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丁香实验推荐阅读
Methods of Immunization to Enhance the Immune Response to Specific Antigens In Vivo in Preparation for Fusions Yielding Monoclonal Antibodies

The first step in preparing useful monoclonal antibodies (MAbs) is to immunize an animal with an appropriate “vaccine.” Animal and vaccine are both emphasized in the preceding sentence because this chapter describes how to generate satisfactory MAbs by maximizing interactions betw ...

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Scintillation Proximity Binding Assay

Receptor-ligand interactions have been extensively used to establish the specificity and binding profiles for chemokine receptors and their ligands. Furthermore, receptor-ligand binding assays are used extensively for drug screening. The Scintillation Proximity Assay ...

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Rabbit Models of Pneumonia, Peritoneal Sepsis, and Lung Injury

The Acute Respiratory Distress Syndrome (ARDS) is a form of lung injury characterized by the rapid onset of severe hypoxemia and diffuse pulmonary infiltrates, with normal cardiac filling pressures (1). The pathology of ARDS includes striking injury to the alveolar epithelium, and the ac ...

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Endothelial Cell Activation by Mast Cell Mediators

Mast cells are important cells of the immune system, and their secretory products regulate many vascular functions. Although considerable interest is focused on the role of mast cells and infectious agents in atherosclerosis, whether or not mast cell mediators act in concert with bacter ...

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Gene Gun-Based In Vivo Gene Transfer: Application to DNA Vaccination

The introduction of genetic material into tissue of interest remains a ratelimiting step for molecular investigations in many fields. Many different methods have been developed for the delivery of a gene in vivo, such as virus-, cationic liposome-, and particle-mediated gene transfer or d ...

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Production of Chimeric Heavy-Chain Antibodies

Antibody has become a major category of therapeutics. However, IgG, the primary molecular format of existing antibody drugs, has some major shortcomings such as undesirable pharmacokinetics, high dose requirement, and high production cost, partially due to its large molecular size. M ...

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The Isolation and Identification of Murine Dendritic Cell Populations from Lymphoid Tissues and Their Production in Culture

Dendritic cells (DC) are widely regarded as the most potent cellular inducers of the adaptive immune response; so, immunotherapy through DC manipulation is a promising option in the future fight against many human ailments. We have developed a method of isolating DC from the mouse that involves ...

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Isolation and Analysis of Human Natural Killer Cell Subsets

Natural killer (NK) cells were originally defined as mediators of spontaneous cytotoxicity against virus-infected and tumor cells. In human peripheral blood, the majority of NK cells can mediate cell lysis mainly through perforin and granzymes. It has been recently shown, however, that ...

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Analysis of Individual Natural Killer Cell Responses

Typical assays for natural killer (NK) cell function assess the responses of entire NK cell populations. It is now possible to determine the responses of individual NK cells. Herein, two representative assays are described along with examples of how they have helped clarify current underst ...

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Innate Immune Function of Eosinophils: From Antiparasite to Antitumor Cells

Eosinophils are multifunctional leukocytes classically described as being involved in helminth parasitic infections and allergic diseases. Previously restricted to an exclusive role in the release of cytotoxic mediators, they are now also considered to be immunoregulatory ...

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Ex Vivo and In Vitro Primary Mast Cells

Mast cells are cells of the innate immune system whose biological responses are markedly modulated by effector molecules of adaptive immunity, i.e., antibodies. They thus contribute to anti-infectious defense but also to antibody-dependent inflammatory responses. They are espec ...

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Clinical Analysis of Dendritic Cell Subsets: The Dendritogram

Dendritic cells (DCs) are crucial in adaptive immunity because they are the only antigen-presenting cells that can present antigens to naive T lymphocytes. Plasmacytoid DCs (pDC) are also the main producers of type I Interferons in response to infection. We have shown that circulating myel ...

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Murine Macrophages: A Technical Approach

In this chapter, we describe current protocols used for the characterization of macrophages (MΦ) in mouse tissues and in cell suspensions from spleen and lymph nodes. Also, we include a brief description of a complementary approach: culture of primary MΦ. Although culture MΦ are extremely use ...

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Lentiviral Transduction of Immune Cells

Gene transfer into mammalian cells has been of crucial importance for studies determining the role of specific genes in the differentiation and cell fate of various hematopoietic lineages. Until recently, the majority of these studies were performed in transformed cell lines due to diff ...

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Clinical Analysis of Human Natural Killer Cells

Natural killer (NK) cells are major actors of innate immune responses against viruses, bacteria, parasites, and other mediators of pathology such as malignant transformation. These cells are also directly implicated in the link between innate and adaptive immunity, shaping T-cell re ...

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ENU Mutagenesis in Mice

Forward genetics has led to many “breakthrough” discoveries, and with the mouse genome almost fully sequenced, the creation of phenotypes through random germline mutagenesis has become an efficient means by which to find the function of yet undescribed genes. In this chapter, we will provi ...

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Axenic Mice Model

The interaction of an organism with surrounding microbial communities has a profound impact on its anatomy, physiology, and behavior. The innate immune system plays a major role in the crosstalk between bacteria and the hosts they colonize. Axenic mice provide a powerful in vivo controlled m ...

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Reverse Genetics Analysis of Antiparasitic Responses in the Malaria Vector, Anopheles gambiae

Anopheles mosquitoes are the major vectors of human malaria parasites. Mosquito–parasite interactions are critical for disease transmission and therefore represent a potential target for malaria control strategies. Mosquitoes mount potent antiparasitic responses, and ...

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In Vivo Analysis of Zebrafish Innate Immunity

Among vertebrate model species, the zebrafish embryo combines at an unprecedented level optical accessibility with easy genetic manipulation. As such, it is gaining recognition as a powerful model to study innate immunity. In this chapter, we provide a protocol for the generation of zebr ...

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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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