Role of the naive and memory CD4+T-cell repertoire in transplantation

Purpose: The exact role of individual T cell-subsets in the development of rejection is not clearly defined. Given their distinct phenotypes, effector functions and trafficking patterns, naïve (CD45RBhiCD44lo) and memory (CD45RBloCD44hi) T cells may play distinct roles in anti-donor immunity after transplantation. Furthermore, only the CD4+CD45RBlo population contains CD4+CD25+ T cells, a subset with suppressive functions playing a major role in the maintenance of peripheral tolerance. The aim of this work was to study the contribution of these individual subsets in alloresponses via the direct and indirect pathways using a murine experimental model. Methods and materials: Purified naïve or memory CD4+ T cells were adoptively transferred into lymphopenic mice undergoing a skin allograft. Donor to recipient MHC combinations were chosen in order to study the direct and the indirect pathways of allorecognition separately. Graft survival and in vivo expansion, effector function and trafficking of the transferred T cells was assessed at different time points after transplantation. Results: We found that the cross-reactive CD4+CD45RBlo memory T-cell pool was heterogeneous and contained cells with regulatory potentials, both in the CD4+CD25+ and CD4+CD25-populations. CD4+ T cells capable of inducing strong primary alloreactive responses in vitro and rejection of a first allograft in vivo were mainly contained within the CD45RBhi naïve CD4+ T-cell compartment. CD4+CD45RBlo T cells proliferated less abundantly to allogeneic stimulation than their naïve counterparts both in vitro and in vivo, and allowed prolonged allograft survival even after the depletion of the CD4+CD25+ subset. Interestingly, CD4+CD25-CD45RBlo T cells were capable of prolonging allograft survival, mainly when the indirect pathway was the only mechanism of allorecognition. The indirect pathway response, which was shown to drive true chronic rejection and contribute to chronic allograft dysfunction, was predominantly mediated by naïve CD4+ T cells. Conclusion: This work provides new insights into the mechanisms that drive allograft rejection and should help develop new clinical immunosuppressive protocols. In particular, our results highlight the importance of selectively targeting individual T-cell subsets to prevent graft rejection but at the same time maintain immune protective responses to common pathogens.

T cell receptor V beta repertoire in mice lacking endogenous mouse mammary tumor provirus.

When endogenous mouse mammary tumor virus (MMTV) superantigens (SAg) are expressed in the first weeks of life an efficient thymic deletion of T cells expressing MMTV SAg-reactive T cell receptor (TcR) V beta segments is observed. As most inbred mouse strains and wild mice contain integrated MMTV DNA, knowing the precise extent of MMTV influence on T cell development is required in order to study T cell immunobiology in the mouse. In this report, backcross breeding between BALB.D2 (Mtv-6, -7, -8 and -9) and 38CH (Mtv-) mice was carried out to obtain animals either lacking endogenous MMTV or containing a single MMTV locus, i.e. Mtv-6, -7, -8 or -9. The TcR V beta chain (TcR V beta) usage in these mice was analyzed using monoclonal antibodies specific for TcR V beta 2, V beta 3, V beta 4, V beta 5, V beta 6, V beta 7, V beta 8, V beta 11, V beta 12 and V beta 14 segments. Both Mtv-8+ mice and Mtv-9+ mice deleted TcR V beta 5+ and V beta 11+ T cells. Moreover, we also observed the deletion of TcR V beta 12+ cells by Mtv-8 and Mtv-9 products. Mtv-6+ and Mtv-7+ animals deleted TcR V beta 3+ and V beta 5+ cells, and TcR V beta 6+, V beta 7+ and V beta 8.1+ cells, respectively. Unexpectedly, TcR V beta 8.2+ cells were also deleted in some backcross mice expressing Mtv-7. TcR V beta 8.2 reactivity to Mtv-7 was shown to be brought by the 38CH strain and to result from an amino acid substitution (Asn-->Asp) in position 19 on the TcR V beta 8.2 fragment. Reactivities of BALB.D2 TcR V beta 8.2 and 38CH TcR V beta 8.2 to the exogenous infectious viruses, MMTV(SW) and MMTV(SHN), were compared. Finally, the observation of increased frequencies of TcR V beta 2+, V beta 4+ and V beta 8+ CD4+ T cell subsets in Mtv-8+ and Mtv-9+ mice, and TcR V beta 4+ CD4+ T cells in Mtv-6+ and Mtv-7+ mice, when compared with the T cell repertoire of Mtv- mice, is consistent with the possibility that MMTV products contribute to positive selection of T cells.

An allele-specific, stochastic gene expression process controls the expression of multiple Ly49 family genes and generates a diverse, MHC-specific NK cell receptor repertoire.

Mouse NK cells express MHC class I-specific inhibitory Ly49 receptors. Since these receptors display distinct ligand specificities and are clonally distributed, their expression generates a diverse NK cell receptor repertoire specific for MHC class I molecules. We have previously found that the Dd (or Dk)-specific Ly49A receptor is usually expressed from a single allele. However, a small fraction of short-term NK cell clones expressed both Ly49A alleles, suggesting that the two Ly49A alleles are independently and randomly expressed. Here we show that the genes for two additional Ly49 receptors (Ly49C and Ly49G2) are also expressed in a (predominantly) mono-allelic fashion. Since single NK cells can co-express multiple Ly49 receptors, we also investigated whether mono-allelic expression from within the tightly linked Ly49 gene cluster is coordinate or independent. Our clonal analysis suggests that the expression of alleles of distinct Ly49 genes is not coordinate. Thus Ly49 alleles are apparently independently and randomly chosen for stable expression, a process that directly restricts the number of Ly49 receptors expressed per single NK cell. We propose that the Ly49 receptor repertoire specific for MHC class I is generated by an allele-specific, stochastic gene expression process that acts on the entire Ly49 gene cluster.

Anti-HLA antibody repertoire after IVIg infusion in highly sensitized patients waiting for kidney transplantation.

Polyclonal intravenous immunoglobulin (IVIg) treatment reduces crossmatch positivity and increases rates of transplantation in highly sensitised patients (HS). We quantified the panel reactive antibody (PRA) by microlymphocytotoxicity (MLCC), and we analysed anti-HLA class I and class II IgG specific antibody repertoire by Luminex before and after IVIg infusion alone in HS patients awaiting kidney transplantation. Five patients received three monthly infusions of 1 g/kg of IVIg. Serum samples collected pre and post IVIg treatment were submitted for PRA analysis by MLCC. Anti-class I and anti-class II antibody specificities were then tested by Luminex. We focused on the anti-HLA class I and class II antibodies directed against HLA expressed by a previous graft. We also analysed the anti-HLA antibody repertoire in three patients who had not received IVIg infusion. The PRA level determined by MLCC decreased significantly in one of the five patients, dropping from 40% to 17%. The Luminex assay showed fluctuations of the anti-HLA antibody levels over time, but no significant longterm modifications of the anti-HLA antibody repertoire were observed, even in the patient with a strong and prolonged reduction of the PRA determined by MLCC. Our results show that IVIg at 1 g/kg is not sufficient to reduce PRA and does not modify the repertoire of specific anti-HLA antibody determined by Luminex.

Les étudiant(e)s en médecine mènent une recherche dans La communauté [Male/female medical students conduct research in the community].

Pendant quatre semaines, les étudiant(e)s en médecine de 3e année de l'Université de Lausanne mènent une enquête dans la communauté sur le sujet de leur choix. L'objectif de ce module est de faire découvrir aux futurs médecins les déterminants non biomédicaux de la santé, de la maladie et de l'exercice de la médecine : les styles de vie, les facteurs psychosociaux et culturels, l'environnement, les décisions politiques, les contraintes économiques, les questions éthiques, etc. Par groupes de cinq, les étudiant(e)s commencent par définir une question de recherche originale et en explorent la littérature scientifique. Leur travail de recherche les amène à entrer en contact avec le réseau d'acteurs de la communauté concernés, professionnels ou associations de patients dont ils analysent les rôles et influences respectives. Chaque groupe est accompagné par un(e) tuteur(trice), enseignant(e) de la Faculté de biologie et de médecine de l'Université de Lausanne. Les étudiant(e)s présentent la synthèse de leurs travaux pendant un congrès de deux jours à la fin du module. Quatre travaux parmi les plus remarquables ont été choisis pour être publiés dans la Revue Médicale Suisse et Primary Care.

Soldiers in a stingless bee: work rate and task repertoire suggest they are an elite force

The differentiation of workers into morphological subcastes (e.g., soldiers) represents an important evolutionary transition and is thought to improve division of labor in social insects. Soldiers occur in many ant and termite species, where they make up a small proportion of the workforce. A common assumption of worker caste evolution is that soldiers are behavioral specialists. Here, we report the first test of the "rare specialist" hypothesis in a eusocial bee. Colonies of the stingless bee Tetragonisca angustula are defended by a small group of morphologically differentiated soldiers. Contrary to the rare specialist hypothesis, we found that soldiers worked more (+34%-41%) and performed a greater variety of tasks (+23%-34%) than other workers, particularly early in life. Our results suggest a "rare elite" function of soldiers in T. angustula, that is, that they perform a disproportionately large amount of the work. Division of labor was based on a combination of temporal and physical castes, but soldiers transitioned faster from one task to the next. We discuss why the rare specialist assumption might not hold in species with a moderate degree of worker differentiation.

The Impact of Supervisor's Advice, Codes of Conduct, Accountability and Moral Attentiveness of Individuals on Discriminatory Behaviour Against Immigrants.

Past studies on the personnel selection demonstrated that a supervisor's advice to discriminate can lead to compliant behaviours. This study had the aim to extend past findings by examining what can overcome the powerful influence of the hierarchy. 50 Swiss managers participated to an in-basket exercise. The main task was to evaluate Swiss candidates (in-group) and foreigners (out-groups: Spanish and Kosovo Albanians) and to select two applicants for a job interview. Main results were the effect of codes of conduct to prevent discrimination against out-group applicants in the presence of a supervisor's advice to prefer in-group members. But, when participants were accountable to an audience, this beneficial effect disappears because participants followed the supervisor's advice. The second aim was to assess if the difference in responses between participants was related to their difference in moral attentiveness. Results showed some significant relationships but not always in the direction expected.

A proposal for a repertoire of software testing methods

Nowadays software testing and quality assurance have a great value in software development process. Software testing does not mean a concrete discipline, it is the process of validation and verification that starts from the idea of future product and finishes at the end of product’s maintenance. The importance of software testing methods and tools that can be applied on different testing phases is highly stressed in industry. The initial objectives for this thesis were to provide a sufficient literature review on different testing phases and for each of the phases define the method that can be effectively used for improving software’s quality. Software testing phases, chosen for study are: unit testing, integration testing, functional testing, system testing, acceptance testing and usability testing. The research showed that there are many software testing methods that can be applied at different phases and in the most of the cases the choice of the method should be done depending on software type and its specification. In the thesis the problem, concerned to each of the phases was identified; the method that can help in eliminating this problem was suggested and particularly described.

Restaurant Trocadero : Repertoire

Orkesterin soitto-ohjelmisto.

Musikalisk lärandemiljö : planerade musikaktiviteter med småbarn i daghem

A musically stimulating nvironment is of great significance when considering the opportunities for development and learning that are offered to children in day nurseries. Moreover, the quality of musical activities also has great significance for the musical development of growing children. Young children are always striving to create meaning and understanding; they influence and are influenced by the context in which they are situated. The purpose of this study is to investigate the planned music activities in the musical learning environment of seven toddler’s groups in day nurseries, based on four different aspects: the texts of children’s songs, the melodies of children’s songs, the basic elements of music and the use of instrumentation. The data has been gathered by video observation of the planned music activities. The age of the children studied ranged from 11 months to 3 years and 11 months, and on average there were 13 children in each group. The study involved a total of just over 90 children and nine educators. The methodological approaches are hermeneutic as well as videographic, and they are applied to the analysis as required. The approaches are well suited to an understanding of the planned music activities where the individual’s communications and actions are studied. The results of the study show that educators impart a cultural heritage to the children in the form of old traditional songs, but they also act as intermediaries for newer children's songs. They focus on actively on the text content of the children’s songs in their interaction with the children, but do not address the meaning of the text. Furthermore, while the educators use music sessions for the children to develop, among other things, an understanding of language and social skills, they do not use the sessions to develop the children’s musical skills. The educators could, to a much greater extent than they now do, direct the children's attention to the basic elements of music, and conduct meta-cognitive dialogues with the children. They use a large repertoire of children's songs, which are, however, often too demanding for the children’s voices to cope with. In addition, the educators do not plan, in any significant way, activities for the toddlers in order to help them develop the accuracy of their vocal tones. With regard to the use of instruments, the educators focus, in their interaction with the children, is placed on both the usage of the instruments and knowledge about the instruments. Regarding the use of instruments the study shows that the children’s musical expression takes place in an environment that includes both melody and rhythm instruments. It is clear that children are actively interested in the constructed instrument environment, because for long periods of time they independently play and experiment with instruments.

Cafe- & Restaurant Cosmopolite : Repertoire

Orkesterin ohjelmisto. Kertosäkeistöjen sanoja yleisön laulettavaksi.

Supplier codes of conduct: Acting responsibly in global supply chains

Nowadays the Western companies are considered responsible for the social and environmental issues in their whole supply chains. To influence the practices of their suppliers the Western companies have created suppliers codes of conduct (SCCs) which express their requirements. Suppliers’ compliance with the SCCs is checked through audits. The purpose of this thesis is to analyze SCCs as a means for Western companies to ensure socially and environmentally responsible actions in their global supply chains, and the sub-objectives are to find out 1) how well do the SCCs and their auditing work at suppliers’ production sites and 2) how can possible problems related to SCCs and their auditing be solved. This is a qualitative research carried out in the form of a case study with two case companies. In this study both primary and secondary data is used. The primary data is collected in the form of interviews of the case company representatives and three external experts. Based on a theoretical framework of previous research in the fields of corporate social responsibility and supply chain management, a model with eleven factors, which influence the success of SCC implementation and the auditing of SCC –implementation, is drafted. Also several different best-practices to help to solve and avoid possible problems related to SCC -implementation and auditing have been identified from previous research. Based on the findings of this study the theoretical model has been updated adding two new influential factors. It seems that how well the SCC and its auditing work at suppliers’ production sites depends on the joint effect of thirteen influential factors: buyer’s purchasing policy, supplier’s motivation, buyer’s commitment, the solving of agency problems, the contents of the SCC, supplier’s role and the buyer-supplier –relationship, complexity of supply chain, the limitations of the smaller buyers, cooperation through a business association or multi-stakeholder system, the role of supplier’s employees, SCC –related communication and supplier’s understanding, cheating in audits and the auditors. The possible problems related to SCCs and their auditing can be solved by adopting best-practices. Nine of the theoretical best-practices stand out from the findings of this study: 1) two-way communication and collecting feedback from suppliers, 2) the philosophy of continuous improvement, 3) long-term business relationships with the supplier, 4) informing the supplier about the advantages of SCC –compliance, 5) rewarding code-compliant suppliers, 6) building collaborative, good buyer-supplier relationships, 7) supporting and advising the supplier, 8) joining a business association or multi-stakeholder system and 9) interviewing supplier’s employees as a part of the audits.

Reinventing the third way : towards new forms of economic and democratic conduct

Avhandlingen studerar samspelet mellan det politiska och det ekonomiska systemet genom det föregående seklet, med speciellt fokus på den nyliberala omdaning som utspelats de senaste tre årtiondena. Med en liberaliserad ekonomi och en individualiserad människa måste också det politiska systemet utformas på ett annat sätt. Gradvisa förändringar har varit synliga, som den växande nätverksorganiseringen, dvs. att olika typer av utvecklingspolitik allt mer genomförs som ett samspel mellan samhällssektorerna, istället för att enbart utföras av den offentliga sektorn som tidigare. Genom fyra fallstudier i Österbotten demonstreras hur arenor för samverkan kan konstrueras i enlighet med vad den aktuella frågan kräver. Ett exempel är det innovationsnätverk som skapats i Karleby-Jakobstad-regionen, där offentliga och privata intressenter deltar, och som samtidigt bryter traditionella administrativa gränser. Ett annat mer negativt exempel är omstruktureringsprocessen i Kaskö-regionen eft er Metsä-Botnia-fabrikens nedläggning, där småkommunproblemet tillsammans med att den privata intressenten inte haft skyldighet att samverka lett till att en tillräckligt regionalt förankrad arena för en långsiktig utvecklingspolitik inte har kunnat skapas. Det här visar på trögheten i det finska systemet, där en stark enhetsstat och en splittrad kommunal nivå ur ett historiskt perspektiv hindrat flexibla anpassningar till samhällsförändringar. Avhandlingen visar att nätverksorganiseringen bör vara en integrerad del av de representativa strukturerna för att kunna påverka processernas utfall. Avhandlingen visar framförallt att deltagande och funktionellt definierade regionala och lokala arenor har stor potential, både ekonomiskt och demokratiskt. Det handlar om att skapa en kultur med aktiverade och engagerade medborgare, vilket står i kontrast till det allt mer passiverande representativa systemet. ---------------------------------------- Väitöskirja tutkii viime vuosisadan poliittisen ja taloudellisen järjestelmän vuorovaikutusta keskittyen erityisesti kolmen viimeisen vuosikymmenen uusliberaaliseen muutokseen. Talouden liberalisoituminen ja ihmisen individualisoituminen edellyttävät myös poliittisen järjestelmän mukautumista. Asteittaisia muutoksia on ollut havaittavissa, kuten kasvava verkosto-organisaatio, toisin sanoen eri alojen kehityspolitiikkaa toteutetaan yhä enemmän eri yhteiskuntasektoreiden vuorovaikutuksena, kun aikaisemmin ainoastaan julkinen sektori vastasi kehityspolitiikan toteuttamisesta. Pohjanmaan maakunnan neljän tapaustutkimuksen avulla osoitetaan, miten vuorovaikutusareena voidaan rakentaa kulloinkin ajankohtaisen asiakokonaisuuden ympärille. Tästä esimerkkinä on Kokkola-Pietarsaari –alueen innovaatioverkosto, johon osallistuvat sekä julkiset että yksityiset sidosryhmät ja joka samalla rikkoo perinteisiä hallinnollisia rajoja. Toinen, kielteisempi esimerkki on Kaskisten alueen rakennemu utosprosessi Metsä-Botnian tehtaan sulkemisen jälkeen. Siellä pienkuntaongelma yhdistettynä siihen, että yksityinen sidosryhmä ei ole ollut velvollinen tekemään yhteistyötä, on johtanut siihen, että ei ole kyetty luomaan alueen pitkäaikaiseen kehittämiseen sitoutunutta vuorovaikutusareenaa. Tämä osoittaa suomalaisen järjestelmän kankeuden, jossa historiallisesta näkökulmasta vahva yhtenäinen valtio ja hajanainen kuntataso estävät joustavan yhteiskuntamuutoksiin sopeutumisen. Väitöskirja osoittaa, että verkosto-organisaation on oltava yhteiskunnassa vaikuttaviin rakenteisiin integroitu osa, jotta se voi vaikuttaa prosessien tuloksiin. Väitöskirja osoittaa ennen kaikkea, että osallistuvilla ja toiminnallisesti määritellyillä alueellisilla ja paikallisilla areenoilla on suuri taloudellinen ja demokraattinen potentiaali. Passivoivan edustavan järjestelmän tilalle on luotava aktiivisten ja tavoitteisiin sitoutuneiden kansalaisten vuorovaikutusjärjestelmä.

« Repertoire general des livres estans en la chambre du procureur du roy nostre sire ou Chastellet de Paris ».

Contient : Les registres dont la table est donnée sont les suivants : « Petit livre blanc » ; « Grand livre blanc » ; « Premier volume » ; « Second volume » ; « Livre vert ancian » ; « Livre noir » ; « Livre vert premier » ; « Livre vert vieil second » ; « Livre rouge vieil » ; « Petit livre jaulne » ; « Petit cayer » ; « Livre verd neuf » ; « Livre bleu » ; « Livre gris » ; « Livre rouge neuf »