The clinicopathological feature of primary cutaneous lymphomas according to WHO/EORTC classification and their relationship to EBV in Taiwan has never been reported. This retrospective study collected the patients with cutaneous lymphomas from 1990 and 2006. The morphology, EBER in situ hybridization and immunohistochemistry of primary cutaneous lymphomas were studied to reclassify based on the WHO/EORTC classification. A total of 54 patients were included. Twenty-nine were primary cutaneous lymphomas and 25 were secondary cutaneous lymphomas. The age ranged from 21 to 86 years old (mean 62 years old). Twenty-one (72.4%) were primary cutaneous T-cell and NK-cell lymphoma, including 5 extranodal NK/T-cell lymphoma, nasal type (17.2%), 5 primary cutaneous peripheral T-cell lymphoma, unspecified (17.2%), 4 mycosis fungoides (13.8%), 1 Sezary syndrome, 3 primary cutaneous anaplastic large cell lymphoma, 2 primary cutaneous small-medium CD4+ T-cell lymphoma and 1 subcutaneous panniculitis-like T-cell lymphoma. Eight cases were primary cutaneous B-cell lymphoma (27.6%) including 3 cutaneous marginal zone B-cell lymphoma (10.3%), 3 cutaneous follicle center B-cell lymphoma (10.3%), and 2 diffuse large B-cell lymphoma, leg type (6.9%). Seventeen cases were secondary cutaneous T-cell and NK -cell lymphoma. Eight cases were secondary cutaneous B-cell lymphoma. All primary and secondary extranodal NK/T-cell lymphoma, nasal type, were positive for EBER, however, one of them (10%) without both angiocentric growth pattern and necrosis in histomorphological examination. This is the first clinicopathological study of cutaneous lymphoma according to recent WHO/EORTC classification in Taiwan. In comparison with the Western countries, mycosis fungoides is less common whereas primary extranodal NK/T-cell lymphoma, nasal type, and peripheral T-cell lymphoma, unspecified, is more common in Taiwan. EBER in situ hybridization study is helpful in the diagnosis of extranodal NK/T-cell lymphoma, nasal type, especially in tumor without both angiocentric growth pattern and necrosis.

Thèse de doctorat : Aspects moléculaires et cellulaires de la biologie : Orléans : 2009. Thèse de doctorat : Aspects moléculaires et cellulaires de la biologie : Académie des sciences de Pologne : 2009. / Thèse soutenue en co-tutelle. Titre provenant de l'écran-titre.

The NK cell is a large granular lymphocyte that plays a key role in protecting the body against numerous pathogens including parasites, intracellular bacteria, viral infections, as well as showing anti-tumor activity and playing a role in the rejection of allogeneic BM. Unlike other lymphocytic cell types, that utilize rearranging receptors, NK cells are regulated by a complex array of germ line encoded activating and inhibitory receptors. NK cells are often described as a front line or rapid defense given their response to stimuli can be immediate, although they also maintain functions that extend their role well into the adaptive immune system. Inhibitory receptors that recognize MHC class I molecules regulate NK cell responses and self-tolerance. Recent evidence indicates self-ligands not present in the MHC locus can also modulate NK function. We previously demonstrated that the NK receptor repertoire is disrupted by SHIP-deficiency. Here we show that an inhibitory receptor, 2B4, that recognizes an MHC-independent ligand is over expressed in NK cells of SHIP-/- mice at all stages of NK development and differentiation. Overexpression of 2B4 compromises key cytolytic NK functions, including killing of allogeneic, tumor and viral targets. These results demonstrate that in SHIP-/- NK cell 2B4 is the dominant inhibitory receptor. We then furthered this finding by examining the molecular basis of 2B4 dominance. We show that in SHIP-/- NK cells there is increased 2B4 expression as well as a strong bias towards the 2B4L isoform. We have also identified a greater than tenfold increase in SHP1 recruitment to 2B4. Consistent with this SHP1 over recruitment,both a broad and a selective SHP1 inhibitor restore SHIP-/- NK killing of complex targets.Through this study we have identified the molecular mechanism of 2B4 receptor dominance as SHP1 over-recruitment.In addition we have utilized protein array technology to explore NK signaling through the determination of the NK kinome. To this end we have been able to identify multiple pathways that may mark crucial differences between the mature and immature NK cell.

This thesis investigates the manufacture and use of ceramics over four centuries in Lincolnshire, and considers the evidence for date and function of the pottery itself and for the social standing and economy of the potters, late survivors of the medieval peasant craftsman tradition. Documentary and physical evidence are both searched to produce the most comprehensive possible list of sites and potters names, and to highlight the areas of doubt where neither type of source can give sufficient proof. The methods of pottery production are also examined and two specific types of vessels are discussed in detail as examples of the search for -=origins. From this point the search widens to consider the importation principally by sea of pottery from other parts of the country and from Europe, and the prices of such wares are compared with prices of local products. This leads to certain conclusions about the economic pressures on local potters and their adjustments to deal with new problems and changing expectations. Contemporary sources, depositional evidence and context are next used to study the names and function of pottery, and finally the principles of dating are discussed, and a series of pottery groups are analysed to test the reliability and transferability of dating. Throughout pottery making is compared with comparable trades and Lincolnshire's position with that of the wider ceramic world.

Ancient glasses are invariably complex materials, in which the specific chemical composition and microstructure capture aspects of their technologies. The chemical characterisation of glasses in specific archaeological contexts has given useful insight into the peculiarities of diverse glass-making technologies. In addition such studies generate more general information upon an important range of phenomenon, including the pyrotechnological milieu, empirical knowledge of sophisticated chemistry, organisation of production, access to significant raw materials and long-distance trade. This study examines a wide selection of glass artefacts recovered from archaeological contexts in Northern and Central Italy from approximately 1200 BC to 200 BC. The earliest material is from the Final Bronze Age, and extends the characterisation of an established glass type, which is unique to Europe and distinct from the contemporary technologies of the Eastern Mediterranean. Using a combination of X-ray fluorescence analysis, electron microprobe and scanning electron microscopy glass artefacts from a thousand-year period from the same region are investigated. The shifting technologies permit the discussion of localised production and importation of glass from elsewhere. The chemical analysis reveals a complex picture of glass production, which defies the expected pattern, and there is evidence for new compositional types, which may yet prove to be diagnostic of highly localised production. The changing compositions are discussed in relation to the broader archaeological context.

We comparatively analyzed the natural killer (NK) cell response against HCMV-infected pro-inflammatory (M1) and anti-inflammatory (M2) M[Fi] derived from autologous monocytes. M1 M[Fi] were more resistant to infection, secreting TNF-[alfa], IL-6, IL-12 and type I IFN. By contrast, in HCMV-infected M2 M[Fi] the production of proinflammatory cytokines, type I IFN and IL-10 was limited, and IL-12 undetectable. NK cell degranulation was triggered by interaction with HCMV-infected M1 and M2 M[Fi] and was partially inhibited by specific anti-NKp46, anti-DNAM-1 and anti-2B4 mAbs, thus supporting a dominant role of these activating receptors. By contrast, only HCMV-infected M1 M[Fi] efficiently promoted NK cell-mediated IFN-[gamma] secretion, an effect partially related to IL-12 production. These observations reveal differences in the NK cell response triggered by distinct HCMV-infected monocyte-derived cell types, which may be relevant in the pathogenesis of this viral infection. HCMV infection has been proposed to contribute to the development of atherosclerosis, a chronic inflammatory process in which M[Fi] play a key role. The contribution of HCMV to vascular disease may depend on features of the immune response not reflected by the detection of specific antibodies. Persistent HCMV infection in healthy blood donors has been associated with changes in the distribution of NK cell receptors (NKR). The putative relationship among HCMV infection, NKR distribution, subclinical atherosclerosis and coronary heart disease was assessed. An association of overt and subclinical atherosclerotic disease with LILRB1+ NK and T cells was observed, likely reflecting a relationship between the immune challenge by infections and cardiovascular disease risk, without attributing a dominant role for HCMV. / Hem analitzat la resposta de la cèl•lula NK als macròfags proinflamatoris (M1) i antiinflamatoris (M2) derivats de monòcits autòlegs infectats pel citomegalovirus humà (HCMV). Els macròfags M1 son més reistents a la infecció i secreten TNF-[alfa], IL-6, IL-12 i IFN de tipus I. Per altra banda, en els macròfags M2 infectats per HCMV la producció de citoquines proinflamatories, IFN de tipus I i IL-10 es limitada i la IL-12 indetectable. La cèl•lula NK degranula al interaccionar amb els macròfags M1 i M2 infectats. Aquesta degranulació s’inhibeix parcialment al bloquejar amb anticossos específics anti-NKp46, anti-DNAM-1 i anti-2B4, això indica que aquests receptors tenen un rol important en el procés. En canvi, només els macròfags M1 infectats amb HCMV promouen de manera eficient la producció d’IFN-[gamma] per part de la cèl•lula NK, degut parcialment a la producció de IL-12. Aquestes observacions posen de manifest diferències en la resposta de la cèl•lula NK a diferents tipus de macròfags infectats per HCMV que pot ser relevant en la patogènesis d’aquesta infecció viral. S’ha proposat que la infecció per HCMV contribueix al desenvolupament de l’aterosclerosis, un procés inflamatori crònic en el que els macròfags tenen un paper clau. La contribució del HCMV a la malaltia cardiovascular pot dependre de la resposta immune. La infecció per HCMV en donants de sang sans s’ha associat a canvis en la distribució dels receptors de les cèl•lules NK. S’ha evaluat la possible relació entre la infecció per HCMV, la distribució dels receptors de les cèl•lules NK i l’infart agut de miocardi. S’ha observat una associació de l’infart agut de miocardi i l’aterosclerosi subclínica tant amb les cèl•lules NK LILRB1+ com amb les cèl•lules T LILRB1+. Això possiblement reflexa la relació entre la pressió que les infeccions exerceixen en el sistema immunitari i el risc cardiovascular sense atribuir un paper principal al HCMV.

The immune system is comprised of specific cell types that communicate and interact via a range of soluble and surface-bound molecules to defend the body against pathogens. Many gaps remain in our understanding of the subcellular trafficking pathways that regulate the diverse functions of the immune system. The central aim of this thesis was to investigate transport through the endocytic pathway, focussing in particular on the unique organelles and functions of this pathway in immune cells. Two subsets of immune cells were of particular interest in this thesis, macrophages and natural killer (NK) cells. These cell types both perform a range of functions that contribute to both innate and adaptive immunity. Another common thread between these cells is that they both perform functions involving specialised endocytic organelles and pathways. Macrophages utilise their endocytic pathways to perform several unique functions; phagocytosis, endocytosis and degradation of foreign proteins for presentation on MHC class II molecules, and signalling of Toll-like receptors from endosomes. Even secretion of cytokines such as tumour necrosis factor alpha (TNFα) by macrophages requires transport through an endosomal compartment, the recycling endosome, as recently discovered in this laboratory (Murray et al., 2005a). NK cells utilise specialised secretory lysosomes to deliver a lethal hit to carefully identified target cells, providing an alternative example of specialised endocytic trafficking in the immune system. Of the many protein families that regulate subcellular trafficking, the SNARE, Rab, Munc and exocyst proteins were focussed on during this thesis. The localisation and function of members of these families in the endocytic pathway were investigated. Novel results in macrophages concerned the role of Vti1b in endocytosis, a process with implications for MHC class II antigen presentation and TLR detection of endocytosed particles. Alteration of Vti1b protein levels in the cells significantly decreased uptake and degradation of endocytic cargo. A role for Rab11 and the recycling endosome in antigen presentation was also studied. MHC class II was detected in recycling endosomes, and overexpression of a mutant Rab11 protein altered the distribution of MHC class II, suggesting a role for Rab11 in subcellular trafficking during antigen presentation. Preliminary results also suggest a role for the exocyst protein Sec15 at the recycling endosome in macrophages, providing a new target for investigation of the regulation of TNFα secretion. The recycling endosome is emerging as a vital transport hub during cytokine secretion, phagocytosis and possibly other cellular functions in macrophages. This project also involved the unique opportunity to examine primary NK cells from patients with a number of genetic immunodeficiencies caused by mutations to trafficking proteins. The autosomal recessive immunodeficiencies Griscelli syndrome type 2 (GS2) and familial haemophagocytic lymphohistiocytosis types 3 (FHL3) and 4 (FHL4) are associated with loss-of-function of Rab27a, Munc13-4 and syntaxin 11 (Stx11), respectively. These diseases involve a loss of cytotoxic function by cytotoxic CD8+ T lymphocytes and NK cells, but the precise molecular role of these proteins in granule release is incompletely understood. In freshly isolated, resting NK cells from healthy subjects, PMA and ionomycin stimulation or conjugation to susceptible target cells induced colocalisation of endogenous Rab27a and Munc13-4 to perforin-containing granules. In Rab27a-deficient cells, which showed defective degranulation and cytotoxicity induced by signals for both natural and antibody-dependent cellular cytotoxicity, Munc13-4 failed to colocalise with perforin upon activation. Unexpectedly, Rab27a and Munc13-4 localisation to lytic granules was selectively induced by different receptor signals, demonstrating specificity for regulation of lytic granule maturation by target cell ligand expression. Recruitment of the SNARE protein Vti1b, which has not previously been associated with NK cell function or secretory lysosome release, to perforin granules was also discovered. Unexpectedly, Stx11 was not localised to perforin granules. These experiments have contributed to our understanding of the precise molecular roles of Munc13-4, Rab27a and Stx11 in NK cell granule release. Overall, this thesis presents novel and important results from studies of subcellular transport through the endocytic pathways of macrophages and NK cells. These results advance our understanding of several immune functions, and a number of human genetic immunodeficiencies. This new knowledge of the role of endocytic organelles and fusion machinery in these processes provides exciting targets for future research.

Thèse (M.Sc.) -- Université Laval, 1997. / Bibliogr.: f. 69-74. Publié aussi en version électronique.

Nous avons montré que, dans l'intestin, les cellules exprimant le marqueur spécifique des lymphocytes Natural Killer (NK), NKp46 peuvent être divisées en deux compartiments, par rapport au facteur de transcription RORyt. Les cellules NKp46+RORyt- sont des cellules NK intestinales immatures. Cependant, les cellules NKp46+RORyt- s'apparentent aux cellules "lymphoid tissue-inducer" (LTi), puisqu'elles requièrent RORyt pour leur développement et produisent de l'IL-22, cytokine essentielle pour l'homéostasie et la réponse antimicrobienne épithéliale. Notre recherche actuelle vise à établir les relations ontogéniques et la signature génétique de ces cellules au niveau pan-génomique. La fonction in vivo des cellules NKp46+ intestinales est adressée dans un modèle unique d'infection par voie orale avec Listeria monocytogenes. L'ensemble de nos résultats a montré que les cellules NKp46+ ont une place légitime dans la famille émergente des cellules intestinales de l'immunité innée. / Natural Killer (NK) cells are NKp46+CD3- innate lymphocytes, which exhibit cytotoxicityand cytokine production, mainly IFN-γ, as major effector functions. In mammals, theexpression of natural cytotoxic receptor NKp46 is essentially restricted to NK cellcompartment. However, we showed that in mouse intestine, NKp46 marker defines aheterogeneous cell population, differentially expressing the retinoic acid orphan receptorRORγt. NKp46+RORγt- cells harbor reduced cytotoxicity and IFN-γ production, consistentwith an immature NK cell phenotype. In contrast, NKp46+RORγt+ cells resemble lymphoidtissue inducer cells (LTi) in their developmental requirement for RORγt and their ability toproduce IL-22. This cytokine is critical for epithelial homeostasis and antimicrobial responseat several epithelial sites.By using a genome wide profiling approach, we confirmed that gut NKp46+RORγt- cellsare the only gut NKp46+ cell population related to conventional splenic NK cells, while gutNKp46+RORγt+ cells are linked to LTi cells. Transcriptional signatures specific of distinct gutNKp46+ cell subsets are currently under intensive investigation in order to determine novelfunctional properties.We assessed gut NKp46+ cell subsets in vivo function during oral infection with entericpathogen Listeria monocytogenes (L.m.). Although immune responses to systemic L.minfection have been widely characterized in mice, L.m. cannot breach mouse intestinal barrier,thus limiting the knowledge of early immune responses at the natural site of infection. Using aunique transgenic mouse lineage restoring L.m. intestinal invasiveness, we showed that gutNKp46+RORγt- and NKp46+RORγ t+ cells respond to L.m. infection by producing IFN-γ andIL-22, respectively. Further investigation of the cellular mechanisms leading to gut NKp46+cell subset activation, as well as the contribution of these cells to the control of bacterialdissemination is in progress.Altogether our data provide novel insight into intestinal innate immunity and highlight gutNKp46+ cell subsets as “legitimate” members of the emerging family of intestinal innatelymphoid cells

Submitted by Hiolanda Rêgo (hiolandarego@gmail.com) on 2017-03-10T14:19:47Z No. of bitstreams: 1 Tese_Med_ Taís Campos.PDF: 1941952 bytes, checksum: c97f06b80eaaa998c41ad42a9d48dfc7 (MD5) / Approved for entry into archive by Delba Rosa (delba@ufba.br) on 2017-06-28T12:16:30Z (GMT) No. of bitstreams: 1 Tese_Med_ Taís Campos.PDF: 1941952 bytes, checksum: c97f06b80eaaa998c41ad42a9d48dfc7 (MD5) / Made available in DSpace on 2017-06-28T12:16:30Z (GMT). No. of bitstreams: 1 Tese_Med_ Taís Campos.PDF: 1941952 bytes, checksum: c97f06b80eaaa998c41ad42a9d48dfc7 (MD5) / Introdução: Leishmaniose cutânea (LC) devido à infecção por L. braziliensis é caracterizada pelo desenvolvimento de lesões cutâneas ulcerativas. Recentemente, atenção tem sido dada à participação das células T CD8+ para a resposta inflamatória deletéria observada em pacientes com LC. Foi demonstrado que estas células contribuem para a imunopatologia através da produção de citocinas pró-inflamatórias, granzima e perforina. Em adição às células T CD8+ , células NK e NKT são também importantes fontes de citocinas pró-inflamatórias, granzima e perforina, e o papel destas células na infecção por L. braziliensis ainda não foi investigado. Objetivo: Avaliar o papel das células NK na infecção por L. braziliensis. Métodos: As células mononucleares de sangue periférico foram obtidas a partir de indivíduos saudáveis e de pacientes com LC para a marcação ex-vivo com CD56, CD8, CD3, e marcação intracelular para granzima e perforina. As células derivadas de biópsias foram obtidas para munofenotipagem por citometria de fluxo e imunohistoqímica. Resultados: Identificamos quatro sub-populações de células citotóxicas: células NK (CD8- CD3- ), células NK (CD8dimCD3- ), células NKT-like (CD8brigth CD3+ ) e células T CD8+ . Observamos que as frequências de sub-populações de células NK (CD8- e CD8+ ) estavam aumentadas no sangue periférico de pacientes com LC quando comparadas com IS, e estas duas populações expressam mais granzima e perforina do que as células T CD8+ em pacientes com LC. Observamos ainda que as células NK derivadas de lesões de pacientes com LC expressam mais CD107a, marcador de degranulação, do que as células T CD8+ . E que as células NK contribuem para destruição de parasitos na infecção por L. braziliensis Conclusão: Os nossos dados sugerem uma dicotomia funcional das células NK na leishmaniose cutânea, participando tanto na patogênese da doença quanto na destruição de parasitos. Palavras-chave: leishmaniose cutânea, células NK, infócitos citotóxicos, citotoxicidade.