The Flow Cytometry Core (FCC) will provide three primary services to support the scientific aims of the four proposed projects;population purification using cell sorting, cytokine identification and quantification using bead based cytokine arrays (Luminex system), and multispectral imaging flow cytometry (MIFC) using the ImageStream system. The first of these services, cell sorting, will be accomplished using a BD DiVa FACSVantage high speed sorter or the recently acquired i-Cyt Reflection high speed sorter. Both of these instruments are full equipped to sort the populations of interest described in the aims of Projects 1, 2, and 3. Evaluation of cytokine profiles of serum/plasma, tissue homogenates and culture supernatants will be accomplished! using the Luminex bead based multiplexing technology. Screening of up to 32 analytes will be available as well as customized panels of analytes of specific interest to accommodate the aims of all four projects. The FCC will provide sample preparation, data acquisition and data analysis for these assays. Thirdly, with the use of the ImageStream imaging flow cytometer, multispectral images as well as statistically robust data will be available to evaluate and confirm protein colocalizations, cytokine production, and cellular activation, as well as provide additional morphological information on cellular populations from the lung, spleen, liver and lymph node. In addition, the FCC has available instrumentation should the need arise to support polychromatic flow cytometry in excess of the capability of the Project leaders'own instrumentation. The FCC will also provide consultative services to the Project leaders in terms of fluorochrome selection, sample preparations, data analysis, and data interpretation. Likewise, the FCC Core Director will coordinate with the Director of the Histology Core (Core B) to insure comprehensive and complimentary services for all four Projects for the evaluation of immune interactions in the process of viral clearance and tissue injury. Flow Cytometry Core will provide the tools to a)isolate specific subsets of cells to further investigate their role in this process, b) identify and quantify the milieu of secreted cytokines/chemokines driving the response, c) capture morphological and fluorescent images of specific cell types identified as key components.

Public Health Relevance

Availability of the appropriate Flow Cytometric technologies is critical to the success of the proposed projects which attempt to dissect the role of the host's innate and adaptive immune responses in viral clearance and tissue destruction from inflammatory responses.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Program--Cooperative Agreements (U19)
Project #
5U19AI083024-05
Application #
8474691
Study Section
Special Emphasis Panel (ZAI1-BDP-I)
Project Start
Project End
Budget Start
2013-06-01
Budget End
2014-05-31
Support Year
5
Fiscal Year
2013
Total Cost
$93,747
Indirect Cost
$24,911
Name
University of Virginia
Department
Type
DUNS #
065391526
City
Charlottesville
State
VA
Country
United States
Zip Code
22904
Teoh, Jeffrey J; Gamache, Awndre E; Gillespie, Alyssa L et al. (2016) Acute Virus Control Mediated by Licensed NK Cells Sets Primary CD8+ T Cell Dependence on CD27 Costimulation. J Immunol 197:4360-4370
Jiang, Li; Yao, Shuyu; Huang, Su et al. (2016) Type I IFN signaling facilitates the development of IL-10-producing effector CD8(+) T cells during murine influenza virus infection. Eur J Immunol 46:2778-2788
DeBerge, Matthew P; Ely, Kenneth H; Wright, Peter F et al. (2015) Shedding of TNF receptor 2 by effector CD8⁺ T cells by ADAM17 is important for regulating TNF-α availability during influenza infection. J Leukoc Biol 98:423-34
Kim, Taeg S; Hanak, Mark; Trampont, Paul C et al. (2015) Stress-associated erythropoiesis initiation is regulated by type 1 conventional dendritic cells. J Clin Invest 125:3965-80
Krueger, Peter D; Kim, Taeg S; Sung, Sun-Sang J et al. (2015) Liver-resident CD103+ dendritic cells prime antiviral CD8+ T cells in situ. J Immunol 194:3213-22
Ramana, Chilakamarti V; DeBerge, Matthew P; Kumar, Aseem et al. (2015) Inflammatory impact of IFN-γ in CD8+ T cell-mediated lung injury is mediated by both Stat1-dependent and -independent pathways. Am J Physiol Lung Cell Mol Physiol 308:L650-7
Steinke, John W; Liu, Lixia; Turner, Ronald B et al. (2015) Immune surveillance by rhinovirus-specific circulating CD4+ and CD8+ T lymphocytes. PLoS One 10:e0115271
Moser, Emily K; Sun, Jie; Kim, Taeg S et al. (2015) IL-21R signaling suppresses IL-17+ gamma delta T cell responses and production of IL-17 related cytokines in the lung at steady state and after Influenza A virus infection. PLoS One 10:e0120169
Moser, Emily K; Hufford, Matthew M; Braciale, Thomas J (2014) Late engagement of CD86 after influenza virus clearance promotes recovery in a FoxP3+ regulatory T cell dependent manner. PLoS Pathog 10:e1004315
DeBerge, Matthew P; Ely, Kenneth H; Enelow, Richard I (2014) Soluble, but not transmembrane, TNF-α is required during influenza infection to limit the magnitude of immune responses and the extent of immunopathology. J Immunol 192:5839-51

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