ORGANIZATION AND BUDGET OF THE CORE: The NEI Flow Cytometry Core has been operational for 26 months. It is equipped with two FACSCaliburs (Ages: 3 years and 12 years old) and one FACSAria (Age: 5 years old). The Core operates from 2 small, noncontiguous, storage rooms totaling 137 square feet. The FACSCalibur workspace totals 77 square feet of which the machines'footprint accounts for 60% of the space. The FACSAria workspace totals 60 square feet of which the machine itself and its accessories use 80% of the space. The Core is operated by a Core Manager devoting 100% effort to the Core and is supervised by a PI devoting 7% effort to the Core. The Core utilized 100% of its budget plus a $112,500 contribution from the NEI Office of Scientific Director for its routine operations and for purchase of a new FACSAria II that is replacing the 5-year old FACSAria I starting in the new fiscal year. OPERATION OF THE CORE: This year, over fifty individuals from thirteen different laboratories used the facility. These services and collaborative services were performed for 9 Principal Investigators (PIs) from 5 NEI labs (LI, LRCMB, N-NRL, OSD and URN), plus 3 PIs from 2 other institutes at NIH (NIAMS, and NICHD). Over 16,000 samples were analyzed. Seven users received training on data acquisition using the FACSCaliburs. Twenty-one users required cell sorting using the FACSAria. This year 260 sorts were performed, an 80% increase from the previous year. Among the techniques now in use within the core there are methods for phenotyping live cells, detecting gene expression, monitoring membrane and DNA content changes due to apoptosis, measurement of intracellular proteins and quantification of soluble proteins, The work involving human tissues is limited to sorting of peripheral blood mononuclear cells to study their cytokine production, genotype and RNA expression. No tissues were stored. TRAINING: Five courses were provided by the core to the NEI community. These courses included: Introduction to Flow Cytometry, Basics of Flow Cytometry, and FACSCalibur Operation. The Core Manager completed 48 hours of continuing education in flow cytometry and 12 hours of training in management of Core facilities. Publications generated by this research: 1. Jun Tang, Ru Zhou, Dror Luger, Wei Zhu, Phyllis B. Silver, Rafael S. Grajewski, Shao-Bo Su, Chi-Chao Chan, Luciano Adorini, and Rachel R. Caspi. Calcitriol Suppresses Antiretinal Autoimmunity through Inhibitory Effects on the Th17 Effector Response. The Journal of Immunology, 2009, 182, 4624 -4632 2. Ding X, Patel M, Shen D, Herzlich AA, Cao X, Villasmil R, Klupsch K, Tuo J, Downward J, Chan CC. Enhanced HtrA2/Omi Expression in Oxidative Injury to Retinal Pigment Epithelial Cells and Murine Models of Neurodegeneration. Invest. Ophthalmol. Vis. Sci. 2009 May 14.

National Institute of Health (NIH)
National Eye Institute (NEI)
Scientific Cores Intramural Research (ZIC)
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St Leger, Anthony J; Desai, Jigar V; Drummond, Rebecca A et al. (2017) An Ocular Commensal Protects against Corneal Infection by Driving an Interleukin-17 Response from Mucosal ?? T Cells. Immunity 47:148-158.e5
Zárate-Bladés, Carlos R; Horai, Reiko; Mattapallil, Mary J et al. (2017) Gut microbiota as a source of a surrogate antigen that triggers autoimmunity in an immune privileged site. Gut Microbes 8:59-66
Dillenburg-Pilla, Patricia; Zárate-Bladés, Carlos R; Silver, Phyllis B et al. (2016) Preparation of Protein-containing Extracts from Microbiota-rich Intestinal Contents. Bio Protoc 6:
Kielczewski, Jennifer L; Horai, Reiko; Jittayasothorn, Yingyos et al. (2016) Tertiary Lymphoid Tissue Forms in Retinas of Mice with Spontaneous Autoimmune Uveitis and Has Consequences on Visual Function. J Immunol 196:1013-25
Zárate-Bladés, Carlos R; Horai, Reiko; Caspi, Rachel R (2016) Regulation of Autoimmunity by the Microbiome. DNA Cell Biol 35:455-8
Ma, Wenxin; Cojocaru, Radu; Gotoh, Norimoto et al. (2013) Gene expression changes in aging retinal microglia: relationship to microglial support functions and regulation of activation. Neurobiol Aging 34:2310-21
Horai, Reiko; Silver, Phyllis B; Chen, Jun et al. (2013) Breakdown of immune privilege and spontaneous autoimmunity in mice expressing a transgenic T cell receptor specific for a retinal autoantigen. J Autoimmun 44:21-33
Chong, Wai Po; Ling, Man To; Liu, Yinping et al. (2013) Essential role of NK cells in IgG therapy for experimental autoimmune encephalomyelitis. PLoS One 8:e60862
Wang, Yujuan; Shen, Defen; Wang, Vinson M et al. (2012) Enhanced apoptosis in retinal pigment epithelium under inflammatory stimuli and oxidative stress. Apoptosis 17:1144-55
Shi, Guangpu; Lovaas, Jenna D; Tan, Cuiyan et al. (2012) Cell-cell interaction with APC, not IL-23, is required for naive CD4 cells to acquire pathogenicity during Th17 lineage commitment. J Immunol 189:1220-7

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