|Andrew D. Luster, MD, PhD
|Shiv Pillai, MD, PhD
|Shannon Bromley, PhD
|James Moon, PhD
|Fred Preffer, PhD
|Aviv Regev, PhD
The major goal of the Immunology Core is to provide expertise in design, implementation, and analysis for immunological assays to members of the CSIBD by equipping the community with cost-effective access to crucial instruments, reagents, protocols, training, and education. The Core is centered on modern approaches to the study of the human immune system and their application to the study of IBD and related disorders.
The objectives of the Immunology Core are as follows:
- Provide CSIBD members with access to immunological resources that would otherwise be cost-prohibitive to individual researchers
- Offer CSIBD investigators access to cutting-edge immunological techniques that are at the forefront of immunology research
- Promote the development of junior investigators and foster collaborations by providing a connection point between investigators of varying research approaches
- Education and training services. A central goal of this core is to provide instruction for investigators in modern immunologic methods. The co-directors and the manager of the Immunology Core have extensive experience in modern immunologic techniques and a longstanding commitment to teaching. Three levels of supervision are offered: consultation, short-term supervision of investigators in a specific technique, and long-term guidance on project development. In each of the several areas of expertise described above, consultation is provided to CSIBD investigators with laboratories actively using these techniques. However, many investigator-Core interactions involve more intensive training to permit a CSIBD investigator to apply several of these techniques to a specific research project. This expectation follows from the complex multi-step nature of these techniques and the requirement for specific modifications for most applications. For these interactions, a CSIBD investigator, or a member of his or her laboratory, spends the necessary time to learn and modify the technique under the direct guidance of Core personnel.
A cornerstone of the educational activities of the Core is the MGH Immunology Seminar Series. This series takes place every week in a conference room adjacent to the Center for Immunology and Inflammatory Diseases (CIID; located at MGH East, Charlestown Navy Yard [CNY]-149) and is coordinated by the Core in conjunction with the Harvard Medical School Immunology series such that speakers give a seminar at Harvard Medical School on Wednesday and then give a seminar at MGH on Thursday.
- Routine immunology tools
- Flow cytometry for both basic flow cytometry using up to four colors (six parameters) as well more complex flow cytometric analysis using up to fourteen colors
- Cell sorting and leukocyte isolation using high-speed cell sorting
- Multiplex and ELISA cytokine assays including flow cytometer-based cytokine bead arrays and Luminex multiplex cytokine arrays
- Dendrimer-based in situ hybridization and immunofluorescence on both FFPE and frozen sections to identify and monitor activity of disease-associated clones at sites of inflammation. For in situ hybridization, the use of branched DNA probes provided by Affymetrix allows sensitive detection of RNA in tissues
- Specialized immunology services
- Next-generation sequencing to identify clonal expansions of both T and B cell subsets in the blood and in human tissues
- Single-cell cloning of activated B cells to match Ig H and L chain sequences and create disease-related human monoclonal antibodies to monitor cells and expression patterns during disease flares, as well as progression and response to therapies
- MHC class II tetramers to identify, purify and characterize disease-causing antigen-specific T cells. The extracellular domains of MHC class II alpha and beta chains can be enzymatically biotinylated using the BirA ligase and then tetramerized using fluorochrome-conjugated streptavidin.
- Antigen multimers to identify, purify and characterize disease-causing antigen-specific B cells; these are also created by enzymatic biotinylation of antigen and tetramerization using streptavidin
- Phosphoproteomics, CyTOF mass cytometry, and Luminex bead assays to analyze signaling pathways and protein expression in activated T and B cells in depth
- Novel pathway-based computational approaches and nanowire RNAi on primary T cells to aid the discovery of biologically relevant “nodes” that may represent targets of therapeutic relevance in disease-specific activated B and T cells. CSIBD members developed this technology, which has been successfully applied to identify molecular circuits that control the differentiation of naïve T cells.