Trans-vivo Dth Assay
Trans-vivo Dth Assay - Related Videos
11:49
Trans-vivo Delayed Type Hypersensitivity Assay for Antigen Specific Regulation
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Cited by 19 •
2013
We describe a valuable diagnostic assay that could potentially be used to decide the withdrawal of immunosuppression after transplant without elevated risk of graft rejection. The assay uses the principles of Delayed Type Hypersensitivity and provides accurate assessment of both donor specific effector and regulatory immune responses mounted by recipients.
13:26
Induction and Monitoring of Active Delayed Type Hypersensitivity (DTH) in Rats
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Cited by 18 •
2007
Delayed type hypersensitivity (DTH) is an inflammatory reaction mediated by CCR7- effector memory T lymphocytes. Here we demonstrate how to induce active DTH in Lewis rats and monitor the inflammatory response.
09:59
Imaging Effector Memory T cells in the Ear After Induction of Adoptive DTH
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Cited by 2 •
2008
Here we demonstrate a method for inducing and recording the progress of a delayed type-hypersensitivity (DTH) reaction in the rat ear. This is followed by a demonstration of the preparation of rat ear tissue for two-photon imaging of the effector / memory T cell response.
04:16
Cell Aggregation Assay to Detect Trans Interactions between Cell Adhesion Molecules
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2025
In this video, we demonstrate the technique to measure transcellular interactions between synaptic cell adhesion molecules through the HEK cell aggregation assay. In this assay, an equal number of HEK cells expressing a desired presynaptic cell adhesion molecule and a fluorescent protein are mixed with HEK cells expressing a post-synaptic cell adhesion molecule and another fluorescent protein. The interaction between the cell adhesion molecules forces the cells to aggregate, which can be...
07:03
An in vivo Assay to Test Blood Vessel Permeability
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Cited by 335 •
2013
We are presenting an in vivo assay to test blood vessel permeability. This assay is based on intravenous injection of a dye and subsequent visualization of its diffusion into interstitial spaces.
FAQs
Localized swelling is generated when antigen-responsive memory T cells in the transferred peripheral blood mononuclear cell population become activated in the mouse footpad. They release inflammatory cytokines, and the resulting local inflammation produces the delayed-type hypersensitivity signal. Thus, measured swelling reflects a cellular immune response rather than simply the presence of transferred cells.
Antigen-only and control-cell conditions establish how much swelling requires both the test antigen and immune-cell activity. Comparing the test injection with antigen alone helps identify antigen-dependent inflammation, while comparison with control cells helps distinguish responses associated with the transferred immune population. These controls make swelling an interpretable relative indicator of cellular immunity.
Immune-regulating conditions can reduce or alter the swelling produced by antigen-responsive cells. This makes the assay useful for examining suppression, including regulatory T-cell activity, rather than measuring response strength alone. A stronger signal indicates more pronounced localized cellular reactivity under the tested condition, whereas reduced swelling can indicate regulation of the transferred response.
Specificity is evaluated by pairing transferred immune cells with a defined test antigen and comparing the resulting swelling with responses to antigen alone or control-cell conditions. A difference attributable to the combination of immune cells and the test antigen supports antigen-specific cellular reactivity. This comparison helps distinguish memory responses from nonspecific local inflammation.
The workflow begins with human or animal peripheral blood mononuclear cells, which are combined with the test antigen and injected into the footpad of an immunodeficient mouse. Parallel injections can contain antigen alone, control cells, or an immune-regulating condition. Investigators then compare localized footpad swelling across these conditions to estimate response strength and regulation.
The assay can evaluate peripheral blood mononuclear cells from either human or animal sources. This flexibility allows investigators to examine cellular responses from different experimental subjects while using the same antigen-plus-cell transfer format. The resulting footpad swelling provides a shared in vivo readout for comparing antigen-specific memory or immune regulation across those samples.
In infection research, the assay can assess pathogen-specific immune memory by testing transferred cells with a relevant antigen. It can also help evaluate vaccine responses, because swelling reports the strength of antigen-directed cellular immunity. Including control-cell, antigen-only, or immune-regulating conditions adds information about whether the response is strong, antigen-dependent, or suppressed.