The Immune Monitoring Core will be comprised of technologies and services offered by two service laboratories within the Duke Human Vaccine Institute (DHVI): the Flow Cytometry Facility and the Immune Reconstitution Facility. The RadCCORE Immune Monitoring Core (Core C) will routinely provide investigators with high quality, state-of-the-art flow cytometry, protein array, and gene expression assay support for their basic and applied research efforts. In addition to providing these services, the Core will be instrumental in developing new methodologies and making them available to RadCCORE Investigators. Administrative oversight for the Core will institute a charge-back mechanism to remain revenue neutral.

 

 

The three goals of the Immune Monitoring Core are:

1) Provide state-of-the-art, multi-color, fluorescence activated cell sorting and analytical support for basic or translational research conducted by Duke RadCCORE investigators. This includes high-speed sorting and multi-color analysis of cellular subsets on our 8-color BD FACSVantage SE, 7-color BD LSRII, 4-color FACSCalibur and 13-color BD FACSAria. Assays are available for surface and intracellular marker analysis of mouse, rat, rhesus, baboon and human cells.

 

2) Provide targeted multiplex protein array profiling of biological samples, such as tissue culture supernatant, serum/plasma, and lung lavage fluid, using our BioPlex bead array reader (BioRad). This instrumentation and Luminex bead technology has the capability of simultaneously quantifying up to 100 distinct analytes (e.g. cytokines, chemokines, hormones, signaling transduction proteins) in a single 50ul sample. Reagents are available for analysis of mouse, rat, and human samples.

 

3) Provide quantitative real-time-PCR multiplex gene expression analysis to investigators on our real-time Optical I-Cycler (BioRad). Specific assays include T cell receptor excision circle quantification to monitor thymopoiesis in mice, humans and monkeys (qPCR), and simultaneous quantification of up to four target mRNA species in a single sample (qRT-PCR). In consultation with the core RadCCORE investigators will be able to design, QC and implement a real-time quantitative PCR assay for their particular gene(s) of interest using either SYBR green or primer/probe TaqMan detection approaches.