遗传学实验技术

The biochemical and genetic properties of DNA have been thoroughly investigated, yet only recently has it been appreciated that DNA carries more information than simply a blueprint for the regulation and construction of proteins. Indeed, the immune systems of vertebrates appear to have ...

Plasmid DNA immunization has emerged as a promising vaccine strategy against infectious agents, as well as a potential intervention for the treatment of cancer, autoimmunity, and allergy (1). Until recently, however, the cellular events by which injected plasmid DNA elicits potent ant ...

The DNA-based immunization technology has only been applied to fish very recently. Though a preliminary study showing reporter gene expression in fish muscles was published in 1991 (1), the first demonstration of an immune response to plasmid-encoded antigen was not reported until five y ...

Immunization of neonates carries three inherent questions: (i) Is the neonatal immune system mature enough to elicit an immune response, and if not, how early can immunization be carried out? (ii) If an immune response is not elicited, will tolerance be induced? (iii) Can offspring of immune mothe ...

Vaccination has relied, in general, on two technologies for the production of antigenic material suitable for the generation of a protective immune response; live infectious agents and inactivated or subunit vaccine preparations. Live infectious agents generally are most effica ...

There are several different ways to administer plasmid DNA vaccines (1). Those most commonly used include intramuscular (i.m.) injection or intradermal (i.d.) injection of pure plasmid DNA (“naked” DNA), or biolistic introduction of DNA-coated gold particles into the epidermis with a “ge ...

Chimpanzees, because of their near-human nature, have special needs that must be met by those who carry out medical research with them. Perhaps the most important of these is the need for companionship. Chimpanzees kept alone become obviously depressed, and manifest stereotypic behavior s ...

The surprising observation that direct inoculation of an expression plasmid encoding a foreign protein into the skin of mice resulted in the induction of antibody responses, demonstrated that injection of “naked” DNA could result in antigen expression in an immunogenic form (1). This obs ...

The preparation of large amounts of high-purity intact plasmid DNA is a significant expense in DNA vaccine research. In our laboratory, mice are typically each immunized by injection of 100 μg on four occasions, so that an experiment with 50 mice requires 20 mg DNA as a minimum. The Qiagen tip-10,000 (Gi ...

Scientists in academia whose research is aimed at the development of a novel vaccine or approach to vaccination may not always be fully aware of the regulatory process by which a candidate vaccine becomes a licensed product. This chapter will provide an overview of the regulatory process and will ...

Infectious diseases result in significant morbidity and mortality worldwide. Preventing these infections is both a public health priority and the primary goal of vaccine research. The discovery that cell-mediated and/or humoral immune responses against viruses, parasites, ba ...

The potential applications of using plasmid DNA for immunization and other gene therapy approaches have been discussed in an increasing number of publications in the past few years. Injection of mouse muscle with naked DNA (plasmid DNA in saline) resulted in significant episomal express ...

DNA or genetic vaccines are currently being evaluated for safety and efficacy in human clinical trials in the areas of infectious disease and cancer. Since DNA vaccines induce antibodies and cytotoxic T lymphocytes (CTLs), they are currently being evaluated in humans for both prevention a ...

Allergic disorders are characterized by the prevalence of immunoglobulin (Ig) isotype E antibodies, and are considered to result from enhanced T helper type-2 (Th2) responses to allergens. A Th2 response is characterized by enhanced humoral responses and the production of IL-4 and IL-5 by C ...

Human chorionic gonadotropin (hCG) has been considered as a primary target molecule for a contraceptive vaccine by the World Health Organization because of its physiological and temporal specificity. hCG is an essential factor for the successful implantation and establishment of e ...

DNA-based vaccination efficiently primes MHC-restricted T-cell responses. This technique specifically stimulates MHC-II-restricted CD4+ T-cell responses and MHC-I-restricted CD8+ T-cell responses against “strong” (immunodominant) or “weak” (subdominant or crypt ...

The ability of naked DNA to induce immune responses against encoded antigen has been clearly demonstrated for infectious diseases (1). In many cases, the induced immunity is able to protect against infection, and can approach the efficacy of exogenous antigen (2).

A common theme found in studies with mouse models of autoimmune diseases is that pathogenic T cells are primarily responsible for the pathology. Such models include diabetes in non-obese diabetic (NOD) mice, experimental autoimmune encephalomyelitis (EAE)—a model of multiple scler ...

T-cell antigen receptor (TCR) genes (which consist of variable (V), diversity (D), joining (J) and constant (C) segments) undergo rearrangement during T-cell development and result in the expression of a disulfide linked heterodimer (α and � chains) on the surface of mature T-cells (1,2). The TCR c ...

J. A. Wolff and colleagues demonstrated in 1990 (1) that naked, non-infectious DNA plasmids can be transferred into muscle cells of living mice to express the encoded protein in vivo. Further studies have shown that proteins expressed in vivo via such a “DNA inoculation” process can serve as immuno ...