Immune Profiling Core

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ABSTRACT – Immune Profiling Core The Immune Profiling Core (IPC) will provide comprehensive technical assay support services for the hypothesis-driven research studies proposed in the two Projects comprising this U19 proposal. The Core derives its greatest overall strength from the relevant expertise, organization, procedures, and operational synergy of its components. Anchoring these efforts is the Duke Immune Profiling Core (DIPC), a Duke School of Medicine Shared Resource that has been in existence since 1988. Largely focused on flow cytometry-based platforms for human immune profiling, the Core has actively participated in external proficiency testing and quality assurance programs for over 25 years. Assay testing is performed in a Good Clinical Laboratory (GCLP)-compliant environment according to highly standardized and validated Standard Operating Procedures (SOPs). The Core is joined in its efforts by a support team of highly experienced laboratory professionals from the Kirk and Knechtle research Cores who have recently re-located to Duke from Emory University. These individuals have extensive expertise in all aspects of transplantation research, both in human and nonhuman primate (NHP) studies. The key services the IPC will provide to Projects 1 and 2 include: 1) state-of-the-art specimen processing and biobanking, 2) analytical polychromatic flow cytometry (PFC) phenotyping, and intracellular cytokine staining (ICCS), 3) virologic testing, and 4) comprehensive histology and immunohistochemistry support. The IPC is well positioned to provide PFC services to the projects under this proposal; PFC having been the technical mainstay of the research endeavors of the DIPC. The present core facility houses a 4-laser BD LSRII, 5-laser BD LSRFortessa, 5-laser BD LSR-X20 and, most recently, a 5-laser BD LSR-X50. Of these digital instruments, the first 3 are capable of up to 18-24 parameter single cell analyses, while the X50 is capable of supporting 50-parameter flow cytometry. To provide comprehensive immunologic support, we have implemented several highly developed components for the IPC. A specimen processing and biobanking service that began at Emory under Dr. Kirk has been re-established as the Substrate Services and Research Support Service (SSCRSS) at Duke following his arrival as Chairman of the Department of Surgery. Through the work of Dr. Kirk's laboratory, we have also established a viral testing facility that will allow the IPC to monitor three of the most clinically-relevant latent viruses in NHPs: Rhesus Cytomegalovirus (rhCMV), Rhesus Lymphocryptovirus (rhLCV) (rhesus homology of EBV), and Simian Virus 40 (SV40). The IPC will provide routine histology and immunohistochemistry assays through our immunohistochemistry service laboratory, which will allow each project to examine the effects of the experimental manipulations on key host tissue. Special emphasis will be placed on examining effects on germinal center formation following the various induction regimens. The juxtaposition of human and NHP expertise represents a particular strength of this IPC consortium, further facilitating the highly translational aspects of the proposed studies.