Dna Vaccines

This volume details practical procedures on the latest DNA vaccine technology. Chapters guide readers through methods and protocols on DNA vaccine design, the adjuvant influence, production and purification methodologies, delivery systems, and approaches of the influence of DNA vaccines in the immunological response performance and in the cancer immunotherapy. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, DNA Vaccines: Methods and Protocols aims to ensure successful results in the further study of this vital field.

This volume details practical procedures on the latest DNA vaccine technology. Chapters guide readers through methods and protocols on DNA vaccine design, the adjuvant influence, production and purification methodologies, delivery systems, and approaches of the influence of DNA vaccines in the immunological response performance and in the cancer immunotherapy. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, DNA Vaccines: Methods and Protocols aims to ensure successful results in the further study of this vital field.

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.

In the early 1990s, almost 200 yr after Edward Jenner demonstrated the effectiveness of the smallpox vaccine, a new paradigm for vaccination emerged. The conventional method of vaccination required delivery of whole pathogens or structural subunits, but in this new approach, DNA or genetic information was administered to elicit an immunological response. Once it was observed that plasmid DNA delivered in vivo led to production of an encoded transgene (1), two ground-breaking studies demonstrated that immunological responses could be generated against antigenic transgenes via plasmid DNA delivered by DNA vaccination (as this approach is called) (2,3). The appe- ance of this new vaccination strategy coincided with advances in molecular biology, which provided new tools to study and manipulate the basic elements of an organism’s genome and also could also be applied to the design and production of DNA vaccines. DNA Vaccines is a major updated and enhancement of the first edition. It reviews state-of-the-art methods in DNA vaccine technology, with chapters describing DNA vaccine design, delivery systems, adjuvants, current appli- tions, methods of production, and quality control. Consistent with the approach of the Methods in Molecular Medicine series, these chapters contain detailed practical procedures on the latest DNA vaccine technology. The enthusiasm for DNA vaccine technology is made clear by the number of research studies published on this topic since the mid-1990s.

Genetic / DNA immunization represents a novel approach to vaccine and immune therapeutic development. The direct injec tion of nucleic acid expression cassettes into a living host results in a limited number of its cells becoming factories for production of the introduced gene products. This host-inappropriate gene expression has important immunological consequences, resulting in the specific immune activation of the host against the gene delivered antigen. The recent demonstration by a number of laboratories that the induced immune responses are functional in experimental models against both specific infectious diseases and cancers is likely to have dramatic consequences for the develop ment of a new generation of experimental vaccines and immune therapies. This technology has the potential to enable the pro duction of vaccines and immune-based therapies that are not only effective immunologically but are accessible to the entire world (rather than just to the most developed nations). Vaccine Development Vaccination against pathogenic microorganisms represents one of the most important advances in the history of medicine. Vaccines, including those against polio, measles, mumps, rubella, hepatitis A, hepatitis B, pertussis and other diseases, have dramatically improved and protected more human lives than any other avenue of modern medicine. The vaccine against smallpox, for example, has been so successful that it is now widely believed that this malicious killer, responsible for more deaths in the twentieth century than World Wars I and II combined, has been removed from the face of the earth.

This report surveys opportunities for future Army applications in biotechnology, including sensors, electronics and computers, materials, logistics, and medical therapeutics, by matching commercial trends and developments with enduring Army requirements. Several biotechnology areas are identified as important for the Army to exploit, either by direct funding of research or by indirect influence of commercial sources, to achieve significant gains in combat effectiveness before 2025.

DNA vaccination is one of the most promising techniques for immunisation against diseases caused by viruses, protozoa, bacteria, and even for tumours and illnesses with genetic origins. These vaccines can be administered by direct inoculation of plasmid by several routes. In this book, the authors present topical research in the study of the types, advantages and limitations of DNA vaccines. Topics discussed include the role of novel carrier constructs in effective and safe DNA vaccine delivery; improving the immunogenicity of DNA vaccines; the role of DNA-IL-12 vaccination in cosinphilic inflammation; genetic immunisation enabling re-direction of the host immune system resulting in the development of effective immune responses; and DNA vaccination against herpesvirus.

DNA is a rapidly developing vaccine platform for cancer and infectious and non-infectious diseases. Plasmids are used as immunogens to encode proteins to be further synthesized in vaccine recipients. DNA is mainly synthetic, ensuring enhanced expression in the cells of vaccine recipients (mostly mammalians). Their introduction into the host induces antibody and cellular responses. The latter are often more pronounced, and mimic the events occurring in infection, especially viral. There are a few distinct ways in which the vaccine antigen can be processed and presented, which determine the resulting immune response and which can be manipulated. Routinely, the antigen synthesized within the host cell is processed by proteasome, loaded onto, and presented in a complex with MHC I molecules. Processing can be re-routed to the lysosome, or immunogen can be secreted for further presentation in a complex with MHC II. Apart from expression, vaccination efficacy depends on DNA delivery. DNA immunogens are generally administered by intramuscular or intradermal injections, usually followed by electroporation, which enhances delivery 1000-fold. Other techniques are also used, such as noninvasive introduction by biojectors, skin applications with plasters and microneedles/chips, sonication, magnetofection, and even tattooing. An intense debate regarding the pros and cons of different routes of delivery is ongoing. A number of studies have compared the effect of delivery methods at the level of immunogen expression, and the magnitude and specificity of the resulting immune response. According to some, the delivery route determines immunogenic performance; according to others, it can modulate the level of response, but not its specificity or polarity. The progress of research aiming at the optimization of DNA vaccine design, delivery, and immunogenic performance has led to a marked increase in their efficacy in large species and humans. New DNA vaccines for use in the treatment of infectious diseases, cancer, allergies, and autoimmunity are forthcoming. This Special Issue covers various aspects of DNA vaccine development.