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 ...

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 ...

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 ...

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 ...

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 ...

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 ...

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 ...

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 ...

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 ...

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 ...

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 ...

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 ...

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 ...

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 ...

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 ...

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 ...

Flow cytometry is a valuable tool for the detection and characterization of proteins expressed by individual cells. Flow cytometry can be used to measure cell expression of 2 intracellular proteins that are involved in the regulation of immune homeostasis, SLAM-associated protein (S ...

Massively parallel sequencing technologies provide new opportunities to discover causal variants and �narrow down candidate genes responsible for human Mendelian disorders. Such information can in turn provide new insights into understanding the basic science behind, as we ...

Genome-wide gene expression analysis has become a very powerful routine tool for the study of distinct differentiation states. However, the examination of total populations of cells that contain high levels of heterogeneity, such as the total CD8+ T cell population during an immune respo ...

This chapter provides protocols necessary for quantifying human, mouse, and nonhuman primate signal joint T cell receptor excision circles (sjTRECs) produced during T cell receptor alpha (TCRA) gene rearrangement. These non-replicated episomal circles of DNA are generated by the r ...