Cytotoxic T lymphocyte activation assay
AssigneeThe United States of America as represented by the Department of Health and Human Services
InventorMichail V. Sitkovsky
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Abstract
A method is provided for the quantitative study of cytotoxic T-lymphocyte activation by measuring secreted granule-associated BLT esterase activity after incubating the cytotoxic T-lymphocytes with activating stimuli. This method can be used to screen inhibiting (drugs) or activating agents (monoclonal antibodies) (at selected sub-optimal levels of activation of cytotoxic T-lymphocytes.
Claims11
A method for directly assaying cytotoxic T lymphocyte activation comprising the steps of:
- 1) incubating cytotoxic T lymphocytes with one or more lymphocyte activating stimuli selected from the group consisting of immobilized anti-TcR monoclonal antibody, an immunocomplex of anti-TcR monoclonal antibody and immobilized rabbit anti-mouse antibody, a mixture of beta-phorbol-12-myristate-13 acetate and ionophore A23187, and immobilized concanavalin A, said activating stimuli inducing the exocytosis of cytotoxic T lymphocyte granules and resulting in the secretion of BLT-esterase;
- and
- 2) quantitatively or qualitatively measuring the secretion of said BLT-esterase resulting from the activation of cytotoxic T lymphocyte by the lymphocyte activating stimuli.
A method for screening monoclonal antibodes or pharmacological agents for cytotoxic T lymphocyte activation comprising the steps of:
- 1) incubating cytotoxic T lymphocytes with one or more monoclonal antibodies or pharmacological agents;
- 2) quantitatively or qualitatively measuring the exocytosis of cytotoxic T lymphocyte granules by the resulting secretion of BLT-esterase, said secretion, if any, being induced by cytotoxic T lymphocyte activating monoclonal antibodies or pharmacological agents.
A method for screening monoclonal antibodies or pharmacological agents for use in inhibiting activation of cytotoxic T lymphocytes comprising the steps of:
- 1) incubating cytotoxic T lymphocytes with:
- a) one or more monoclonal antibodies or pharmacological agents, and
- b) one or more lymphocyte activating stimuli selected from the group consisting of immobilized anti-TcR monoclonal antibody, an immunocomplex of anti-TcR monoclonal antibody and immobilized rabbit anti-mouse antibody, a mixture of beta-phorbol-12-myristate-13 acetate and ionophore A23187, and immobilized concanavalin A, said activating stimuli inducing the exocytosis of cytotoxic T lymphocyte granules resulting in the secretion of BLT-esterase;
- and
- 2) quantitatively or qualitatively measuring the exocytosis of cytotoxic T lymphocyte granules by the resulting secretion, if any, of BLT-esterase.
Dependencies are inferred from the claim text.