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Humanized Mice for Studying Human Leukocyte Integrins In Vivo

Humanized mice have recently emerged as powerful translational animal models for studying human hematopoiesis, immune interactions, and diseases of the human immune system. Several important advances in the humanized mouse technology have been reported over the last few years, th ...

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Integrin-Targeted Nanoparticles for siRNA Delivery

Integrins are heterodimeric membrane glycoproteins composed of noncovalently associated α and β subunits. Integrins support cell attachment and migration on the surrounding extracellular matrix as well as mediate cell–cell interaction in physiological and pathological ...

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PET-Radioimmunodetection of Integrins: Imaging Acute Colitis Using a 64Cu-Labeled Anti-7 Integrin Antibody

Integrins are involved in a wide range of cell interactions. Imaging their distribution using high-resolution noninvasive techniques that are directly translatable to the clinic can provide new insights into disease processes and presents the opportunity to directly monitor new ...

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Evaluating Posttranscriptional Regulation of Cytokine Genes

A wide variety of cytokines are necessary for cell–cell communication in multicellular organisms, and cytokine dysregulation has detrimental effects, leading to disease states. Thus, it is a necessity that the expression of cytokines is tightly controlled. Regulation of cytokine g ...

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Quantitative Analysis of miRNA Expression in Epithelial Cells and Tissues

Reliable detection of the microRNA (miRNA) precursor and mature form expression levels is a fundamental starting block for more focused studies of the biogenesis and functional roles of these important post-transcriptional modulators of gene expression. Building on our experti ...

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Interleukin-27 Induces Interferon-Inducible Genes: Analysis of Gene Expression Profiles Using Affymetrix Microarray and DAVID

We have previously demonstrated that IL-27 is a novel anti-HIV cytokine, which inhibits HIV replication in CD4 T cells and macrophages as interferon (IFN)-α does. To further understand the mechanism of the antiviral effect, we performed Affymetrix DNA microarray and gene functional anno ...

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The Use of Real-Time Quantitative PCR for the Analysis of Cytokine mRNA Levels

Over the last decade, real-time-quantitative PCR (RT-qPCR) analysis has become the method of choice not only for quantitative and accurate measurement of mRNA expression levels, but also for sensitive detection of rare or mutated DNA species in diagnostic research. RT-qPCR is based on the st ...

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A Tale of Two Interferon Bioassays: How Frustration with Discrepant Results from Slightly Dissimilar Methods Can Engender Discovery

This introductory article describes an episode that took place in the mid-1980s when the first wave of cytokine discoveries took place. During studies aimed at complete purification of human interferon-γ from crude mitogen-stimulated lymphokine preparations, the use of two differ ...

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A Sensitive and Versatile Cytokine Bioassay Based on Type I Interferon Signaling in 2fTGH Cells

We have designed a sensitive and versatile bioassay for quantification of series of cytokines. The assay makes use of chimeric receptors composed of the extracellular, ligand-binding part of the cognate cytokine receptor and the transmembrane and cytosolic part of the type I interferon ...

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In Vitro Stimulation and Detection of IFN Production in Human Plasmacytoid Dendritic Cells

Type-1 interferons (IFNs), including IFNα/β, are a family of cytokines produced rapidly upon pathogen encounter and crucial for bridging innate and adaptive immunity. IFNα has been widely appreciated as a multifunctional cytokine involved particularly in early immune responses a ...

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Ligand Affinity Chromatography, an Indispensable Method for the Purification of Soluble Cytokine Receptors and Binding Proteins

Ligand affinity chromatography separation is based on unique interaction between the target analyte and a ligand, which is coupled covalently to a resin. It is a simple, rapid, selective, and efficient purification procedure of proteins providing tens of thousands fold purification in ...

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Use of RNA Interference to Investigate Cytokine Signal Transduction in Pancreatic Beta Cells

Type 1 diabetes (T1D) is a chronic autoimmune disease characterized by immune infiltration of the pancreatic islets resulting in an inflammatory reaction named insulitis and subsequent beta cell apoptosis. During the course of insulitis beta cell death is probably caused by direct con ...

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shRNA-Induced Interferon-Stimulated Gene Analysis

RNA interference (RNAi) is a cellular mechanism to inhibit the expression of gene products in a highly specific manner. In recent years, RNAi has become the cornerstone of gene function studies, shortening the otherwise long process of target identification and validation. In addition, sm ...

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Lentiviral Vector-Mediated Transgenesis in Human Embryonic Stem Cells

Human Embryonic stem cells (hESCs) offer an invaluable tool for revealing human biology and a potential source of functional cells/tissues for regenerative medicine. The utility of hESCs will likely be significantly enhanced and broadened by our ability to build versatile genetica ...

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Lentiviral Vector Transduction of Dendritic Cells for Novel Vaccine Strategies

Dendritic cells (DCs) are key antigen-presenting cells that induce primary and memory immune response. Patients with chronic infections or cancer often display DC dysfunctions. Modification of DCs or DC progenitors in vitro may overcome the problems with defective DCs in vivo. Lentiv ...

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Manipulating the Cell Differentiation Through Lentiviral Vectors

The manipulation of cell differentiation is important to create new sources for the treatment of degenerative diseases or solve cell depletion after aggressive therapy against cancer. In this chapter, the use of a tissue-specific promoter lentiviral vector to obtain a myocardial pure ...

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Lentivirus-Based Virus-Like Particles as a New Protein Delivery Tool

Virus Like Particles (VLPs) are self-assembling, nonreplicating, nonpathogenic, genomeless particles similar in size and conformation to intact infectious virions. The possibility of engineering VLPs to incorporate heterologous polypeptides/proteins renders VLPs ...

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Integrase Defective, Nonintegrating Lentiviral Vectors

Lentiviral vectors are a powerful tool for gene transfer into target cells in vitro and in vivo. However, there are concerns about safety with regard to their use in gene transfer protocols because of insertional mutagenesis following viral infection. Once in the target cells, and in addition to t ...

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Doxycycline Regulated Lentiviral Vectors

Lentiviral vectors are a powerful tool to achieve regulated expression of transgenes in vivo and in vitro. The doxycycline-inducible system is well characterized and can be used to regulate expression mediated by lentiviral vectors. Because many different doxycycline-inducible ...

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Strategies to Insulate Lentiviral Vector-Expressed Transgenes

Lentiviruses are capable of infecting many cells irrespective of their cycling status, stably inserting DNA copies of the viral RNA genomes into host chromosomes. This property has led to the development of lentiviral vectors for high-efficiency gene transfer to a wide variety of cell typ ...

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