Immune Reconstitution & Biomarker Analysis
Director: Gregory D. Sempowski, PhD

Shared Resource Summary:
The Immune Reconstitution and Biomarker Analysis Shared Resource is a service component of the Sempowski laboratory and the Regional Biocontainment Laboratory at Duke (GHRB). In addition to providing access to specialized instrumentation for Real-Time PCR, and Luminex-based multiplex ELISA, this Shared Resource assists investigators with the design and implementation of molecular, protein, and cellular assays to quantify immune reconstitution or immune responses in vitro and in vivo. A major accomplishment of this facility has been the development of a patented Real-Time PCR assay system to quantify mouse T-cell receptor excision circles. This assay has proven invaluable for monitoring thymopoiesis in mice.
Personnel/Contact Information:
Director: Gregory D. Sempowski, PhD Office Phone: (919) 684-4386 Lab Phone: (919) 684-3280 Fax: (919) 684-3282 Email: gsem@duke.edu	Manager: Jeffrey Hale, MS Office Phone: (919) 681-3299 Lab Phone: (919) 684-3280 Fax: (919) 684-3282 Email: hale0023@mc.duke.edu
Location: 1033 GHRB, DUMC Box 103020, Durham, NC 27710
Getting Started:
Please see the Common Tools sidebar to register to use facilities, schedule or request a job, and to retrieve data. A Hazard Assessment Form needs to be completed for each NEW project to determine the appropriate level of containment and the SOP needed to assure proper safety precautions are used when working with your samples. These forms must be emailed to the DHVI IR Facility Director or Manager at least 48 hours prior to starting a NEW project for review and approval. Hazard Assessment Form Current approved base rates are listed below for services. Discounts may apply to these rates based on membership in affiliated centers or programs. Please contact the DHVI Shared Resources Business Office at (919) 684-3349 or via email. DHVI Immune Reconstitution & Biomarker Analysis Rates
Facilities and Instruments:
BSL-1, -2 and -3 laboratory space for sample processing Abbott Cell Dyn 3700 Veterinary Hematology Analyzer BioRad Luminex Bead Array Readers (2) BioRad Optical iCycler for Real-time PCR
Services Provided:
Luminex Bead Array Assays Human Peripheral Blood Processing Cell Subset Isolation Quantitative PCR (qPCR; qRT-PCR) T Cell Receptor Excision Circle Assay (TREC) T Cell Receptor Spectratyping
Protocols and Documents:
Misc. Protocols Human PBMC Separation by Ficoll-Hypaque Human and Mouse cell isolation by Miltenyi MACS Beads One-step qRT-PCR (SYBR Green) One-step qRT-PCR (with PROBES) DNA and RNA isolation using Trizol Human Spectratyping/Immunoscope Mouse Spectratyping/Immunoscope TREC Assay Protocols/Documents Mouse / Human TREC Assay Mouse TREC assay unit in Current Protocols of Immunology Proteinase K Digestion for TREC Mouse TREC Assay System Patent Notice Material Transfer Agreement to receive components of the mouse TREC assay Luminex Bead Assays BioRad Quick Guide to Bioplex Cytokine Kits BioRad Bioplex Cytokine Kit Documentation BioRad DHVI Core Brochure (Special Pricing Available!) Safety SOP's Safety SOP for Sample Processing at BSL-1 and BSL-2 Safety SOP for sample processing at BSL-3
Affiliated Centers:
Duke Radiation Countermeasures Center Grant- AI-067798 (radccore.mc.duke.edu) Duke University Center for Translational Research Grant - AI-051445 Southeast Regional Center of Excellence for Biodefense and Emerging Infections (www.serceb.org) Duke Center for AIDS Research (www.cfar.duke.edu) Center for HIV-AIDS Vaccine Immunology (www.chavi.org)
Publications:
2006-2009 Lynch HE, Goldberg GL, Chidgey A, Van den Brink MR, Boyd R, Sempowski GD. Thymic involution and immune reconstitution. Trends Immunol. 2009 Jul. 30(7): 366-73. Epub 2009 Jun 18. Abstract Sarzotti-Kelsoe M, Win CM, Parrott RE, Cooney M, Moser BK, Roberts JL, Sempowski GD, Buckley RH. Thymic output, T cell diversity and T cell function in long-term human SCID chimeras. Blood. 2009 May 11. Epub ahead of print. Abstract Page, KM, Mendizibal, A, Prasad, VK, Martin, PL, Parikh, S, Wood, S, Sempowski, GD, Szabolcs, P, Kurtzberg, J. 2008. Post-transplant autoimmune hemolytic anemia and other autoimmune cytopenias are increased in very young infants undergoing unrelated donor unbilical cord blood transplantation. Biology of Blood and Marrow Transplantation. IN PRESS. Gruver, AL and Sempowski GD. 2008. Cytokine, Leptin and Stress-induced Thymic Atrophy. J. Leukocyte Biology. Abstract Howard, BH, Sempowski, GD, Scearce, RM, Liao, HX, Lee, DM, Lam, GK, Chen, H, Fadden, P, Haystead, T, and Haynes, BF. 2008. Murine CD7 Shares Antigenic Cross-reactivity with HSP-60. Hybridoma. 27:81-89. Abstract Henry, SC, Daniell, X, Indaram, M, Whitesides, JF, Sempowski, GD, Howell, D, Oliver, T, Taylor, GA. 2007. Impaired macrophage function underscores susceptibility to Salmonella in mice lacking Irgm1 (LRG-47). J Immunol. 179:6963-6972. Abstract Hudson, LL, Markert, ML, Devlin, BH, Haynes,BF and Sempowski, GD. 2007. Human T Cell Reconstitution in DiGeorge Syndrome and HIV-1 Infection. Seminar in Immunology. 19:297-309. Abstract Nordone, SK, Ignacio, GA, Su, L, Sempowski, GD, Golenbeck, DT, Li, L, Dean, GA. 2007. Failure of TLR4-driven NF-kappa B activation to stimulate virus replication in models of HIV type 1 activation. ARHRV. 23:1387-95. Abstract Gruver, AL, Hudson, LL and Sempowski, GD. 2007. Immunosenescence of Aging. J. Pathology. 211:144-156. Abstract Li FQ, Sempowski GD, McKenna SE, Laskowitz DT, Colton CA, Vitek MP. Apolipoprotein E-derived peptides ameliorate clinical disability and inflammatory infiltrates into the spinal cord in a murine model of multiple sclerosis. 2006. J Pharmacol Exp Ther. 2006 318:956-965. Abstract Pubmed Archive