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Felix Scheuplein Deanna J. Lamont Matthew E. Poynter Jonathan E. Boyson David Serreze Lennart K. A. Lundblad Robert Mashal Robert Schaub 《PloS one》2015,10(10)
Invariant Natural Killer T (iNKT) cells are a T cell subset expressing an invariant T Cell Receptor (TCR) that recognizes glycolipid antigens rather than peptides. The cells have both innate-like rapid cytokine release, and adaptive-like thymic positive selection. iNKT cell activation has been implicated in the pathogenesis of allergic asthma and inflammatory diseases, while reduced iNKT cell activation promotes infectious disease, cancer and certain autoimmune diseases such as Type 1 diabetes (T1D). Therapeutic means to reduce or deplete iNKT cells could treat inflammatory diseases, while approaches to promote their activation may have potential in certain infectious diseases, cancer or autoimmunity. Thus, we developed invariant TCR-specific monoclonal antibodies to better understand the role of iNKT cells in disease. We report here the first monoclonal antibodies specific for the mouse invariant TCR that by modifying the Fc construct can specifically deplete or activate iNKT cells in vivo in otherwise fully immuno-competent animals. We have used both the depleting and activating version of the antibody in the NOD model of T1D. As demonstrated previously using genetically iNKT cell deficient NOD mice, and in studies of glycolipid antigen activated iNKT cells in standard NOD mice, we found that antibody mediated depletion or activation of iNKT cells respectively accelerated and retarded T1D onset. In BALB/c mice, ovalbumin (OVA) mediated airway hyper-reactivity (AHR) was abrogated with iNKT cell depletion prior to OVA sensitization, confirming studies in knockout mice. Depletion of iNKT cells after sensitization had no effect on AHR in the conducting airways but did reduce AHR in the lung periphery. This result raises caution in the interpretation of studies that use animals that are genetically iNKT cell deficient from birth. These activating and depleting antibodies provide a novel tool to assess the therapeutic potential of iNKT cell manipulation. 相似文献
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† Robert W. Neumar Scott M. Hagle †Donald J. DeGracia Gary S. Krause †Blaine C. White 《Journal of neurochemistry》1996,66(1):421-424
Abstract: Proteolytic degradation of numerous calpain substrates, including cytoskeletal and regulatory proteins, has been observed during brain ischemia and reperfusion. In addition, calpain inhibitors have been shown to decrease degradation of these proteins and decrease postischemic neuronal death. Although these observations support the inference of a role for μ-calpain in the pathophysiology of ischemic neuronal injury, the evidence is indirect. A direct indicator of μ-calpain proteolytic activity is autolysis of its 80-kDa catalytic subunit, and therefore we examined the μ-calpain catalytic subunit for evidence of autolysis during cerebral ischemia. Rabbit brain homogenates obtained after 0, 5, 10, and 20 min of cardiac arrest were electrophoresed and immunoblotted with a monoclonal antibody specific to the μ-calpain catalytic subunit. In nonischemic brain homogenates the antibody identified an 80-kDa band, which migrated identically with purified μ-calpain, and faint 78- and 76-kDa bands, which represent autolyzed forms of the 80-kDa subunit. The average density of the 80-kDa band decreased by 25 ± 4 ( p = 0.008) and 28 ± 9% ( p = 0.004) after 10 and 20 min of cardiac arrest, respectively, whereas the average density of the 78-kDa band increased by 111 ± 50% ( p = 0.02) after 20 min of cardiac arrest. No significant change in the density of the 76-kDa band was detected. These results provide direct evidence for autolysis of brain μ-calpain during cerebral ischemia. Further work is needed to characterize the extent, duration, and localization of μ-calpain activity during brain ischemia and reperfusion as well as its role in the causal pathway of postischemic neuronal injury. 相似文献
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W. O. van der Knaap Jacqueline F. N. van Leeuwen Tomasz Goslar Robert Krisai Willy Tinner 《Vegetation History and Archaeobotany》2012,21(1):37-60
Three mires and a small lake in the Swiss and Austrian Alps were studied palynologically at high resolution, covering the
last 1,000, 400, 50 and 1,200 years, respectively. Methodological lessons include: (1) Sub-decadal resolution in upper, little-decomposed
peat layers reveals recurrent marked fluctuations in both percentages and influx of regional tree-pollen types, reflecting
variations in pollen production rather than in plant-population sizes. (2) Intermittent, single-spectrum pollen maxima in
samples of sub-decadal resolution indicate pollen transport in clumps. This type of pollen transport may remain unrecognized
in sections with lower sampling resolution, which may then lead to inappropriate interpretation in terms of plant-population
sizes. (3) The detection of short-lived phases of human impact in decomposed peat requires sampling intervals as close as
0.2 cm. (4) PAR (pollen influx) may reflect vegetation dynamics more faithfully than percentages. Reliable PAR, however, is
difficult to achieve in Alpine mires due to past human impact on peat growth, even when complex depth–age modelling techniques
are used. Critical comparison of PAR with percentages is therefore essential. (5) Careful consideration of spatial scales
in pollen signals (local–regional and subdivisions) is essential for a realistic palaeo-ecological interpretation. Results
in terms of past human impact on vegetation are summarized as follows: (1) Trends in pollen types reflecting regional human
action are in general agreement with earlier findings for the western Swiss Alps, allowing for regional differences. (2) All
mires in the Alps investigated here and in an earlier study experienced human impact during the last millennium. The studied
small lake, lying in sub-alpine pasture, records forest dynamics at a lower elevation since a.d. 800. 相似文献
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A number of Arabidopsis thaliana mutants with abnormal rootmorphology have been isolated from a population of Agrobacterium-mediated seed transformants. Five of these mutants identifiedby their abnormal root morphology have been genetically andmorphologically characterised. Two mutants, 5905 and 1767 havedefective cellular elongation, mutant 7203 lacks an endodermalcell layer and mutants 4792 and 7133 exhibit abnormal radialcellular expansion. It is apparent from detailed analysis thatabnormal root morphology is accompanied by abnormal hypocotylphenotype. Genetic analysis has revealed that only one of thesemutants is tagged with a T-DNA insert.Copyright 1994, 1999 AcademicPress Arabidopsis thaliana, root, development, T-DNA insertion mutagenesis, endodermis, elongation 相似文献