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Drosophila Immunity: Methods for Monitoring the Activity of Toll and Imd Signaling Pathways

Invertebrates lack an adaptive immune system and rely on innate immunity to resist pathogens. The response of Drosophila melanogaster to bacterial and fungal infections involves two signaling pathways, Toll and Imd, both of which activate members of the nuclear factor (NF)-κB family of t ...

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Investigating the Involvement of Host Factors Involved in Intracellular Pathogen Infection by RNAi in Drosophila Cells

Intracellular pathogens represent a serious threat to human health. Although the biology and the virulence factors involved in intracellular bacterial infection are relatively well documented, little is known about the host factors involved in the infection process. This situa ...

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Genetic Analysis of Caenorhabditis elegans Innate Immunity

Innate immunity is an ancient and conserved defense mechanism. The worm Caenorhabditis elegans provides a useful tool for studying the function of the innate immune system at the molecular and cellular levels within the context of a whole organism. The powerful genetics of the worm, combined ...

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Models of Caenorhabditis elegans Infection by Bacterial and Fungal Pathogens

The nematode Caenorhabditis elegans is a simple model host for studying the relationship between the animal innate immune system and a variety of bacterial and fungal pathogens. Extensive genetic and molecular tools are available in C. elegans, facilitating an in-depth analysis of host d ...

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Measuring Cell-Wall-Based Defenses and Their Effect on Bacterial Growth in Arabidopsis

Plants are resistant to most potentially pathogenic microbes. Frequently, resistance results from defenses activated upon recognition of “non-self.” Invasion of a variety of pathogens, including Gram-negative bacteria, into plants is betrayed by the presence of pathogen-ass ...

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Innate Immunity Genes as Candidate Genes: Searching for Relevant Natural Polymorphisms in Databases and Assessing Family-Based Association of Polymorp

The identification of genes underlying complex traits is a challenging task, and there are a limited number of confirmed genes that influence human complex diseases. In particular, few genes involved in complex diseases related to immune response, such as infectious diseases and inflam ...

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KIR Genotyping and Analysis: Genotyping and Disease Association Analysis

The genes encoding the killer immunoglobulin-like receptors (KIR) are situated within a segment of DNA that has undergone expansion and contraction over time due in large part to unequal crossing over. Consequently, individuals exhibit considerable haplotypic variation in terms of ...

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Bacterial Artificial Chromosome Transgenesis Through Pronuclear Injection of Fertilized Mouse Oocytes

In the mouse, conventional transgenes often produced unpredictable results mainly because they were too small to recapitulate a natural gene context. Bacterial artificial chromosomes (BACs) are large enough to encompass the natural context of most mammalian genes and consequen ...

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Bioluminescence Imaging to Evaluate Infections and Host Response In Vivo

The continued prospect of emerging pathogens and recent events including the acceptance of widespread drug resistance and threats of bioterrorism have introduced the necessity be creative in our development of therapies for bacterial infections. Many pathogens have both acute a ...

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Intravital Two-Photon Imaging of Natural Killer Cells and Dendritic Cells in Lymph Nodes

Two-photon microscopy makes it possible to image in real-time fluorescently labeled cells located in deep tissue environments. We describe a procedure to visualize the behavior of natural killer (NK) cells and dendritic cells (DC) in the lymph nodes of live, anesthetized mice. Intravital ...

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Dissection of the Antiviral NK Cell Response by MCMV Mutants

Our understanding of virus control by natural killer (NK) cells relies mainly on in vitro observations. The significance of these findings for virus control in vivo is not yet fully understood. Complexity is added by the fact that many viruses, particularly herpesviruses, are equipped with s ...

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Analyzing AntibodyFc-Receptor Interactions

Cellular receptors for immunoglobulins (Fc-receptors; FcR) are central mediators of antibody-triggered effector functions. Immune complex (IC) binding to FcRs results in a variety of reactions such as the release of inflammatory mediators, antibody dependent cellular cytot ...

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The Use of Adjuvants in Experimental Vaccines: II. Water-in-Oil Emulsions: Freunds Complete and lncomplete Adjuvants

As mentioned in Chapter 9, Freund’s complete adjuvant (FCA), produced by the Statens Seruminstitut (Copenhagen, Denmark), was chosen as the standard adjuvant preparation because of its extensive use in experimental vaccines in animals, its strong adjuvant effect, and the considerab ...

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The Use of Adjuvants in Experimental Vaccines: III. Liposomes

The use of artificial lipid bilayers in the form of vesicles has been described as an efficient means of presenting antigens (1, 2 and Chapter 9). A large variety of natural phospholipids or other polar amphiphiles can be used in an aqueous solution of an antigen to form either unilamellar or multilame ...

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The Use of Adjuvants in Experimental Vaccines: IV. ISCOMS

The use of saponin in experimental vaccines has been known for more than 60 yr (1, Chapter 9) and generally it is more active as an adjuvant with strongly immunogenic antigens. A number of saponins are derived from the bark of the South American soaptree (Quillaja saponaria) as acylated triterpenoids ...

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Microencapsulation of Vaccines

Despite obvious successes in controlling most serious childhood infections, there is constant pressure to develop cheaper, safer, and more effective infant vaccination programs. However, any improvements to pediatric vaccines in the foreseeable future are likely to arise thro ...

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Stimulation of Mucosal Immunity

The mucosal immune system is composed of distinct regional immune tissue (e.g., “GALT,” gut-associated lymphoid tissue; “BALT,” bronchus-associated lymphoid tissue; reproductive tract and breast tissue, and so forth) interconnected by trafficking of primed lymphocytes as a comm ...

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Lyophilization of Vaccines

However effective a therapeutic agent may appear during clinical trials, its full potential as a marketable product can only be assured if the material can be stored and distributed in a stabilized form. This is particularly the case with labile biologlcals, such as attenuated vaccines, espe ...

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Induction and Detection of T-Cell Responses

The fundamental basis of immunity to infectious diseases involves a complex interplay between humoral and cell-mediated immune responses against antigens on the foreign pathogen. Humoral immunity, either in the form of local IgA antibodies at the mucosal site of infection or neutral ...

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Immunological Characterization

As part of the assessment of product consistency and putative potency, it is often necessary to check not only that a vaccine contains the desired antigens, but also that they are in the correct immunological form (see Note 1). In general, this immunological characterization of a vaccine is interp ...

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