Antigen-Specific Human T Cell Responses; Extremely Sensitive ELISA
We developed methods to investigate the effect of drugs on human antigen-specific T cell responses. Blood mononuclear cells or T lymphocytes from donors who had recently been vaccinated against cytomegalovirus (CMV) or other antigen were given a secondary challenge with the respective antigen or peptide epitope, or with irrelevant antigen.
Secondary response of CMV immunized cells to CMV peptide in vitro but not by irrelevant influenza peptide. Monoclonal antibody to CD3 (mAb CD3) is a positive nonspecific control.
Human IFNg ELISA sensitive to <1.3pg/mL
GPCR Bioassay, PTH Induction of cAMP
G protein-coupled receptors (GPCRs) are a large protein family of receptors that sense molecules outside the cell and activate inside signal transduction pathways and cellular responses. There are two principal pathways: cAMP and phosphatidylinositol.
Parathyroid hormone (PTH) regulates calcium ion homeostasis and induces cAMP in the osteosarcoma UMR-106 cell line. Cells are incubated with agonist for 15 minutes and cAMP production is measured by chemiluminescence.
Potency of a Test Compound: The cAMP accumulation is shown as percentage of a no stimulation control. The EC50 was calculated as 0.21nM by a 4P logistic fit.
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