The UTSW-SLE-CORT Flow Cytometry Core will operate within the existing infrastructure of theRheumatology Division, the Harold C. Simmons Arthritis Research Center and the Center for Immunology tosupport the research activities of Project investigators. The individual research programs proposed in theUTSW-SLE-CORT have common requirements for reagents and services. The Core will consist of a Directorand a Research Technician. The Core will utilize the Director's laboratory for preparation of samples and forexperiments. The Flow Cytometry Core will employ flow cytometry instruments that are currently available atUT Southwestern Medical Center. The proposed activities of the Core will be to provide custom reagents andservices that would not otherwise be available for the individual laboratories of the UTSW-SLE-CORT.
The specific aims of the Flow Cytometry Core are:(1) to develop standardized protocols for flow cytometricassays; (2) to generate fluorochrome-labeled antibodies and provide quality control for the flow cytometricassays; and (3) to establish a data archiving system for flow cytometry data. In the first aim the Core willdevelop antibody panels for the identification of murine (Project 1) and human (Projects 2, 3 and 4) SLAMfamily members, activation markers and intracellular molecules for B, T, NK, dendritic cell (DC) and myeloidcell subsets. We will work with project investigators to develop and optimize protocols for cell preparationand storage, kinetics of cell responses for phospho-protein assays, and polychromatic immunophenotypingassays. Of import, the same reagents and protocols can be utilized for the identification of many murine andhuman intracellular molecules including phospho-proteins. The goal of this aim is to develop and implementprotocols that will result in the generation of maximum information from limited murine lupus and human SLEpatient samples. In the second aim, the Core will generate appropriately labeled antibodies with novelfluorochromes for complex polychromatic flow cytometric assays. The flow cytometry core will provideroutine quality control checks of the antibodies, protocols and assays for project investigators of the UTSW-SLE-CORT. In the third aim, the Core will assist project investigators with data analysis and establish amechanism to archive unprocessed data. Another objective of this aim is to facilitate communicationconcerning optimized protocols and potentially important new observations between project investigators.
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