The Human Tissue Acquisition and Processing Core (Scientific Core B) coordinates the acquisition ofhuman tissues from which mast cells, basophils and B cells are obtained. Mast cells derived from tissuesand basophils and B cells from peripheral blood, after they have matured in their natural environments,best reflect the functional attributes of these cells in vivo. They lack the chromosomal aberrations andgene expression abnormalities of leukemic cells, and also exhibit key differences from those cells derivedin vitro from progenitors. For example, cord blood-derived mast cells express primarily FcvRllb, aninhibitory receptor, while those dispersed from skin and lung express only FcyRlla, an activating receptor.Fresh surgical lung and skin will be used as sources of mature human mast cells, peripheral blood forbasophils and B cells. The MCT cell is the predominant mast cell type in lung, the MCTc cell in skin. TheNational Diseases Research Interchange (www.ndriresource.org/), Cooperative Human TissueNetwork (http://www-chtn.ims.nci.nih.gov/) and Pathology Department at VCD will provide skin and lung;and the Virginia Blood Services (www.vablood.org/) (buffy coat cells) and individual donors willprovide a source of peripheral blood. Mast cells will be dispersed from lung and skin, purified with anti-KitmAb, separated into MCT and MCTc cells with anti-CD88 Ab as needed, placed into culture and distributedto the appropriate Projects. Basophils and B cells from buffy coats or peripheral blood will be purified bydensity-dependent sedimentation and negative selection, and distributed. Project 1 requires tissue-derivedmast cells and peripheral blood basophils to study FceRI and/or FcyRlla mediated desensitization, andfacilitates collaborations with Dr. Spiegel (P4) on the involvement of SphK and CerK and with Dr. Ryan(P3) on the involvement of Lyn in desensitization. Project 2 utilizes B cells from peripheral blood or buffycoats to extend the animal model of CD23-dependent regulation of IgE production to humans by assessingin vitro the effect of novel CD23 modulating agents on IgE production. Project 4 utilizes tissue-derivedmast cells to examine the effects of novel inhibitors of sphingosine and ceramide kinases on mediatorproduction by these cells after they are activated, and facilitates collaborations with Drs. Schwartz andKepley on the roles of these enzymes on human mast cell functions. Core B also will manage the OdysseyInfrared Imaging System, which will be used primarily for processing and analyzing Western blots of thesignal transduction molecules under study in Projects 1-4. Thus, Core B relates to each of the proposedProjects.
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