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AssigneeOncolmmunin, Inc.
InventorsBeverly Packard, Martin J. Brown, Mark Feinberg, Luzheng Liu, Guido Silvestri, Ann Chahroudi, Akira Komoriya
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This invention provides a non-radioactive assay to monitor and quantify the target-cell killing activities mediated by cytotoxic T lymphocytes (CTLs). This assay is predicated on the discovery that apoptosis pathway activation and, in particular, caspase activity, provides a measure of cytotoxic effector cell activity. In one embodiment, measurement of CTL-induced caspase activation in target cells is achieved through detection of the specific cleavage of fluorogenic caspase substrates. This assay reliably detects antigen-specific CTL killing of target cells, and provides a more sensitive, more informative and safer alternative to the standard 51Cr-release assay most often used to quantify CTL responses. The assay can be used to study CTL-mediated killing of primary host target cells of different cell lineages, and enables the study of antigen-specific cellular immune responses in real time at the single-cell level. As such, the assay can provide a valuable tool for studies of infectious disease pathogenesis and development of new vaccines and immunotherapies.
A method of detecting cell-mediated cytotoxic activity, said method comprising:
A method of detecting cell-mediated cytotoxic activity, said method comprising:
The method of claim 81, wherein said target cell is selected from the group consisting of a tumor cell, a neural cell, a muscle cell, a fibroblast, a connective tissue cell, a bone cell, a blood cell, a spinal fluid derived cell, a lymphatic fluid derived cell, and a cell obtained from the site of an inflammation.
A method of detecting cell-mediated cytotoxic activity, said method comprising:
A method of detecting the presence of memory cytotoxic effector activity, said method comprising:
The method of claim 105, wherein said ligand comprises a subsequence of a polypeptide selected from the consisting of PARP, nuclear lamin, actin, PKC gamma, SREBP, U1-RNP, DNA-PK, G4-GDI, huntingtin, and HnRNP-C1/2, wherein said subsequence is of sufficient length to specifically bind to the substrate binding site of said activated caspase.
Dependencies are inferred from the claim text.