Project Summary Core F – Immune Function The Immune Function Core F provides broadly ranging support for immunological studies. Driven by investigator need, and a rapidly changing science environment, the Core currently offers multiplexed cytokine measurements, standard flow cytometric analysis as well as flow imaging, sorting of cell populations via flow cytometry and other methods, and ELISA, ELISPOT, and infrared imaging. The re-structuring of the Core in 2010 included a cooperative agreement with the Cancer Center Cytometry and Imaging Microscopy Facility to pool the equipment, harmonize services, and maintain formal access to CLIA-level flow cytometry. Drs. Scott Sieg and James Jacobberger have been working closely together as Co-directors and have used their complementary expertise to greatly enhance Core services. Dr. Sieg provides immunological expertise, functional assays, and biomarkers, especially in the context of HIV research; Dr. Jacobberger provides expertise on flow and image cytometry instrumentation, single-cell assays, and biomarker discovery, especially in the areas of cell cycle, apoptosis, and cell signaling. A major function of the Core is to collaborate with users to develop novel immunological assays. For example Core F, in collaboration with the Proteomics & Systems Biology Core G, is refining new flow cytometric assays for characterizing the phospho-proteome. This unique platform should open new avenues of research relevant to HIV pathogenesis, clinical evaluation, and treatment. A parallel effort is focused on developing multiple pathway signaling measurements on sub-populations of cells in unfractionated human samples (e.g., blood). The Core also interacts closely with the Clinical Core D for sample procurement. The Immune Function Core (F) is currently the one of the most heavily used of all the CWRU/UH CFAR cores and during this grant cycle the Core generated net revenues of $384,177. Program income has been steadily reinvested in the purchase of advanced instruments, such as multicolor flow cytometers, and reagents. For example in 2012, the Immune Function Core purchased a BD Fortessa – a next generation analytical instrument – with blue, green, red and violet lasers and filters optimized to measure fluorescent proteins. This instrument was subsequently upgraded to add a high throughput 96 well plate feed system thanks to a CFAR supplementary award. As part of the recruitment of Dr. Sékaly, the CFAR will purchase a BD Biosciences FACSAria with 4 or 5 lasers and 18-color capability. The FACSAria will be maintained under BSLII conditions to permit sorting of HIV-infected cells. This state-of-the-art instrument will bring sorting capacity in line with our analytical capacities and create further opportunities for Core-Core interactions since it will be used to prepare samples for sequencing and proteomic analyses by Cores E and G.. The specifically aims of the Core follow: To provide ...