bluuebunny/crossref_metadata_2025 huggingface dataset:
Variabilité des pratiques funéraires et différenciation sociale dans le Néolithique ancien danubien
Authors: Jeunesse Christian
Published: March 2010

In the early seventies, results from the Nitra cemetery reinforced the idea that there was little social differentiation in the Bandkeramik. The corpus of published Danubian Early Neolithic funerary data has considerably increased since then. Important series are now available for the whole Bandkeramik sequence and most of the distribution area. Their study shows that the Nitra model is only valid within a limited geographical and chronological framework. The major cemeteries discovered since 1960 from Bavaria to the Meuse reveal a gradual development towards a more complex society, with a higher degree of both horizontal and vertical differentiation. The latest cemeteries are characterized by a small group of very rich graves which stand out clearly from the remainder. Within this group, the recurrent presence of child burials with the same categories of grave-good as the adults strongly suggests hereditary transmission of status. In the second half of the Early Neolithic, this model coexists with the Nitra model which still determines how the large majority of the population was treated. This development towards a marked increase in inequality is corroborated by changes in settlement. As is attested by the considerable evidence for violent death recovered over recent years, the resulting more complex society appears torn by internal contradictions. This takes us far from the bucolic image that still implicitly underlies most overviews of Danubian Early Neolithic society.
Foyer domestique ou foyer annexe. Les modes d'occupation de l'espace des Magdaléniens d'Étiolles
Authors: Olive Monique
Published: March 2010

Les fouilles extensives conduites sur certains gisements magdaléniens du Bassin parisien mettent au jour progressivement de véritables campements. Le schéma général qui se dégage est celui d'une juxtaposition d'unités d'occupation composées d'un foyer associé à une densité variable de vestiges. Parmi les questions que pose l'interprétation de ces campements figure la reconnaissance des habitations en l'absence de tout élément de construction. A Etiolles, la démonstration s'appuie sur la comparaison de deux unités d'occupation particulières choisies pour leur absence d'organisation évidente. La démarche, de nature inductive, prend en compte l'ensemble des vestiges conservés dont l'analyse spatiale met en évidence des comportements spécifiques dans le mode d'occupation de l'espace et dans l'organisation des activités. Ces traits de comportements sont mis en relation avec la fonction de ces unités dans le campement.
Le site néolithique moyen de Derrière-les-Prés à Ernes (Calvados)
Authors: San Juan Guy , Dron Jean-Luc
Published: March 2010

The Derrière-les-Prés passage grave at Ernes (Calvados) is situated 25 km south of Caen, on the western slope of the Laizon

valley. It was partially explored around 1840 par Mr Bellivet, member of the Société des Antiquaires de Normandie. The recent large

scale excavation ended in 1990. Under the remains of the levelled limestone dry stone walled monument and its rubble, artefacts and

features corresponding to two anterior occupation phases were fossilized in the ancient land surface. The oldest occupation can be clearly attributed to the Cerny civilisation by a concentration of pottery and lithic artefacts. The radiocarbon dating of a surface hearth (4740- 4070 B.C.) preserved under the monument enables us, with some reservations, to associate the feature to the Cerny period. The second occupation before the use of the grave is attributed to the Middle Neolithic II, by scarcer lithic and pottery artefacts.

The radiocarbon dates from two hearths in shallow pits under the rubble (4072-3797 and 4080-3743 B.C.) enable us to attribute them

either to a domestic occupation prior to the grave, or to an occupation associated to its erection and use.

The monumental phase begins with the laying out of the funerary chamber, probably by temporary pegging, marked in the ancient ground level by a circle of post hole stones. This state has not been dated, neither has the construction of a double faced wall. The later consolidation however, consisting of an annular mass of stones, is dated by animal bones abandoned in the stone work (3950-3528 B.C.). The grave use is certified by a bone date (3950-3190 B. C.) from a single skeleton, that of a dwarf, found partially in situ. A pedestal based dish found in the sepulchral area confirms that the passage grave was in use during the Middle Neolithic II. The mid-neolithic environment is mainly defined by anthracological analysis. The results show selective use of large pines and oaks. The presence of ash shows use of valley bottom resources. Hazel and Pomaceaous trees mark the transition to the agrarian space in which we find cereal growing (wheat), and classical animal husbandry (Bovidae, Ovicapridae and pigs) in a not well defined area.
Circulation des silex dans le nord de l'Aquitaine au Paléolithique supérieur : l'occupation de l'espace par les derniers chasseurs-cueilleurs
Authors: Demars Pierre-Yves
Published: March 2010

Pendant le Paléolithique supérieur, dans le nord de l'Aquitaine, les silex ont été transportés sur de grandes distances. Ces circulations de matières premières mettent en relation des régions allant de la Gironde au Massif central suivant un axe est-ouest, le long du bassin de la Dordogne. Ce fait complète le modèle d'occupation de l'espace proposé par R. K White : les sites de la basse vallée de la Vézère sont liés à des abattages massifs de rennes lors de leurs migrations et sont des habitats de mauvaise saison. Dans les régions voisines, les autres concentrations de sites semblent plutôt être en majorité des habitats de belle saison. Ce modèle disparaît au Mésolithique avec l'amélioration climatique holocène.
Essai d'interprétation du site paléolithique inférieur de Soucy 1 (Yonne)
Authors: Lhomme Vincent , Connet Nelly , Bémilli Céline , Chaussé Christine , Beyries Sylvie , Guérin Claude
Published: March 2010

Soucy 1 is the first of six Middle Pleistocene sites found in fluvial context on an ancient terrace of the Yonne river at Soucy (North Burgundy), in a stratigraphic sequence cautiously attributed to the Holsteinian interglacial. It was excavated during a salvage operation in 1995 and has yielded more than 2 500 remains (lithic, fauna and pebbles). Spatial analysis reveals areas of flaking activities, concentrations of faunal remains and one concentration of unretouched pebbles, all indicating differential activities. These elements allow us to propose an interpretation about the site function.
Figures inédites de la grotte de Rouffignac
Authors: Plassard Frédéric , Plassard Jean
Published: March 2010

L'observation répétée des parois et plafonds de la cavité a depuis quelques années donné lieu à la découverte de nouvelles œuvres justifiant des relevés et leur publication. Bien que souvent incomplètes, ces quelques figures éclairent d'un jour nouveau la perception de l'organisation du décor pariétal de ce site. Elles démontrent qu 'en dépit de son apparente clarté résultant de l'ampleur du réseau, ce dispositif pariétal est une bonne illustration de l'existence de particularismes locaux ou chronologiques.
Premières données sur le traitement des corps humains à la transition du Néolithique récent et du Néolithique final dans le Bas-Rhin : dimensions culturelles
Authors: Blaizot Frédérique , Boës Xavier , Lalaï Dominique , Le Meur Nelly , Maigrot Yolaine
Published: March 2010

Human remains have been found in six round pits on the site of Reichstett-Mundolsheim ; five of these pits contain inhumations, one of which is dual, dating from the end of the Late Neolithic II. In the last one, eleven bodies burnt altogether, belong to the begining of the Sub Neolithic period. In this paper, the detailed study of the different rituals which have been noticed (burial practices and cremation process) was carried out with archaeo-anthropological methods. Cremation is the earliest evidence for the treatment of the corpses concerning the Sub Neolithic in Alsace, and the practice recognized is radically different from what we meet in the west and south parts of this area. But in other hand, parallels can be found in Eastern Europe.
Les productions lamellaires dans l'Aurignacien de l'abri Pataud, Les Eyzies-de-Tayac (Dordogne)
Authors: Chiotti Laurent
Published: March 2010

Durant l'Aurignacien, diverses techniques ont été développées afin de produire des lamelles. Dans la séquence aurignacienne de l'abri Pataud (9 niveaux), ces différentes chaînes opératoires ont pu être mises en évidence et leur importance précisée pour chaque stade de l'Aurignacien. La production de lamelles à partir des grattoirs aurignaciens (carénés et à museau) est présente durant toute la séquence. Comme c'est généralement le cas, la production à partir des burins busqués se situe quasi exclusivement dans l'Aurignacien évolué, cependant, une chaîne opératoire identique a pu être décrite dans l'un des niveaux de l'Aurignacien ancien. Dans l'Aurignacien ancien, les petites lamelles qui sont produites à partir des grattoirs sont plutôt des lamelles rectilignes ou courbes, alors que dans l'Aurignacien évolué elles sont essentiellement courbes et torses, qu 'elles soient produites à partir des grattoirs ou des burins. Une production de lamelles de plus grandes dimensions à partir de nucléus prismatiques existe durant toute la séquence, même si elle est toujours faible. L'outillage lamellaire est peu développé, à l'exception des lamelles Dufour du niveau 8 (Aurignacien évolué), et ce malgré la grande quantité de lamelles produite, ce qui pose la question de leur utilisation brute.
Les céramiques communes des Campaniformes européens
Authors: Besse Marie
Published: March 2010

Die Glockenbecherzeit ist als Einheit immer noch ziemlich schlecht definiert, obwohl es schon zahlreiche Arbeiten über diese Epoche gibt. Ihr Ausbreitungsareal ist so gross, dass man sich einige Fragen stellen muss : Haben wir es mit einer Ideologie zu tun ? Mit Völkern ? Mit einer Völkerwanderung ? Betraf diese mehrere Menschengruppen ? Gab es nur eine Wanderungswelle oder mehrere ? In welche Richtungen ? Wir werden versuchen, diese Fragen anhand der glockenbecherzeitlichen Gebrauchskeramik zu beantworten. Diese Keramik wurde beschrieben und in Typen eingeteilt, es wurden im Ganzen 83 verschiedene Typen ausgemacht. 800 Fundstellen wurden untersucht, sie verteilen sich auf 11 Länder : Belgien, Deutschland, Frankreich, Holland, Italien, Österreich, Polen, Schweiz, Slowakien, Tschechische Republik und Ungarn. Die Keramikgruppen weisen in den verschiedenen Gegenden so grosse Unterschiede auf, dass man drei grosse Kulturgebiete erkennen kann : das östliche Gebiet, dans nördliche Gebiet und das südliche Gebiet.
Centres de production et diffusion des silex bédouliens au Chasséen
Authors: Léa Vanessa
Published: March 2010

At the 5th and 4th millenium BC, in a Chassean context, the distribution of Bedoulian flint of Haute-Provence (Lower Cretaceous), extents impressively and is considered as an absolutely original phenomenon. The lithic assemblages are mainly composed of products made from this raw material being exported. But if numerous studies dealing with consumer sites have been undergone these last years, the production environment is still totally misunderstood. Recent discoveries in the production area give a new impetus to research on this essential subject for a better understanding of Chassean communities. The approach of two new production sites, Les Trois Termes (Gordes) and Rocalibert (Piolenc) gives the opportunity to study the increasing specializations and the context of production during the Chassean.
Propriétés combustibles des ossements : données expérimentales et réflexions archéologiques sur leur emploi dans les sites paléolithiques
Authors: Théry-Parisot Isabelle , Costamagno Sandrine
Published: March 2010

The hypothesis of bone being used as fuel is put forward to explain the abundance of burnt bones inside bone assemblages. This utilization is often compared with the lack of wood in the surroundings of sites. Yet, utilization of bone seems to be attested all along the Upper Palaeolithic aside ambient climatic conditions. As a matter of fact, some of the occupations are contemporaneous with temparate phases when palaeoenvironmental data show the existence of a forest environment. Therefore, it is possible to consider a well-reasoned utilization of this fuel, regardless the available ligneous biomass but deeply connected to local household activities. Only the knowledge of the combustible properties of bone may bring part of an answer about this preferential utilization as a hypothesis, according to the function of the hearth. A vast experimental programme is undertaken for this purpose. The experiments on more than 120 kg of bone were carried out along with intrinsic and extrinsic parameters which govern combustion. The results allow us to determine precisely the bone combustible properties along with their histological nature, the level of humidity and the fractures. We can also distinguish bone assemblages strictly being used as fuel from those having fortuitous combustion marks. Therefore, the properties of bone and wood can be paralleled and so it is possible to define more clearly the purpose of a preferential utilization of bone and to propose a model of fuel management appropriate to energy needs of each group.
Architectural Approaches for HL7-based Health Information Systems Implementation
Authors: López D. M. , Blobel B.
Published: March 2010

Objective: Information systems integration is hard, especially when semantic and business process interoperability requirements need to be met. To succeed, a unified methodology, approaching different aspects of systems architecture such as business, information, computational, engineering and technology viewpoints, has to be considered. The paper contributes with an analysis and demonstration on how the HL7 standard set can support health information systems integration.

Methods: Based on the Health Information Systems Development Framework (HIS-DF), common architectural models for HIS integration are analyzed. The framework is a standard-based, consistent, comprehensive, customizable, scalable methodology that supports the design of semantically interoperable health information systems and components.

Results: Three main architectural models for system integration are analyzed: the point to point interface, the messages server and the mediator models. Point to point interface and messages server models are completely supported by traditional HL7 version 2 and version 3 messaging. The HL7 v3 standard specification, combined with service-oriented, model-driven approaches provided by HIS-DF, makes the mediator model possible. The different integration scenarios are illustrated by describing a proof-of-concept implementation of an integrated public health surveil-lance system based on Enterprise Java Beans technology.

Conclusion: Selecting the appropriate integration architecture is a fundamental issue of any software development project. HIS-DF provides a unique methodological approach guiding the development of healthcare integration projects. The mediator model – offered by the HIS-DF and supported in HL7 v3 artifacts – is the more promising one promoting the development of open, reusable, flexible, semantically interoperable, platform-independent, service-oriented and standard-based health information systems.
Bioorthogonal metabolic glycoengineering of human larynx carcinoma (HEp-2) cells targeting sialic acid
Authors: Homann Arne , Qamar Riaz-ul , Serim Sevnur , Dersch Petra , Seibel Jürgen
Published: March 2010

Sialic acids are located at the termini of mammalian cell-surface glycostructures, which participate in essential interaction processes including adhesion of pathogens prior to infection and immunogenicity. Here we present the synthesis and bioorthogonal metabolic incorporation of the sialic acid analogue N-(1-oxohex-5-ynyl)neuraminic acid (Neu5Hex) into the cell-surface glycocalyx of a human larynx carcinoma cell line (HEp-2) and its fluorescence labelling by click chemistry.
Innovative Strategies for Nursing Education Program Evaluation
Authors: Story Lachel , Butts Janie B. , Bishop Sandra B. , Green Lisa , Johnson Kathy , Mattison Haley
Published: March 2010

Nursing programs are mandated by accreditation bodies to report data significant to program quality and outcomes. The history at one school of nursing in the southern United States revealed the program evaluation committee experienced roadblocks in retrieving such information. Creative approaches were adopted to overcome some of the barriers to program evaluation, including the use of more technological-based approaches to engage alumni who embrace this technology as a way of life. Among the many advantages of these approaches were convenience, ease of administration and analysis, cost effectiveness, and more meaningful data. The advantages far outweighed the few disadvantages incurred, with the most prominent being potential sampling bias.
Collaborative Action Research: Implementation of Cooperative Learning
Authors: Smith-Stoner Marilyn , Molle Mary E.
Published: March 2010

Nurse educators must continually improve their teaching skills through innovation. However, research about the process used by faculty members to transform their teaching methods is limited. This collaborative study uses classroom action research to describe, analyze, and address problems encountered in implementing cooperative learning in two undergraduate nursing courses. After four rounds of action and reflection, the following themes emerged: students did not understand the need for structured cooperative learning; classroom structure and seating arrangement influenced the effectiveness of activities; highly structured activities engaged the students; and short, targeted activities that involved novel content were most effective. These findings indicate that designing specific activities to prepare students for class is critical to cooperative learning.
Coats' Disease and Neovascular Glaucoma in a Child With Neurofibromatosis
Authors: Chiu Shin-Lin , Chen San-Ni , Chen Yan-Ting , Chen Pao-Ju
Published: March 2010

This report describes an extremely rare case of Coats' disease and neovascular glaucoma in a child with neurofibromatosis 1. Intraocular pressure control and vision restoration were achieved with intravitreal injection with bevacizumab, hemiretinal laser photocoagulation, peripheral retinal cryotherapy, and trabeculectomy with mitomycin C.
Malignant Mixed Tumor of the Lacrimal Gland in a Teenager
Authors: Giliberti Francesca M. , Shinder Roman , Bell Diana , Savar Aaron M. , Sacks Justin M. , Levine Nicholas B. , Esmaeli Bita
Published: March 2010

A 17-year-old boy presented with a painless, right superotemporal orbital mass. Imaging demonstrated a lacrimal gland mass extending into the temporalis muscle through the lateral orbital wall. The patient underwent an orbital exenteration. Histopathology revealed malignant mixed tumor (carcinoma ex-pleomorphic adenoma) of the lacrimal gland with perineural and vascular invasion.
Silicone-Eyelid Closure to Improve Vision in Deeply Amblyopic Eyes
Authors: Hakim Ossama M. , El-Hag Yasser Gaber , Samir Ahmed
Published: March 2010

Background:

Deep amblyopia is a recognized cause of monocular visual deficit in children with a high rate of poor compliance with conventional occlusion therapy. This study evaluates a new occlusion technique that can be used for children with failed amblyopia treatment.

Methods:

In a prospective study, surgical silicone-eyelid closure was done for 15 children (age: 4 to 6 years; median age: 5.2 years) with a history of deep amblyopia and poor compliance with occlusion methods. In this technique, the good eye was closed by passing one limb of a double armed 5-0 polypropylene suture from one eyelid margin through a silicone sleeve and through the corresponding eyelid margin. The other polypropylene limb was passed behind the silicone sleeve and then through the opposite eyelid margin to form a barrier between the sleeve and the cornea. After 2 to 4 weeks, the suture was cut and the silicone sleeve was removed.

Results:

Preoperative visual acuity ranged between counting fingers at 1 meter and counting fingers at 4 meters. Following eyelid closure, visual acuity was improved to between 20/40 and 20/200 for 12 patients, whereas three patients did not improve. Complications were seen in 8 patients: marginal eyelid irritation and erythema in 6 and suture break in 2.

Conclusion:

Silicone-eyelid closure can be used safely and effectively for management of deep amblyopia. This technique affords a good option to restore vision in a deeply amblyopic eye and grants the child a lifetime of useful vision.

[J Pediatr Ophthalmol Strabismus 2010;47:157–162.]
BTN1A1, the Mammary Gland Butyrophilin, and BTN2A2 Are Both Inhibitors of T Cell Activation
Authors: Smith Isobel A. , Knezevic Brittany R. , Ammann Johannes U. , Rhodes David A. , Aw Danielle , Palmer Donald B. , Mather Ian H. , Trowsdale John
Published: March 2010

Butyrophilin (BTN) genes encode a set of related proteins. Studies in mice have shown that one of these, BTN1A1, is required for milk lipid secretion in lactation, whereas butyrophilin-like 2 is a coinhibitor of T cell activation. To understand these disparate roles of BTNs, we first compared the expression and functions of mouse Btn1a1 and Btn2a2. Btn1a1 transcripts were not restricted to lactating mammary tissue but were also found in virgin mammary tissue and, interestingly, spleen and thymus. In confirmation of this, BTN1A1 protein was detected in thymic epithelial cells. By contrast, Btn2a2 transcripts and protein were broadly expressed. Cell surface BTN2A2 protein, such as the B7 family molecule programmed death ligand 1, was upregulated upon activation of T cells. We next examined the potential of both BTN1A1 and BTN2A2 to interact with T cells. Recombinant Fc fusion proteins of murine BTN2A2 and, surprisingly BTN1A1, bound to activated T cells, suggesting the presence of one or more receptors on these cells. Immobilized BTN-Fc fusion proteins, but not MOG-Fc protein, inhibited the proliferation of CD4 and CD8 T cells activated by anti-CD3. BTN1A1 and BTN2A2 also inhibited T cell metabolism, IL-2, and IFN-γ secretion. Inhibition of proliferation was not abrogated by exogenous IL-2 but could be overcome following costimulation with high levels of anti-CD28 Ab. These data are consistent with a coinhibitory role for mouse BTNs, including BTN1A1, the BTN expressed in the lactating mammary gland and on milk lipid droplets.
Hemokinin-1 Activates the MAPK Pathway and Enhances B Cell Proliferation and Antibody Production
Authors: Wang Wei , Li Qian , Zhang Jun , Wu Hounan , Yin Yanhui , Ge Qing , Zhang Yu
Published: March 2010

Hemokinin 1 (HK-1) is a substance P-like tachykinin peptide predominantly expressed in non-neuronal tissues. In addition to a prominent function in lymphoid development, recent studies indicate a potential role for HK-1 in immunoregulation. The current study was focused on its action on mature B cells. Despite the negligible effect on its own, HK-1 exhibited a profound influence on B cell activation elicited by several classical signals, including LPS stimulation, BCR cross-linking, and CD40 ligation. Cells therefore showed enhanced proliferation, survival, and CD80/86 expression, and produced more IgM with a higher frequency of Ab-forming cells. Biochemical analysis revealed that HK-1 alone was sufficient to induce the activation of MAPKs and the expression of Blimp-1 and Xbp-1 in B cells. Nevertheless, costimulation with a known B cell activator resulted in much enhanced phosphorylation of MAPKs and transcriptional activation of Blimp-1 and Xbp-1. Overall, these data support that HK-1 provides an important costimulatory signal for B cell activation, possibly through synergistic activation of the MAPK pathway and induction of transcription factors critical for plasmacytic differentiation.
5-Lipoxygenase Activating Protein Signals Adipose Tissue Inflammation and Lipid Dysfunction in Experimental Obesity
Authors: Horrillo Raquel , González-Périz Ana , Martínez-Clemente Marcos , López-Parra Marta , Ferré Natàlia , Titos Esther , Morán-Salvador Eva , Deulofeu Ramon , Arroyo Vicente , Clària Joan
Published: March 2010

The presence of the so-called low-grade inflammatory state is recognized as a critical event in adipose tissue dysfunction, leading to altered secretion of adipokines and free fatty acids (FFAs), insulin resistance, and development of hepatic complications associated with obesity. This study was designed to investigate the potential contribution of the proinflammatory 5-lipoxygenase (5-LO) pathway to adipose tissue inflammation and lipid dysfunction in experimental obesity. Constitutive expression of key components of the 5-LO pathway, as well as leukotriene (LT) receptors, was detected in adipose tissue as well as in adipocyte and stromal vascular fractions. Adipose tissue from obese mice, compared with that from lean mice, exhibited increased 5-LO activating protein (FLAP) expression and LTB4 levels. Incubation of adipose tissue with 5-LO products resulted in NF-κB activation and augmented secretion of proinflammatory adipokines such as MCP-1, IL-6, and TNF-α. In addition, LTB4, but not LTD4, reduced FFA uptake in primary adipocytes, whereas 5-LO inhibition suppressed isoproterenol-induced adipose tissue lipolysis. In mice with dietary obesity, elevated FLAP expression in adipose tissue was paralleled with macrophage infiltration, increased circulating FFA levels, and hepatic steatosis, phenomena that were reversed by FLAP inhibition with Bay-X-1005. Interestingly, FLAP inhibition induced AMP-activated protein kinase phosphorylation in parallel with decreases in hormone-sensitive lipase activity and the expression and secretion of TNF-α and IL-6. Similar effects were observed in differentiated 3T3-L1 adipocytes incubated with either Bay-X-1005 or the selective LTB4 receptor antagonist U-75302. Taken together, these findings indicate that the 5-LO pathway signals the adipose tissue low-grade inflammatory state and steatogenic potential in experimental obesity.
Increased Foxp3+ Regulatory T Cells in Poly(ADP-Ribose) Polymerase-1 Deficiency
Authors: Nasta Francesca , Laudisi Federica , Sambucci Manolo , Rosado Maria M. , Pioli Claudio
Published: March 2010

Growing evidence is unveiling a role for poly(ADP-ribose) polymerase (PARP)-1 in the regulation of inflammatory/immune responses. In the current study, we investigated the effects of PARP-1 deficiency on regulatory T cell differentiation. Increased numbers of regulatory CD4+CD25+/Foxp3+ T cells were found in thymus, spleen, and lymph nodes of PARP-1 knockout (KO) mice compared with wild-type (WT) controls. The increased frequency of regulatory T cells in the periphery resulted in impaired CD4 cell proliferation and IL-2 production, which could be restored by CD25+ cell depletion. Phenotype and inhibitory functions of PARP-1 KO regulatory T cells were similar to WT cells, indicating that PARP-1 affects regulatory T cell differentiation rather than function. Purified naive CD4 cells from PARP-1 KO mice stimulated in vitro expressed forkhead box p3 mRNA at higher levels and generated a greater number of Foxp3+ cells (inducible regulatory T [iTreg] cells) than the WT counterpart. This finding was due to a higher rate of naive CD4 cell to Foxp3+ iTreg cell conversion rather than to higher resistance to apoptosis induction. Interestingly, PARP-1 deficiency did not affect retinoid-related orphan receptor-γt mRNA expression and differentiation of purified naive CD4 cells to Th17 cells. PARP-1 KO iTreg cells showed features similar to WT regulatory T cells, suggesting that modulation of PARP-1 during the immune response might be used to induce greater numbers of functional regulatory T cells. In conclusion, our findings represent the first evidence that PARP-1 can affect regulatory T cell differentiation and open new perspectives on potential targets for modulating immune responses.
Natural Exposure to Cutaneous Anthrax Gives Long-Lasting T Cell Immunity Encompassing Infection-Specific Epitopes
Authors: Ingram Rebecca J. , Metan Gökhan , Maillere Bernard , Doganay Mehmet , Ozkul Yusuf , Kim Louise U. , Baillie Les , Dyson Hugh , Williamson E. Diane , Chu Karen K. , Ascough Stephanie , Moore Steven , Huwar Theresa B. , Robinson John H. , Sriskandan Shiranee , Altmann Daniel M.
Published: March 2010

There has been a long history of defining T cell epitopes to track viral immunity and to design rational vaccines, yet few data of this type exist for bacterial infections. Bacillus anthracis, the causative agent of anthrax, is both an endemic pathogen in many regions and a potential biological warfare threat. T cell immunity in naturally infected anthrax patients has not previously been characterized, which is surprising given concern about the ability of anthrax toxins to subvert or ablate adaptive immunity. We investigated CD4 T cell responses in patients from the Kayseri region of Turkey who were previously infected with cutaneous anthrax. Responses to B. anthracis protective Ag and lethal factor (LF) were investigated at the protein, domain, and epitope level. Several years after antibiotic-treated anthrax infection, strong T cell memory was detectable, with no evidence of the expected impairment in specific immunity. Although serological responses to existing anthrax vaccines focus primarily on protective Ag, the major target of T cell immunity in infected individuals and anthrax-vaccinated donors was LF, notably domain IV. Some of these anthrax epitopes showed broad binding to several HLA class alleles, but others were more constrained in their HLA binding patterns. Of specific CD4 T cell epitopes targeted within LF domain IV, one is preferentially seen in the context of bacterial infection, as opposed to vaccination, suggesting that studies of this type will be important in understanding how the human immune system confronts serious bacterial infection.
The IL-7 Signaling Pathway Regulates Lymph Node Development Independent of Peripheral Lymphocytes
Authors: Chappaz Stéphane , Finke Daniela
Published: March 2010

Lymph node (LN) organogenesis is initiated by the interaction between hematopoietic lymphoid tissue inducer (LTi) cells and the mesenchymal organizer cells. Mice in which the IL-7 signaling pathway has been disrupted have a severe defect in LN development; however, the reasons underlying this defect are as yet unknown. In this study, we show that the overexpression of thymic stromal lymphopoietin (TSLP) increased LTi cell numbers and restored LN development in IL-7−/− and RAG2−/− γc−/− mice. The TSLP-mediated LN restoration was strictly dependent on LTi cells and independent of lymphocyte colonization. Increased LTi cell numbers in the LN anlagen of RAG2−/− γc−/− TSLP transgenic mice were associated with the restoration of organizer cells, suggesting that LTi cell number is a critical parameter for LN organogenesis. Our results shed light on the minimal cellular requirement for LN development during ontogeny. We show that the presence of LTi and organizer cells, but not of peripheral lymphocytes, is critical for LN development and persistence and further suggest that the IL-7 signaling pathway regulates LN organogenesis by controlling the size of the LTi cell pool.
ATP Activates Regulatory T Cells In Vivo during Contact Hypersensitivity Reactions
Authors: Ring Sabine , Enk Alexander H. , Mahnke Karsten
Published: March 2010

CD4+CD25+Foxp3+ regulatory T cells (Tregs) require activation to develop their full suppressive capacity. Similar to conventional T cells, Tregs can be activated via their TCRs; however, other means may be in place. We injected naive and nonactivated Tregs, being CD69−CD44lowCD62L+ into mice, and analyzed their phenotype after sensitization or challenge with the contact sensitizer 2,4,6-trinitro-1-chlorobenzene. We found that Tregs acquired an activated phenotype (CD69+CD44highCD62L−) in the draining lymph node after sensitization. In contrast, Ag challenge activated Tregs in the blood. This tissue-specific activation was induced by ATP, which was released at the respective tissue sites after sensitization or challenge, respectively. To demonstrate that activation was also essential for the induction of the suppressive function of Tregs, Tregs were treated with ATP receptor antagonists. In this study, we show that ATP receptor antagonists abrogated the suppressive effects of injected naive Tregs in contact hypersensitivity reactions. Thus, these data indicate that activation of Tregs via ATP in vivo provides a novel pathway of stimulating the suppressive function of Tregs.
SP1-Dependent Induction of CD39 Facilitates Hepatic Ischemic Preconditioning
Authors: Hart Melanie L. , Gorzolla Iris C. , Schittenhelm Jens , Robson Simon C. , Eltzschig Holger K.
Published: March 2010

Ischemia/reperfusion injury (IRI) of the liver is an important cause of hepatic dysfunction. Ischemic preconditioning (IP) is associated with adenosine-mediated tissue protection from subsequent IRI. Extracellular nucleotides (e.g., ATP) represent the main source for extracellular adenosine. Therefore, we hypothesized that phosphohydrolysis of ATP/ADP via the ectonucleoside triphosphate diphosphohydrolase-1 (CD39), conversion of ATP/ADP to AMP, mediates IP-dependent liver protection. We found that hepatic IP was associated with significant induction of CD39 transcript, heightened protein expression, and improved outcomes after IRI. Targeted gene deletion or pharmacological inhibition of CD39 abolished hepatoprotection by IP as measured by serum markers of liver injury or histology. Therapeutic studies to mimic IP with i.p. apyrase (a soluble ectonucleoside triphosphate diphosphohydrolase, NTPDase) in the absence of IP attenuated hepatic injury after IRI. In additional in vivo studies, small interfering RNA treatment was used to achieve repression of the transcription factor Sp1, known to be implicated in CD39 transcriptional regulation. In fact, Sp1 small interfering RNA treatment was associated with attenuated CD39 induction and increased hepatic injury in vivo. Our data suggest a Sp1-dependent regulatory pathway for CD39 during hepatic IP. These studies reveal a novel role of CD39 in hepatic protection and suggest soluble apyrase for the treatment of liver ischemia.
T Cell-Independent IgA Class Switch Recombination Is Restricted to the GALT and Occurs Prior to Manifest Germinal Center Formation
Authors: Bergqvist Peter , Stensson Anneli , Lycke Nils Y. , Bemark Mats
Published: March 2010

Recently, we reported that CD40−/− mice, exhibiting exclusively T cell-independent IgA class switch recombination (CSR), demonstrated near normal levels of IgA plasma cells in the gut lamina propria (LP), despite the complete lack of germinal centers (GCs). In this study, we have extended our analysis focusing on how to reconcile these findings using flow cytometry and molecular markers for IgA CSR. In agreement with our previous results with small intestinal LP, the colon LP was found to host IgA CSR only when lymphoid follicles were present. Thus, no IgA CSR was observed in the nonorganized colon LP. By contrast, the Peyer’s patch (PP) was the dominant IgA CSR site in both CD40−/− and wild type (WT) mice, and they both hosted similar levels of mRNA expression for B cell activating factor of the TNF family, a proliferation inducing ligand, and inducible NO synthase, potential switch-factors for IgA. Unexpectedly, we found that PP B cells undergoing IgA CSR were GL7-intermediate. These cells had not undergone somatic hypermutations (SHMs), whereas GL7-high cells in WT PP, which exhibited GCs, were heavily mutated. Moreover, IgA plasma cells in the LP of CD40−/− mice demonstrated few mutations in their Ig V regions, whereas WT LP B cells from different sites showed extensive SHMs, which were also clonally related. Therefore, IgA CSR can occur in PP at a stage preceding manifest GC (GL7-intermediate), whereas SHM require GC formations (GL7-high). These findings reconcile that IgA CSR can occur in PP in the absence of GC with the fact that CD40−/− mice host near normal levels of IgA plasma cells in the LP.
A New Type of Membrane Raft-Like Microdomains and Their Possible Involvement in TCR Signaling
Authors: Otáhal Pavel , Angelisová Pavla , Hrdinka Matouš , Brdička Tomáš , Novák Petr , Drbal Karel , Hořejší Václav
Published: March 2010

Membrane rafts and signaling molecules associated with them are thought to play important roles in immunoreceptor signaling. Rafts differ in their lipid and protein compositions from the rest of the membrane and are relatively resistant to solubilization by Triton X-100 or similar detergents, producing buoyant, detergent-resistant membranes (DRMs) that can be isolated by density gradient ultracentrifugation. One of the key signaling molecules present in T cell DRMs is the transmembrane adaptor protein LAT (linker for activation of T cells). In contrast to previous results, a recent study demonstrated that a LAT construct not present in the buoyant DRMs is fully able to support TCR signaling and development of T cells in vivo. This finding caused doubts about the real physiological role of rafts in TCR signaling. In this study, we demonstrate that these results can be explained by the existence of a novel type of membrane raft-like microdomains, producing upon detergent solubilization “heavy DRMs” containing a number of membrane molecules. At a moderate level of expression, LAT supported TCR signaling more efficiently than constructs targeted to the microdomains producing heavy DRMs or to nonraft membrane. We suggest that different types of membrane microdomains provide environments regulating the functional efficiencies of signaling molecules present therein.
Impaired Lymphoid Organ Development in Mice Lacking the Heparan Sulfate Modifying Enzyme Glucuronyl C5-Epimerase
Authors: Reijmers Rogier M. , Vondenhoff Mark F. R. , Roozendaal Ramon , Kuil Annemieke , Li Jin-Ping , Spaargaren Marcel , Pals Steven T. , Mebius Reina E.
Published: March 2010

The development of lymphoid organs depends on cross talk between hematopoietic cells and mesenchymal stromal cells and on vascularization of the lymphoid primordia. These processes are orchestrated by cytokines, chemokines, and angiogenic factors that require tight spatiotemporal regulation. Heparan sulfate (HS) proteoglycans are molecules designed to specifically bind and regulate the bioactivity of soluble protein ligands. Their binding capacity and specificity are controlled by modification of the HS side chain by HS-modifying enzymes. Although HS proteoglycans have been implicated in the morphogenesis of several organ systems, their role in controlling lymphoid organ development has thus far remained unexplored. In this study, we report that modification of HS by the HS-modifying enzyme glucuronyl C5-epimerase (Glce), which controls HS chain flexibility, is required for proper lymphoid organ development. Glce−/− mice show a strongly reduced size of the fetal spleen as well as a spectrum of defects in thymus and lymph node development, ranging from dislocation to complete absence of the organ anlage. Once established, however, the Glce−/− primordia recruited lymphocytes and developed normal architectural features. Furthermore, Glce−/− lymph node anlagen transplanted into wild-type recipient mice allowed undisturbed lymphocyte maturation. Our results indicate that modification of HS by Glce is required for controlling the activity of molecules that are instructive for early lymphoid tissue morphogenesis but may be dispensable at later developmental stages and for lymphocyte maturation and differentiation.
Anergic CD4+ T Cells Form Mature Immunological Synapses with Enhanced Accumulation of c-Cbl and Cbl-b
Authors: Doherty Melissa , Osborne Douglas G. , Browning Diana L. , Parker David C. , Wetzel Scott A.
Published: March 2010

CD4+ T cell recognition of MHC:peptide complexes in the context of a costimulatory signal results in the large-scale redistribution of molecules at the T cell–APC interface to form the immunological synapse. The immunological synapse is the location of sustained TCR signaling and delivery of a subset of effector functions. T cells activated in the absence of costimulation are rendered anergic and are hyporesponsive when presented with Ag in the presence of optimal costimulation. Several previous studies have looked at aspects of immunological synapses formed by anergic T cells, but it remains unclear whether there are differences in the formation or composition of anergic immunological synapses. In this study, we anergized primary murine CD4+ T cells by incubation of costimulation-deficient, transfected fibroblast APCs. Using a combination of TCR, MHC:peptide, and ICAM-1 staining, we found that anergic T cells make mature immunological synapses with characteristic central and peripheral supramolecular activation cluster domains that were indistinguishable from control synapses. There were small increases in total phosphotyrosine at the anergic synapse along with significant decreases in phosphorylated ERK 1/2 accumulation. Most striking, there was specific accumulation of c-Cbl and Cbl-b to the anergic synapses. Cbl-b, previously shown to be essential in anergy induction, was found in both the central and the peripheral supramolecular activation clusters of the anergic synapse. This Cbl-b (and c-Cbl) accumulation at the anergic synapse may play an important role in anergy maintenance, induction, or both.
Secreted IgM Enhances B Cell Receptor Signaling and Promotes Splenic but Impairs Peritoneal B Cell Survival
Authors: Notley Clare A. , Baker Nicole , Ehrenstein Michael R.
Published: March 2010

B cell survival has a central role in maintaining immune responses to foreign organisms while curbing autoimmunity. In this study, we show that mature B cell survival is impaired and B cell turnover is accelerated in the spleen of mice lacking secreted IgM. Although in vitro responses to BCR cross-linking were normal, there was a marked reduction in basal ERK and global tyrosine phosphorylation in splenic B cells from serum IgM-deficient mice, suggesting diminished interaction with cognate Ag in vivo. The provision of BAFF either in vitro or in vivo reversed the increase in B cell apoptosis, demonstrating that other survival signals can compensate for the loss of secreted IgM in the spleen. In striking contrast to the splenic compartment, peritoneal B cell survival was enhanced in secreted IgM-deficient mice, despite a similar reduction in basal BCR signaling compared with wild type mice. These results suggest that secreted IgM acts as an adjuvant, boosting BCR signals to maintain survival and maintenance of mature splenic B cells while increasing B cell apoptosis in the peritoneum. BAFF administration mitigated the consequences of secreted IgM deficiency on B cell survival in the spleen but not in the peritoneum. This work provides new insight into the regulation of B cell signaling and homeostasis in different peripheral compartments by secreted IgM.
Persistent γ-Herpesvirus Infection Induces a CD4 T Cell Response Containing Functionally Distinct Effector Populations
Authors: Stuller Kathleen A. , Cush Stephanie S. , Flaño Emilio
Published: March 2010

The direct effector mechanisms of CD4 T cells during γ-herpesvirus 68 (γHV68)-persistent infection are less well understood than those of their CD8 T cell counterparts, although there is substantial evidence that CD4 T cells are critical for the control of persistent γ-herpesvirus infection. Our results show that in γHV68-persistently infected mice, CD4 T cells are not cytokine polyfunctional, but there is a division of labor in the CD4 T cell compartment in which CD4 T cells polarize toward two distinct populations with different effector functions: IFN-γ producers and CD107+ cytolytic effectors. These two CD4 T cell effector populations degranulate and produce IFN-γ during steady state without need for exogenous antigenic restimulation, which is fundamentally different from that observed with γHV68-specific CD8 T cells. By using anti–IFN-γ Ab depletions and IFN-γ–deficient mice, we show that CD4 T cell-mediated cytotoxicity in vivo is not dependent on IFN-γ activity. In addition, our data show that purified CD4 T cells isolated from γHV68-latently infected mice have the capacity to inhibit γHV68 reactivation from latency. Our results support the concept that CD4 T cells are critical effectors for the control of γ-herpesvirus latent infection, and they mediate this effect by two independent mechanisms: IFN-γ production and cytotoxicity.
Coordinate Interaction between IL-13 and Epithelial Differentiation Cluster Genes in Eosinophilic Esophagitis
Authors: Blanchard Carine , Stucke Emily M. , Burwinkel Karen , Caldwell Julie M. , Collins Margaret H. , Ahrens Annette , Buckmeier Bridget K. , Jameson Sean C. , Greenberg Allison , Kaul Ajay , Franciosi James P. , Kushner Jonathan P. , Martin Lisa J. , Putnam Philip E. , Abonia J. Pablo , Wells Suzanne I. , Rothenberg Marc E.
Published: March 2010

We have previously proposed that the pathogenesis of eosinophilic esophagitis (EE) is mediated by an IL-13–driven epithelial cell response associated with marked gene dysregulation including eotaxin-3 overproduction. In this study, we compared epithelial responses between healthy patients and those with EE, aiming to uncover molecular explanations for EE pathogenesis. Esophageal epithelial cells could be maintained for up to five passages, with 67% and 62% of cell lines reaching confluence in healthy controls and EE cases, respectively. Both sets of epithelial cells avidly responded to IL-13 at similar levels as assessed by eotaxin-3 production. Acidic pH increased cellular release of eotaxin-3 (4.6 ± 1.98 ng/ml versus 12.46 ± 2.90 ng/ml at pH 7.4 and 4, respectively; p < 0.05). Numerous epidermal differentiation complex (EDC) genes, such as filaggrin and SPRR3, were downregulated both in IL-13–stimulated esophageal epithelial cells and in EE biopsies specimens compared with healthy controls. Whereas the filaggrin loss of function mutation 2282del4 was overrepresented in EE compared with control individuals (6.1% versus 1.3% respectively; p = 0.0172), the decreased filaggrin expression was uniformly seen in all EE cases in vivo. Indeed, expression of the EDC genes filaggrin and involucrin was strongly decreased directly by IL-13. These results establish that the epithelial response in EE involves a cooperative interaction between IL-13 and expression of EDC genes.