A role for Fas in negative selection of thymocytes in vivo

To seek information on the role of Fas in negative selection, we examined subsets of thymocytes from normal neonatal mice versus Fas-deficient lpr/lpr mice injected with graded doses of antigen. In normal mice, injection of 1-100 microg of staphylococcal enterotoxin B (SEB) induced clonal elimination of SEB-reactive Vbeta8+ cells at the level of the semi-mature population of HSAhi CD4+ 8- cells found in the thymic medulla; deletion of CD4+ 8+ cells was minimal. SEB injection also caused marked elimination of Vbeta8+ HSAhi CD4+ 8- thymocytes in lpr/lpr mice. Paradoxically, however, elimination of these cells in lpr/lpr mice was induced by low-to-moderate doses of SEB (相似文献   

Requirement for p56lck tyrosine kinase activation in T cell receptor-mediated thymic selection

The nonreceptor protein tyrosine kinase p56lck (Lck) serves as a fundamental regulator of thymocyte development by delivering signals from the pre-T cell receptor (pre-TCR) that permit subsequent maturation. However, considerable evidence supports the view that Lck also participates in signal transduction from the mature TCR. We have tested this conjecture by expressing a dominant-negative form of Lck under the control of a promoter element (the distal lck promoter) that directs high expression in CD4+CD8+ thymocytes, mature thymocytes, and peripheral T cells, thereby avoiding, complications that result from the well-documented ability of dominant-negative Lck to block very early events in thymocyte maturation. Here we report that expression of the catalytically inactive Lck protein at twice normal concentrations inhibits thymocyte positive selection by as much as 80%, while leaving other aspects of cell maturation intact. This effect was studied in more detail in mice simultaneously bearing the male-specific H-Y alpha/beta TCR transgene and ovalbumin-specific DO10 alpha/beta TCR transgene, where even equimolar expression of the dominant-negative Lck protein substantially vitiated the positive selection process. Although deletion of H-Y alpha/beta thymocytes proceeded normally in male mice despite the presence of catalytically inactive Lck, modest inhibition of superantigen-mediated deletion was in some cases observed. These data further implicate Lck in the propagation of all TCR-derived signals, and indicate that even very modest deficiencies in the representation of functional Lck molecules could in humans, profoundly alter the character of the peripheral TCR repertoire.     

Homeostatic drive counteracting selection for positive and negative phototaxis and geotaxis in Drosophila pseudoobscura

Tested in Hirsch-Hadler mazes, Drosophila pseudoobscura is on the average geoneutral and photoneutral. Strongly geo- and photopositive and geo- and photonegative populations can be obtained by artificial selection, but upon relaxation of the selection they tend to relapse toward neutrality. This genetic homeostasis is due to natural selection favoring neutrality. Experiments are described in which artificial selection for positivity and for negativity was deliberately made so weak that it only counterbalanced the nautral selection or "homeostatic drive" and the effects of cross-migration. Under these conditions, the behavior of the population artificially selected for positivity diverges only slightly from that of the population selected for negativity, but, at least in females, both populations move closer to neutrality.     

Differences in the level of expression of class I major histocompatibility complex proteins on thymic epithelial and dendritic cells influence the decision of immature thymocytes between positive and negative selection

Both positive and negative selection of immature T cells rely on engagement of their antigen-specific receptors (TCR) by peptide in association with proteins encoded in the major histocompatibility complex (MHC) protein. The decision made between these two outcomes seems to be determined by the number of TCR engaged by peptide-MHC complexes. It has been unclear how such a mechanism can be reconciled with evidence that positive and negative selection occur in different thymic compartments and are mediated by different antigen-presenting cells (APCs). In this study we demonstrate that the level of class I MHC protein is 10-fold higher on thymic dendritic cells, which mediate the negative selection of immature T cells, than on thymic epithelial cells, which mediate for positive selection. We also demonstrate that as little as a 3-fold increase in the level of a particular cognate peptide-MHC ligand is sufficient to result in negative rather than positive selection. The results suggest that quantitative differences in the level of expression of class I MHC proteins on thymic epithelial and dendritic cells contribute to the opposing roles these cells play in forming the repertoire of mature class I MHC restricted (CD8+) T cells.     

Specific recognition of thymic self-peptides induces the positive selection of cytotoxic T lymphocytes

Congenital alveolar proteinosis is a recently described cause of lung dysfunction and respiratory distress in term neonates. In several cases a deficiency or insufficiency of surfactant apoprotein B (SP-B) has been caused by a frameshift mutation in the gene encoding SP-B. Five full-term children in three unrelated families from The Netherlands are reported. Immunohistochemistry demonstrated large amounts of surfactant proteins A and C (SP-A and SP-C) and precursors in alveolar cells and in intra-alveolar material. Results were positive for antibovine SP-B antibody but negative for antipig SP-B1 antibody, most probably reflecting differences in the antibody specificity. The findings suggest abnormal SP-B function. In two sibs, no pre-SP-C was demonstrated in the alveoli, although it was found in considerable amounts in alveolar cells. One such case has previously been reported. In two families, the parents were heterozygous for the 121 ins 2 mutation in the SP-B gene. Our findings suggest that congenital alveolar proteinosis may result from abnormalities in one or more of the surfactant proteins.     

Target selection difficulty, negative priming, and aging.

It has been recently suggested that the presence of identity negative priming effects in old adults could occur when there is substantial processing of the distracting information in a selective attention task (J. M. Kieley & A. A. Hartley, 1997). In three experiments, using a letter identification task, it was found that making target selection more difficult increased the magnitude of the negative priming effect to a similar extent in both young and old adults. Moreover, the size of the negative priming effect did not differ between young and elderly participants. These results are discussed with respect to the issue of age-related deficits in the mechanisms underlying negative priming. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

The thymic repertoire of neuroendocrine-related self antigens: biological role in T-cell selection and pharmacological implications

Thymic epithelium, including nurse cells (TEC/TNC), as well as other thymic stromal cells (macrophages and dentritic cells), express a repertoire of polypeptide belonging to various neuroendocrine protein families (such as the neurophypophysial, tachykinin, neurotensin and insulin families). A hierarchy of dominance exists in the organization of the thymic repertoire of neuroendocrine precursors. Oxytocin (OT) is more expressed in the TEC/TNC than vasopressin (VP); insulin-like growth factor 2 (IGF-2) thymic expression predominates over IGF-1, and much more over (pro)insulin. Thus, OT was proposed to be the self antigen of the neurohypophysial family, and IGF-2 the self antigen precursor of the insulin family. The dual role of the thymus in T-cell life and death is recapitulated at the level of the thymic neuroendocrine protein repertoire. Indeed, thymic polypeptides behave as accessory signals involved in T-cell development and positive selection according to the cryptocrine model of signaling. Moreover, thymic neuroendocrine polypeptides are the source of self antigens presented by thymic MHC molecules to developing pre-T cells. This presentation might induce the negative selection of T cells bearing a randomly rearranged antigen receptor (TCR) oriented against neuroendocrine families. Using an animal model of autoimmune type 1 diabetes (BB rat), we have shown a defect in intrathymic expression of the self antigen of the insulin family (IGF-2) and in IGF-2-mediated T-cell education to recognize and tolerate the insulin family. Altogether these studies have enlightened the crucial role played by the thymus in the induction of the central self tolerance of neuroendocrine families. The tolerogenic properties of thymic self peptides could be used in a novel type of vaccination for the prevention of autoimmune diseases.     

Coexisting undifferentiated thymic carcinoma and thymic carcinoid tumor

Thymic carcinoma is an unusual neoplasm, and the undifferentiated type is rare. Thymic carcinoid is also rare. This report describes a patient with coexisting undifferentiated thymic carcinoma and a carcinoid tumor. Both lesions were completely excised. The carcinoid cells showed argyrophilic granules by Grimelius' method and immunoreactivity against neuron-specific enolase, whereas the undifferentiated carcinoma cells were negative for argyrophilic stain and immunostaining against neuron-specific enolase.     

Direct visualization of thymocyte apoptosis in neglect, acute and steady-state negative selection

During thymocyte differentiation, the majority of the developing cells die in situ by apoptosis and are subsequently removed by macrophages. DNA fragmentation is one of the hallmarks of apoptosis and can be detected in situ by TdT-mediated dUTP-biotin nick end labeling (TUNEL). We used TUNEL combined with immunohistology to determine the sites of thymocyte apoptosis in mice transgenic for a TCR (F5) which recognizes a peptide (NP68) of the influenza virus nucleoprotein (NP) presented on the MHC class I H-2Db molecule. Apoptosis due to neglect was studied in F5 mice expressing a neutral MHC haplotype (F5/H-2q) and in beta 2-microglobulin-deficient F5 mice (F5/ beta 2m+). In both cases, the frequency of apoptotic cells was similar to that seen in F5/H-2b mice and non-transgenic C57BI/10 mice. Antigen-induced apoptosis was studied in F5 mice after i.p. Injection of the cognate NP68 peptide and in F5/NP double-transgenic mice. Three hours after peptide injection, apoptosis was high throughout the thymus cortex and clusters of apoptotic cells formed due to tissue macrophage uptake, whereas the thymic medulla remained unaffected. Massive recruitment of inflammatory cells into the thymus was seen as early as 1 h after peptide injection. Nine hours after peptide injection changes were apparent in the cortical epithelium and, by 4 days, the cortical network had collapsed to give scattered, compacted epithelial cells. In contrast, in F5/NP double-transgenic mice, thymocyte apoptosis induced by cognate self-peptide was localized at the cortico-medullary junction with little change seen in the epithelium of the cortex.     

Double-positive thymocytes resistant to antigen-MHC-induced negative selection lack active caspase

The aim of the current study was to elucidate the synergism of dietary calcium restriction and exhaustive exercise in the antioxidant enzyme system of rat soleus muscle, and to investigate the involvement of neutrophils in exercise-induced muscle damage. Forty-eight male Wistar rats were assigned to the following groups: control (C) or calcium-restricted [1 month (1 M) or 3 months (3 M)]. Each group was subdivided into acutely exercised or non-exercised groups. Soleus muscle from each rat was analysed to determine the levels of antioxidant enzymes [Mn-superoxide dismutase (SOD), Cu, Zn-SOD, glutathione peroxidase (GPX), and catalase (CAT)]. Dietary calcium restriction resulted in calcium deficiency and upregulated the antioxidant enzymes examined except GPX. Conversely, exhaustive exercise significantly decreased GPX and CAT, but not SODs activities in the calcium-restricted (1 M and/or 3 M) rats. Contents of immunoreactive Mn-SOD and Cu,Zn-SOD were only increased in the 3 M rats. During calcium restriction, the mRNA expression of both forms of SOD showed initial upregulation, followed by downregulation. Exhaustive exercise significantly increased the mRNA expressions only in the 3 M rats. Moreover, exhaustive exercise markedly increased myeloperoxidase activity in soleus muscles from the 1 M and 3 M rats compared with the C rats, and significantly enhanced the ability of neutrophils to generate superoxide in the 3 M rats. The results demonstrate that dietary calcium restriction upregulates certain antioxidant enzyme activities in rat soleus muscle, indicating an enhanced resistance to potential increases in intracellular reactive oxygen species. The results also suggest that exhaustive exercise may cause oxidative damage in soleus muscle of calcium-deficient rats through the activation of neutrophils.     

Influence of probe-trial selection on the location negative priming effect.

Using the location variant of the typical negative priming procedure, participants were cued (100% reliable) before (Experiment 1) or after (Experiment 2) the prime trial as to whether a distractor would or would not accompany the target on the probe trial. The crucial results were that on cued trials, the predictable absence of the probe-trial distractor, but not its cued presence, produced the removal of the negative priming effect (disengagement), and that this disengagement of the priming process, motivated by the predictable absence of a probe-trial distractor, could take place on-line. These findings demonstrated the "selection-state" dependency (probe trial) of the location negative priming process, supporting inhibition-based and episodic retrieval models in their contention that the ultimate function of this process is to enhance the efficiency of future distractor processing, and hence selection. The disengagement results revealed an adaptive feature of a process that can be detrimental or irrelevant to upcoming processing. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

The role of gender and negative affectivity in stressor appraisal and coping selection.

Individual differences in stress may arise from many sources. This study investigated the role of gender and negative affectivity (NA) in stressor appraisal and coping selection. Differential exposure to stressors was controlled by requiring participants to rate the stressfulness of identical hypothetical scenarios. As predicted, women rated the scenarios as more stressful than men, and perceptions of stressfulness increased with participant NA. Women endorsed the use of emotion-focused coping strategies more than men, even when perceived stressfulness was controlled. NA predicted use of both emotion- and avoidance-focused coping, although only the latter association remained significant after controlling for stressor appraisals. Gender × NA interaction effects were not significant. Implications for the prediction and management of stress are discussed. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Solidification of undercooled Fe-Cr-Ni alloys: Part III. Phase selection in chill casting

In Parts I and II of this series of articles, it was shown that a range of levitation-melted Fe-Cr-Ni alloys, both hypoeutectic and hypereutectic, all solidified with the hypereutectic phase (bcc) as their primary phase, except for the hypoeutectic alloys at low undercoolings. In this article, the effect of heat extraction on phase formation is studied by chill casting the undercooled alloys before nucleation. Two of the previously studied alloys are examined; one hypoeutectic and the other hypereutectic. Chill substrates employed were copper, stainless steel, alumina, zirconia, and a liquid gallium-indium bath. Contrary to the case of levitation melting and solidification, it is found that the dominant primary phase to solidify in both alloys, independent of chill substrate, is the hypoeutectic phase (fcc). It is concluded that chilling the undercooled melt results in nearly concurrent nucleation of bcc and fcc. Two different mechanisms are considered as possible explanations of the subsequent fcc phase selection during growth. These are termed “growth velocity” and “phase stability” mechanisms. Evidence favors a phase stability mechanism, in which the bcc phase massively transforms to fcc early in solidification so that fcc then grows without competition. It is suggested that this mechanism may also explain structures observed in welds and other rapid solidification processes.     

Crucial role of Jak3 in negative selection of self-reactive T cells

The stimulation of cardiomyocyte guanylyl cyclase by nitric oxide (NO)-donor drugs was examined before and after exposure of these cells to the NO-donor drugs: S-nitroso-d,l-acetylpenicillamine (SNAP) and sodium nitroprusside (SNP). Short- (2-hr) and long-term (24-hr) exposure attenuated the maximal stimulation of GC by either SNAP or SNP by up to 80% ("desensitization"). However this "desensitization" of the myocardial GC was atypical in nature in that the reduction in maximal NO-stimulated GC activity was associated with an increase in the affinity of the GC towards either NO-donor, a finding not as yet reported. There was also evidence of "cross-desensitization" of GC (e.g., SNAP exposure decreasing the stimulatory effect of SNP). Further, this is the first time that SNAP-induced desensitization of GC has been observed.     

Defective T cell receptor signaling and CD8+ thymic selection in humans lacking zap-70 kinase

We have previously described a type of selective T cell deficiency (STD) characterized by persistent infections reminiscent of severe combined immunodeficiency. We show here that STD patients carry a mutation of zap-70, resulting in loss of the activity of this kinase. The thymi of zap-70-/- patients show the presence of CD4+CD8+ cells in the cortex; however, only CD4, not CD8, single-positive cells are present in the medulla. Peripheral CD4+ T cells from the zap-70-/- patients exhibit markedly reduced tyrosine phosphorylation, fail to produce interleukin-2, and do not proliferate in response to T cell receptor stimulation by mitogens or antigens. Thus, Zap-70 kinase appears to be indispensable for the development of CD8 single-positive T cells as well as for signal transduction and function of single-positive CD4 T cells.