This research will develop chemiluminescent-based system for the quantitation of lymphocyte subpopulstiions in whole blood. The technique will use an alkaline phosphate -based detection system with s chemiluminescent enzyme substrate to quantitate and identify humsn pripheral blood lymphocytes on instant film. When dephosphorylated by alkaline phosphatase, the chemiluminescent substrate 3-(2-adamantane)-4(3"-phosphoryloxy)phenyl-1,2-dioxetane(AMPPD) results in a glow of light that may be imaged on Polaroid instant film. Initially, cells that contain the enzyme will be detected by direct incubation with the colorimetric and chemiluminescent substrates.After optimization of the quantitation steps, cell surface labeling will be used with monoclonal antibodies to identify and quantitate lymphocyte subsets within a sample of whole blood. In Phase II of the project, Tropix, Inc., will correlate the data obtained on instant film with subtyping experiments performed with fluorescent monoclonal antibodies on the FACS in order to develop a clinical assay for lymphocyte subtyping.Awardee's statement of the potential commercial applications of the research:The goal of this work is to develop a rapid, sensitive method for identifying and quantitating blood cell types without the need for expensive instrumentation or technical expertise.National Institute of Allergy and Infectious Diseases (NIAID)
