and Intradermal Injection of DNA Vaccines in Mice and Primates.- Veterinary DNA Vaccines.- DNA-Based Immunization of Neonatal Mice.- Intramuscular Injection of DNA Vaccines in Fish.- Development of DNA Vaccines for Salmonid Fish.- CTL Analysis for Tumor Vaccines.- The Use of Bone Marrow-Chimeric Mice in Determining the MHC Restriction of Epitope-Specific Cytotoxic T Lymphocytes.- Immunostimulatory DNA Sequences.- Immunostimulatory CpG Motifs and DNA Vaccines.- In Vitro Assay of Immunostimulatory Activities of Plasmid Vectors.- Adjuvants for Plasmid DNA Vaccines.- Cytokine and Costimulatory Factor-Encoding Plasmids as Adjuvants for DNA Vaccination.- Optimization of DNA Vaccines Through the Use of Molecular Adjuvants.- Cytokine Fusion Constructs as DNA Vaccines Against Tumors.- The Use of Conventional Immunologic Adjuvants in DNA Vaccine Preparations.- Genetic Adjuvants.- DNA Immunization in Combination with the Immunostimulant Monophosphoryl Lipid A.- Controlled Plasmid Delivery and Gene Expression.- Mucosal Immunization with DNA Vaccines.- Preparations for Particle-Mediated Gene Transfer Using the Accell® Gene Gun.- Entrapment of Plasmid DNA Vaccines into Liposomes by Dehydration/Rehydration.- DNA-Based Vaccination with Polycistronic Expression Plasmids.- A Nonviral Cytoplasmic T7 Autogene System and Its Applications in DNA Vaccination.-Immunization with Naked DNA Coexpressing Antigen and Cytokine via IRES.- Genetic Subunit Vaccines.- Antigen Engineering in DNA Immunization.- Genetic Vaccination Targeting T-Cell Receptors.- Immunity to T-Cell Receptor.- DNA Fusion Vaccines Against B-Cell Tumors.- DNA-Based Vaccination Primes Tumor-Rejecting T-Cell Responses.- Development of Female Contraceptive Vaccine Through DNA Inoculation of Human Chorionic Gonadotropin Beta Subunit (hCGß).- Genetic Immunization for Allergy Immunotherapy.- Preclinical Safety of DNA Vaccines.- DNA Vaccination.- Assuring the Quality, Safety, and Efficacy of DNA Vaccines.