Cellular interactions between antigen (Ag)-specific T cells and Ag-presenting cells (APCs) can trigger either productive or aborted immune responses. The cellular outcomes depend on the state of the T cells, their previous exposure to Ag, and the APCs. This application addresses the central hypothesis that the molecular architecture of the membrane of T cells plays a decisive role in influencing Ag-sensitivity and cellular outcomes.
Aim 1 will use immuno-electron microscopy, FRET and Quantum Dot blinking to test the hypothesis that antigenic stimulation of naTve T cells results in long-term formation of TCR micro-clusters that facilitate responses to recall Ags.
Aim 2 will use multi-dimensional digital imaging and biochemical analysis to identify specific antigen-induced spatial-temporal molecular alterations in Immunological Synapses that trigger altered immune responses.
This Aim will test the hypothesis that disproportionate activation of Fyn and Lck in the Immunological Synapse can cause unresponsiveness and would address the functional significance of the Immunological Synapse.
Aim 3 will test more directly the roles of selective activation of Fyn or Lck on the structure of the synapse.
This Aim will use novel kinase-null T cellls and switchable mutated inactive tyrosine kinases Fyn and Lck that can be chemically rescued rapidly and reversibly (within 5 min) in T cells. Uncovering the mechanisms by which T cells can sense Antigen and how this recognition determines the cellular response is the long-term objective of all 5 projects in this Program. There is an extremely high synergy with all other projects.
Aim 1 synergizes with Project 1 that will study TCR clustering in the membrane of CDS cells.
Aim 2 synergizes with Projects 3, 4 and 5, which will use the specialized imaging tools in their own structure function studies of Sproutyl, TFII-I and CARD11 and their associations with the Immunological Synapse.
Aim 3 would synergize with Project 4 where all the tools for chemical rescue of Fyn and Lck will be developed. These studies that combine uniquely the use of several highly novel experimental approaches are very likely to generate significant better understanding of the integrated events that regulate immune responses to infectious pathogens. This knowledge may lead to the design of more effective vaccines, better immune surveillance protocols, and new immunosupressive drugs.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Program Projects (P01)
Project #
5P01AI072677-05
Application #
8381799
Study Section
Special Emphasis Panel (ZAI1-PA-I)
Project Start
Project End
2014-08-31
Budget Start
2012-09-01
Budget End
2013-08-31
Support Year
5
Fiscal Year
2012
Total Cost
$319,510
Indirect Cost
$96,147
Name
Johns Hopkins University
Department
Type
DUNS #
001910777
City
Baltimore
State
MD
Country
United States
Zip Code
21218
Bettencourt, Ian A; Powell, Jonathan D (2017) Targeting Metabolism as a Novel Therapeutic Approach to Autoimmunity, Inflammation, and Transplantation. J Immunol 198:999-1005
Kosmides, A K; Meyer, R A; Hickey, J W et al. (2017) Biomimetic biodegradable artificial antigen presenting cells synergize with PD-1 blockade to treat melanoma. Biomaterials 118:16-26
Tiper, Irina V; Temkin, Sarah M; Spiegel, Sarah et al. (2016) VEGF Potentiates GD3-Mediated Immunosuppression by Human Ovarian Cancer Cells. Clin Cancer Res 22:4249-58
Pollizzi, Kristen N; Sun, Im-Hong; Patel, Chirag H et al. (2016) Asymmetric inheritance of mTORC1 kinase activity during division dictates CD8(+) T cell differentiation. Nat Immunol 17:704-11
Schütz, Christian; Varela, Juan Carlos; Perica, Karlo et al. (2016) Antigen-specific T cell Redirectors: a nanoparticle based approach for redirecting T cells. Oncotarget 7:68503-68512
Pollizzi, Kristen N; Waickman, Adam T; Patel, Chirag H et al. (2015) Cellular size as a means of tracking mTOR activity and cell fate of CD4+ T cells upon antigen recognition. PLoS One 10:e0121710
Makowski, Stefanie L; Wang, Zhaoquan; Pomerantz, Joel L (2015) A protease-independent function for SPPL3 in NFAT activation. Mol Cell Biol 35:451-67
Shaikh, Saame Raza; Boyle, Sarah; Edidin, Michael (2015) A high fat diet containing saturated but not unsaturated fatty acids enhances T cell receptor clustering on the nanoscale. Prostaglandins Leukot Essent Fatty Acids 100:1-4
Pollizzi, Kristen N; Patel, Chirag H; Sun, Im-Hong et al. (2015) mTORC1 and mTORC2 selectively regulate CD8? T cell differentiation. J Clin Invest 125:2090-108
Perica, Karlo; Bieler, Joan Glick; Schütz, Christian et al. (2015) Enrichment and Expansion with Nanoscale Artificial Antigen Presenting Cells for Adoptive Immunotherapy. ACS Nano 9:6861-71

Showing the most recent 10 out of 34 publications