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Bacteria-Mediated Acute Lung Inflammation

Mouse models of acute lung inflammation are critical for understanding the role of the innate immune response to pathogen associated molecular patterns, bacteria, and sepsis in humans. Bacterial infections in the lung elicit a range of immune reactions, depending on the pathogen, the lev ...

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Endotoxin-Induced Uveitis in Rodents

Uveitis is a common cause of vision loss, accounting for 10–15 % of all cases of blindness worldwide and affects individuals of all ages, genders, and races. Uveitis represents a broad range of intraocular inflammatory conditions due to complications of autoimmune diseases, bacterial infe ...

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Mouse Model of Invasive Fungal Infection

The mouse intravenous (IV) challenge model of Candida albicans invasive fungal infection has been widely used to study the importance of the innate immune system in these infections. This chapter describes this well-characterized model, where fungal cells are administered directly ...

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Systemic Infection of Mice with Listeria monocytogenes to Characterize Host Immune Responses

Listeria monocytogenes is a Gram-positive facultative intracellular bacterium that is widely used to characterize bacterial pathogenesis and host immunity. Here, we describe a set of basic methods and techniques to infect mice with L. monocytogenes, measure bacterial load in tiss ...

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Sepsis Induced by Cecal Ligation and Puncture

Despite advances in intensive care unit interventions, including the use of specific antibiotics and anti-inflammation treatment, sepsis with concomitant multiple organ failure is the most common cause of death in many acute care units. In order to understand the mechanisms of clinic ...

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Mouse Model of Staphylococcus aureus Skin Infection

Bacterial skin and soft tissue infections are abundant worldwide and many are caused by Staphylococcus aureus. Indeed, S. aureus is the leading cause of skin and soft tissue infections in the USA. Here, we describe a mouse model of skin and soft tissue infection induced by subcutaneous inoculati ...

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Delayed-Type Hypersensitivity Models in Mice

Delayed-type hypersensitivity (DTH) is a useful approach for evaluating cell-mediated immune responses associated with Th1 reactivity. The DTH reaction is divided into the afferent and efferent phases. During the afferent phase of this model, mice are typically immunized by subcut ...

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Safe and Effective Mouse Footpad Inoculation

Footpad injection is an important route of inoculation in mouse models of disease and immunology. Although commonly used to deliver antigens as a means of eliciting an efficacious immunological response, herein, we describe a protocol for inoculating mice via footpad injection using a h ...

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Bioassay for the Measurement of Type-I Interferon Activity

Type I interferons are critical cytokines produced by the host innate immune response to viral infection. They act collectively to initiate expression of a multitude of antiviral genes that serve to inhibit viral replication and spread. Despite the great importance of interferons to the h ...

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Measuring T Cell Function in Innate Immune Models

Innate immune responses often result in the activation and modulation of T lymphocyte function. Analysis of T lymphocytes in mouse models of innate immunity can allow understanding of the links between the innate and adaptive immune systems. Other T lymphocyte populations display inna ...

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Homeostatic Proliferation of Mature T Cells

Under normal circumstances, the secondary lymphoid tissues contain a predictable number of T cells with a diverse T cell receptor (TCR) repertoire. Such a T cell pool must be of sufficient size to confer maximum protection of the host from infectious pathogens and cancer, but small enough not to ove ...

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Assessment of CD4+ and CD8+ T Cell Responses Using MHC Class I and II Tetramers

The low frequency of T cells specific for given antigens makes the study of antigen-specific T cell responses difficult. The development of MHC class I and II tetramer staining techniques allows precise quantification and tracking of antigen-specific CD8+ and CD4+ T cell responses. Here, we d ...

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Detection of Necrosis by Release of Lactate Dehydrogenase Activity

Apoptosis and necrosis are two major forms of cell death observed in normal and disease pathologies. Although there are many assays for detection of apoptosis, relatively few assays are available for measuring necrosis. A key signature for necrotic cells is the permeabilization of the pla ...

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Analyses of Programmed Cell Death in Dendritic Cells

Dendritic cells (DCs) are professional antigen-presenting cells that can regulate both innate and adaptive immune responses. Programmed cell death of DCs plays an important role in maintaining the homeostasis of DCs and in the regulation of immune responses. Methods to measure the rate of ...

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Visualization of Fas-Mediated Death-Inducing Signaling Complex Formation by Immunoprecipitation

The prototypical death receptor Fas (also known as CD95 or Apo-1) plays an essential role in the maintenance of lymphocyte homeostasis. Propagation of cell death through Fas relies on the formation of a multiprotein complex at the receptor level known as the death-inducing signaling complex ...

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Determination of Apoptosis Sensitivity in Specific T Cell Subsets from Human Peripheral Blood by Utilizing a Multiparameter Fluorescence-Activated Cel

Among the different techniques available for determining physiological cell death or apoptosis in immune cells, fluorescence-activated cell sorting-based approaches prove to be one of the most efficient and quantitative assays in capturing cells that are actively undergoing ...

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Evaluation of IL-2-Withdrawal-Induced Apoptosis in Human T Lymphocytes

Proper lymphocyte apoptosis is critical for the maintenance of immune system homeostasis, and evaluation of cell death is useful in a number of clinical and research settings. We describe here how to evaluate the integrity of the intrinsic pathway of lymphocyte apoptosis triggered by star ...

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Fluorescence-Activated Cell Sorting-Based Quantitation of T Cell Receptor Restimulation-Induced Cell Death in Activated, Primary Human T Cells

After initial stimulation with antigen and exposure to the growth cytokine interleukin-2, activated T lymphocytes become sensitized to apoptosis upon antigen restimulation through the T cell receptor. This self-regulatory, restimulation-induced cell death (RICD) program ...

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Quantitating Lymphocyte Programmed Cell Death In Vitro Using Simple Kill Assays

Programmed cell death is essential to maintaining lymphocyte homeostasis during the contraction phase of the immune response. Activated lymphocytes become susceptible to a variety of programmed cell death (PCD) stimuli over the course of a typical immune response. This chapter out ...

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In Vitro Suppression Assay for Functional Assessment of Human Regulatory T Cells

FOXP3+ regulatory T cells (Tregs) play an important role in controlling immune activation, maintenance of �homeostasis and the prevention of autoimmunity. Much effort has been focused in assessing their potential defects in certain human diseases and in developing potential Treg im ...

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