We are requesting funds for purchase of a new CyTOF 2 mass cytometer, manufactured by DVS Sciences, Inc. to complement the existing flow cytometry instruments in the Flow Cytometry Core Facility at the La Jolla Institute for Allergy and Immunology (LJI). If awarded, this novel technology will be available to scientists at LJI and to serve as a regional resource to the San Diego research community. Mass cytometry is one of the newest and most powerful tools available to understand complex biological processes that cannot be attained with conventional flow cytometry. Mass cytometry technology is premised on the use of elements, or stable isotopes, as tags instead of fluorophores, with measurement of the tags using an Inductively Coupled Plasma Mass Spectrometer (ICP-MS). The advantage lies in the large number of available elements and stable isotopes (potentially greater than 100), the high resolution of the mass spectrometer between detection channels, and the large dynamic range of detection of the ICP-MS. These benefits, and others, result in the ability to perform high-dimensional multi-parameter assays in single cells without the need for compensation. High-dimensional cytometry can be used to: (i) identify new subsets of cells in parallel to known subset of cells, (ii) to identify multiple cell types using one complete panel ina single sample, (iii) provide a comprehensive analysis for phenotype and function of cell subsets by chemokine expression, activation status, cytokine production, proliferation and signaling events, (iv) provide more information on limited clinical samples and rare cell subsets, (v) to evaluate alterations in complex differentiation pathways, (vi) to simultaneously evaluate complex alterations in intracellular signaling pathways in cells. The research projects detailed in this application are led by 9 major and minor principal investigators who are funded from several NIH grants. External collaborators and investigators in the San Diego area will also be offered access to, and assistance with, this technology. If funded and placed into service, a CyTOF will undoubtedly facilitate even greater progress on current and future NIH-supported projects. This novel technology will directly impact projects to understand more about the cells of the immune system and their discrete functions in diseases as wide-ranging as asthma, atherosclerosis, inflammatory bowel disease, sickle-cell disease and viral infections.
|Winkels, Holger; Ehinger, Erik; Vassallo, Melanie et al. (2018) Atlas of the Immune Cell Repertoire in Mouse Atherosclerosis Defined by Single-Cell RNA-Sequencing and Mass Cytometry. Circ Res 122:1675-1688|
|Burel, Julie G; Lindestam Arlehamn, Cecilia S; Khan, Nabeela et al. (2018) Transcriptomic Analysis of CD4+ T Cells Reveals Novel Immune Signatures of Latent Tuberculosis. J Immunol 200:3283-3290|
|Tyler, Christopher J; Pérez-Jeldres, Tamara; Ehinger, Erik et al. (2018) Implementation of Mass Cytometry as a Tool for Mechanism of Action Studies in Inflammatory Bowel Disease. Inflamm Bowel Dis 24:2366-2376|
|Tian, Yuan; Babor, Mariana; Lane, Jerome et al. (2017) Unique phenotypes and clonal expansions of human CD4 effector memory T cells re-expressing CD45RA. Nat Commun 8:1473|
|Thomas, Graham D; Hamers, Anouk A J; Nakao, Catherine et al. (2017) Human Blood Monocyte Subsets: A New Gating Strategy Defined Using Cell Surface Markers Identified by Mass Cytometry. Arterioscler Thromb Vasc Biol 37:1548-1558|