Imaging Synapse Formation during Thymocyte Selection Inability of CD3ζ to Form a Stable Central Accumulation during Negative Selection

AbstractTCR signaling can result in cell fates ranging from activation to tolerance to apoptosis. Organization of molecules in an “immunological synapse” between mature T cells and APCs correlates with the strength of TCR signaling. To investigate synapse formation during thymic selection, we have established a reaggregate system in which molecular recruitment of GFP fusion proteins to thymocyte:stromal cell interfaces can be visualized in real time. We demonstrate that negative selection is associated with efficient conjugate formation and rapid recruitment of p56lck and CD3ζ to an immunological synapse. Interestingly, CD3ζ-GFP does not accumulate at the center of the synapse, as in mature T cells, but at the periphery across a wide range of ligand densities. This implicates differences in synapse geometry in initiation of alternate signals downstream of the TCR

T-Cell activation: a queuing theory analysis at low agonist density

We analyze a simple linear triggering model of the T-cell receptor (TCR) within the framework of queuing theory, in which TCRs enter the queue upon full activation and exit by downregulation. We fit our model to four experimentally characterized threshold activation criteria and analyze their specificity and sensitivity: the initial calcium spike, cytotoxicity, immunological synapse formation, and cytokine secretion. Specificity characteristics improve as the time window for detection increases, saturating for time periods on the timescale of downregulation; thus, the calcium spike (30 s) has low specificity but a sensitivity to single-peptide MHC ligands, while the cytokine threshold (1 h) can distinguish ligands with a 30% variation in the complex lifetime. However, a robustness analysis shows that these properties are degraded when the queue parameters are subject to variation—for example, under stochasticity in the ligand number in the cell-cell interface and population variation in the cellular threshold. A time integration of the queue over a period of hours is shown to be able to control parameter noise efficiently for realistic parameter values when integrated over sufficiently long time periods (hours), the discrimination characteristics being determined by the TCR signal cascade kinetics (a kinetic proofreading scheme). Therefore, through a combination of thresholds and signal integration, a T cell can be responsive to low ligand density and specific to agonist quality. We suggest that multiple threshold mechanisms are employed to establish the conditions for efficient signal integration, i.e., coordinate the formation of a stable contact interface

Ecological assessments at DOE hazardous waste sites: Current procedures and problems

Major actions at US Department of Energy (DOE) hazardous waste sites require CERCLA compliance that meets NEPA considerations. Although NEPA compliance includes ecological considerations, neither the Council on Environmental Quality (CEQ) nor the DOE provide detailed guidance for conducting ecological assessments under NEPA. However, the identification of the form and magnitude of potential ecological impacts associated with a proposed action is directly dependent on the quality of the baseline data available for a particular site. Using the Surplus Facilities Management Program Weldon Spring site as an example, we discuss the collection of baseline ecological data for the site. This site is surrounded by approximately 17,000 acres of wildlife area. Available wildlife data consisted of qualitative, county-level species lists, and vegetation data was in the form of a regional qualitative narrative. Detailed site-specific occurrence data for listed species and high quality natural communities was provided by the Missouri Department of Conservation Heritage data base. 30 refs., 1 fig., 1 tab

Land cover change and remote sensing: Examples of quantifying spatiotemporal dynamics in tropical forests

Research on human impacts or natural processes that operate over broad geographic areas must explicitly address issues of scale and spatial heterogeneity. While the tropical forests of Southeast Asia and Mexico have been occupied and used to meet human needs for thousands of years, traditional forest management systems are currently being transformed by rapid and far-reaching demographic, political, economic, and environmental changes. The dynamics of population growth, migration into the remaining frontiers, and responses to national and international market forces result in a demand for land to produce food and fiber. These results illustrate some of the mechanisms that drive current land use changes, especially in the tropical forest frontiers. By linking the outcome of individual land use decisions and measures of landscape fragmentation and change, the aggregated results shows the hierarchy of temporal and spatial events that in summation result in global changes to the most complex and sensitive biome -- tropical forests. By quantifying the spatial and temporal patterns of tropical forest change, researchers can assist policy makers by showing how landscape systems in these tropical forests are controlled by physical, biological, social, and economic parameters