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Experimental Strategies for Combined Suicide and Immune Cancer Gene Therapy: An Overview

Suicide gene therapy systems are characterized by the transfer of therapeutic transgenes, which encode for enzymes of various origins. These enzymes are able to convert nontoxic prodrugs into highly cytotoxic metabolites. Thus, all cells that are transduced by suicide genes will be des ...

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Gene Therapy with Plasmids Encoding Cytokine- or Cytokine Receptor-IgG Chimeric Proteins

Cytokine therapy can influence the outcome of autoimmune diseases by altering either T-helper 1 (Th1) vs T-helper 2 (Th2) balance or antigen-presenting cell (APC) function, or by shifting the balance between inflammatory and regulatory cytokines (1). However, cytokine and soluble cyto ...

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Interferon- Gene Therapy for the Treatment of Arthritis

Interferons were originally described as antiviral agents by Isaacs and Lindenmann (1). From its subtypes, fibroblast interferon (IFN-β) (2) was also found to have important cytostatic (3,4) as well as immunomodulatory functions (5) that have been advantageous for its clinical use both in ...

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Co-Culture of Mast Cells With Fibroblasts: A Tool to Study Their Crosstalk

Mast cell development, function, and survival are likely to be regulated by a complex interplay of cellular signaling. Usually, these signals derive from the cellular milieu associated with the specific mast cell environment in health or disease conditions. A major methodological iss ...

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Detection of Class Switching and IgE Synthesis in Human B Cells

We observed that mast cells, as other cells expressing the CD40 ligand CD154, can trigger IgE synthesis in B cells in the presence of interleukin (IL)-4. Numerous complementary techniques can be used to follow the succession of molecular events leading to IgE synthesis. This chapter will illust ...

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Gene Silencing Using Small Interference RNA in Mast Cells

Small interfering RNA (siRNA) is a potent and specific method of inducing gene silencing through induction of RNA interference. siRNAs can be allowed for in vitro and in vivo applications. siRNAs have been successfully studied in vitro, but little is known about its efficacy in vivo. We have succe ...

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Mast Cell Activation by Stress

Mental or emotional stress has been shown to cause mast cell degranulation in several different tissues. Several lines of experimental evidence indicate that stress, working through the sympathetic nervous system, or the hypothalamus-pituitary-adrenal axis, stimulates peri ...

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Mast Cell Activation by Lipoproteins

Mast cells are activated by a number of agents that act independently from immunoglobulin E (IgE)-mediated hypersensitivity. One of these agents is oxidized low-density lipoprotein (oxLDL). OxLDL has been implicated in the pathogenesis of atherosclerosis and has been shown to induce m ...

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In Vivo Models for Studying Mast Cell-Dependent Responses to Bacterial Infection

Mast cells are a critical component of host defense against bacterial infections. Activation of these cells during infection induces both innate and adaptive aspects of protective immunity needed for the elimination of the bacteria and survival of the host. These functional roles for the ...

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Bacterial Activation of Mast Cells

Mast cells often are found in a perivascular location but especially in mucosae, where they may response to various stimuli. They typically associate with immediate hypersensitive responses and are likely to play a critical role in host defense. In this chapter, a common airway pathogen, Mor ...

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Activation of Mast Cells by Streptolysin O and Lipopolysaccharide

This chapter provides protocols to measure the reversible permeabilization of mast cells by streptolysin O (SLO) and to follow SLO-induced activation of mast cells by monitoring degranulation, activation of mitogen-activated protein kinases, and production of tumor necrosis f ...

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Mast Cell Apoptosis

Apoptosis is a physiological form of cell death. Cells undergoing apoptosis execute a genetically controlled program that leads to organized breakdown of cellular structures and ends in phagocytosis of their remains. In mast cells, several mechanisms regulating apoptosis have be ...

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Purification of the Eucaryotic Heat-Shock Proteins Hsp70 and gp96

Heat-shock proteins (HSPs), highly conserved across species, are generally considered as intracellular proteins that have protective functions in situations of cellular stress. A wide variety of stressful stimuli like heat shock, ultraviolet radiation, and viral or bacterial i ...

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Genetic Engineering of a Recombinant Fusion Protein Possessing an Antitumor Antibody Fragment and a TNF- Moiety

Tumor necrosis factor-α (TNF-α) is a cytokine (CK) that possesses a wide variety of biological activities, including potent antitumor activities (1) and immunomodulatory properties mediated through its binding to two TNF receptors (p55 and p75) (2). Signaling through the p55 receptor is ...

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Genetic Engineering of Dendritic Cells by Adenovirus-Mediated TNF- Gene Transfer

Dendritic cells (DCs) are one of the most potent antigen-presenting cells (APCs). They migrate as precursors from the bone marrow into various organs, where they usually reside in an inactive state (1). However, during this regional residency, these cells can efficiently endocytose and pro ...

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Retroviral Transfer of T-Cell Receptor Genes into Human Peripheral Blood Lymphocytes

We are developing strategies to target tumor-associated antigens (TAA) and viral antigens by genetically modifying patient peripheral blood lymphocytes to produce antitumor or antiviral reactive cytotoxic T-lymphocyte (CTL). To this end, we have succeeded in redirecting the sp ...

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Gene Transfer to Articular Chondrocytes with Recombinant Adenovirus

Articular cartilage forms the load-bearing surfaces of diarthrodial joints. Its dense extracellular matrix, composed primarily of water, collagen, and proteoglycans is maintained by a resident population of chondrocytes. These all combine to form a highly organized structure t ...

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Gene Therapy of X-Linked Severe Combined Immunodeficiency

This review describes the main steps which led to carrying out the gene ther-apy clinical trial for Xlinked severe combined immune deficiency (SCID-X1) patients, from the preparation of the retroviral vector up to the design of GMP conditions to transduce ex vivo the CD34+cells of patients. This ...

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Gene Transfer for Generation of Tumor and Leukemia Vaccines

Tumor vaccine strategies aim to compensate for the reduced immunogenicity of tumor cells (1-4). Neoplastic cells fail to induce an effective antineoplastic immune response for a number of reasons, such as defective antigen-processing mechanisms, decreased levels of MHC expressio ...

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In Vitro Regulation of Colony Stimulating Factor-Mediated Hematopoiesis in Healthy Individuals and Patients with Different Types of Myeloproliferative

Human hematopoiesis is initiated by hematopoietic stem cells, the most undifferentiated cell type in the bone marrow. Although hematopoietic stem cells are not yet fully identified, their function is well characterized. They are able to replicate by cell division producing daughter c ...

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