Regulatory T lymphocytes expressing CD4, high levels of CD25, and the transcription factor Foxp3 play a crucial role in the control of immune responses to self and nonself antigens. In contrast to immunosuppressive drugs currently used to treat immunopathology, these cells act in a very spec ...
The discovery of Foxp3 as a reliable marker for murine regulatory T cells has led to an explosion in the development of genetic tools for investigating the biology of regulatory T cells. More than 25 Foxp3-based mouse strains have been published with a variety of characteristics. The effects of Fox ...
Naturally occurring regulatory T (nT Reg ) cells play a critical role in the establishment of immunological self-tolerance in humans. Currently, the analysis of nT Reg cell function from bulk PBMC has led to discrepancies, largely due to the failure to discriminate T Reg cells from other antigen ...
To fully examine the functionality of a regulatory T cell (Treg) population, one needs to assess their ability to suppress in a variety of in vivo models. We describe five in vivo models that examine the suppressive capacity of Tregs upon different target cell types. The advantages and disadvantag ...
Regulatory T cells (Treg) have become increasingly relevant in the study of human disease including cancer. Treg cells have been shown to inhibit anti-tumor immune responses, and elevated Treg levels have been associated with certain types of cancer. Similarly, depletion of Tregs by vario ...
Regulatory T cells are involved in the maintenance of tolerance. Alterations in their functional capacity are implicated in the development of autoimmunity. In the case of common autoimmune disorders the defects in suppression may be partial, and may be due to a loss of Treg function, or a resista ...
The in vivo proliferation and disappearance kinetics of lymphocytes may be estimated in humans from rates of deuterium-labeled glucose (2H2-glucose) incorporation into DNA. This protocol describes its application to regulatory T cells (Treg). Because Treg divide frequently, 2H2 ...
Tregs are absolutely required for the maintenance of self tolerance in mouse and man. Major abnormalities in Treg number or function cause rare but fatal syndromes with autoimmune, allergic and inflammatory features. Whether subtle Treg abnormalities contribute to the pathogenes ...
Enzyme-linked immuno spot (ELISPOT) assay is widely used for vaccine development, cancer and AIDS research, and autoimmune disease studies. The output of ELISPOT assay is a formation of colored spots which appear at the sites of cells releasing cytokines, with each individual spot represe ...
Enzyme-linked immunospot (ELISPOT) assay allows for the determination of the frequency of �cytokine-secreting cells, but does not answer the question of how much cytokine is secreted per cell. In our study, we combined ELISPOT and ELISA assays and developed a protocol to calculate the amount ...
Adeno-associated viral (AAV) vector-mediated gene transfer represents a promising gene replacement strategy for treating various genetic diseases. One obstacle in using viral-derived vectors for in vivo gene delivery is the development of host immune responses to the vector. Rec ...
A commercial feline immunodeficiency virus (FIV) vaccine consisting of inactivated dual-subtype viruses was released in the USA in 2002 and released subsequently over the next 6 years in Canada, Australia, New Zealand, and Japan. Based on the genetic, morphologic, and biochemical simil ...
Human and mouse immune system cells are the most frequently used specimens in ELISPOT assays. In an effect to expand the application of ELISPOT assay to other species, we developed matched antibody pairs for ready-to-use kits designed for studying the frequency of equine IFNγ- and IL-4-secret ...
The T cell system plays an essential role in infections, allergic reactions, tumor and transplant rejection, as well as autoimmune diseases. It does so by the selective engagement of different antigen-specific effector cell lineages that differentially secrete cytokines and other e ...
Mesenchymal stem cells (MSCs) have many regulatory effects (e.g., T-cell suppression) on various immune cells. As aberrant T-cell activation is a primary cause of many diseases, understanding the underlying mechanisms of how MSCs exert immunosuppression is an important issue with pot ...
The major application of ELISPOT assays is to study secretion of cytokines and chemokines from immune system cells. We adapted this assay to study TNFα secretion from microglial BV2 cells, which are similar in physiology to microglia in the nervous system. Stimulation of BV2 cells with 1 μg/mL LPS r ...
Over the past decade, ELISPOT has become well-established as a mainstream technology for the study of immune responses in vivo mainly due to its unique ability to detect rare antigen-specific lymphocytes ex vivo. The primary readout for ELISPOT assays has traditionally been the measureme ...
ELISPOT results used to be evaluated visually which, however, is inevitably subjective, inaccurate, and cumbersome. Even when applying automated image analysis to this end, the results are highly variable if the counting parameters are set subjectively. Since objective, accurate, a ...
Accuracy of spot detection and quantification plays a critical role in the analysis of ELISPOT data. Differences in staining intensities of spots and their morphological variations make it difficult developing a reliable software application. We have developed an image recogniti ...
Cytokine ELISPOT assays have emerged as a powerful tool for the detection of rare antigen-specific T cells in freshly isolated cell material, such as blood. While ELISPOT assays allow one to directly visualize and count extremely low frequencies of cytokine-secreting T cells among millio ...