Fluctuation-Driven Neural Dynamics Reproduce Drosophila Locomotor Patterns.

The neural mechanisms determining the timing of even simple actions, such as when to walk or rest, are largely mysterious. One intriguing, but untested, hypothesis posits a role for ongoing activity fluctuations in neurons of central action selection circuits that drive animal behavior from moment to moment. To examine how fluctuating activity can contribute to action timing, we paired high-resolution measurements of freely walking Drosophila melanogaster with data-driven neural network modeling and dynamical systems analysis. We generated fluctuation-driven network models whose outputs-locomotor bouts-matched those measured from sensory-deprived Drosophila. From these models, we identified those that could also reproduce a second, unrelated dataset: the complex time-course of odor-evoked walking for genetically diverse Drosophila strains. Dynamical models that best reproduced both Drosophila basal and odor-evoked locomotor patterns exhibited specific characteristics. First, ongoing fluctuations were required. In a stochastic resonance-like manner, these fluctuations allowed neural activity to escape stable equilibria and to exceed a threshold for locomotion. Second, odor-induced shifts of equilibria in these models caused a depression in locomotor frequency following olfactory stimulation. Our models predict that activity fluctuations in action selection circuits cause behavioral output to more closely match sensory drive and may therefore enhance navigation in complex sensory environments. Together these data reveal how simple neural dynamics, when coupled with activity fluctuations, can give rise to complex patterns of animal behavior.

Evolution of reduced post-copulatory molecular interactions in Drosophila populations lacking sperm competition.

In many species with internal fertilization, molecules transferred in the male ejaculate trigger and interact with physiological changes in females. It is controversial to what extent these interactions between the sexes act synergistically to mediate the female switch to a reproductive state or instead reflect sexual antagonism evolved as a by product of sexual selection on males. To address this question, we eliminated sexual selection by enforcing monogamy in populations of Drosophila melanogaster for 65 generations and then measured the expression of male seminal fluid protein genes and genes involved in the female response to mating. In the absence of sperm competition, male and female reproductive interests are perfectly aligned and any antagonism should be reduced by natural selection. Consistent with this idea, males from monogamous populations showed reduced expression of seminal fluid protein genes, 16% less on average than in polygamous males. Further, we identified 428 genes that responded to mating in females. After mating, females with an evolutionary history of monogamy exhibited lower relative expression of genes that were up regulated in response to mating and higher expression of genes that were down-regulated - in other words, their post-mating transcriptome appeared more virgin-like. Surprisingly, these genes showed a similar pattern even before mating, suggesting that monogamous females evolved to be less poised for mating and the accompanying receipt of male seminal fluid proteins. This reduced investment by both monogamous males and females in molecules involved in post-copulatory interactions points to a pervasive role of sexual conflict in shaping these interactions.

Waddlia chondrophila induces systemic infection, organ pathology, and elicits Th1-associated humoral immunity in a murine model of genital infection.

Waddlia chondrophila is a known bovine abortigenic Chlamydia-related bacterium that has been associated with adverse pregnancy outcomes in human. However, there is a lack of knowledge regarding how W. chondrophila infection spreads, its ability to elicit an immune response and induce pathology. A murine model of genital infection was developed to investigate the pathogenicity and immune response associated with a W. chondrophila infection. Genital inoculation of the bacterial agent resulted in a dose-dependent infection that spread to lumbar lymph nodes and successively to spleen and liver. Bacterial-induced pathology peaked on day 14, characterized by leukocyte infiltration (uterine horn, liver, and spleen), necrosis (liver) and extramedullary hematopoiesis (spleen). Immunohistochemistry demonstrated the presence of a large number of W. chondrophila in the spleen on day 14. Robust IgG titers were detected by day 14 and remained high until day 52. IgG isotypes consisted of high IgG2a, moderate IgG3 and no detectable IgG1, indicating a Th1-associated immune response. This study provides the first evidence that W. chondrophila genital infection is capable of inducing a systemic infection that spreads to major organs, induces uterus, spleen, and liver pathology and elicits a Th1-skewed humoral response. This new animal model will help our understanding of the mechanisms related to intracellular bacteria-induced miscarriages, the most frequent complication of pregnancy that affects one in four women.

Long-term persistence of immunity induced by OVA-coupled gas-filled microbubble vaccination partially protects mice against infection by OVA-expressing Listeria.

Vaccination aims at generating memory immune responses able to protect individuals against pathogenic challenges over long periods of time. Subunit vaccine formulations based on safe, but poorly immunogenic, antigenic entities must be combined with adjuvant molecules to make them efficient against infections. We have previously shown that gas-filled microbubbles (MB) are potent antigen-delivery systems. This study compares the ability of various ovalbumin-associated MB (OVA-MB) formulations to induce antigen-specific memory immune responses and evaluates long-term protection toward bacterial infections. When initially testing dendritic cells reactivity to MB constituents, palmitic acid exhibited the highest degree of activation. Subcutaneous immunization of naïve wild-type mice with the OVA-MB formulation comprising the highest palmitic acid content and devoid of PEG2000 was found to trigger the more pronounced Th1-type response, as reflected by robust IFN-γ and IL-2 production. Both T cell and antibody responses persisted for at least 6 months after immunization. At that time, systemic infection with OVA-expressing Listeria monocytgenes was performed. Partial protection of vaccinated mice was demonstrated by reduction of the bacterial load in both the spleen and liver. We conclude that antigen-bound MB exhibit promising properties as a vaccine candidate ensuring prolonged maintenance of protective immunity.

Influence of PTPN2 and pre-existing immunity on the generation of effector and memory CD8 T cells

Notre système immunitaire joue un rôle important pour la protection envers les maladies infectieuses. Au cours d'une réponse à une infection primaire, des cellules B et des cellules T spécifiques, dirigées contre le pathogène en question, sont générées et certaines d'entre elles deviennent des cellules dites mémoires. Leur fonction est de nous protéger contre une nouvelle infection avec le même pathogène, une infection secondaire. Dans certaines situations, comme c'est par exemple le cas avec la grippe, les pathogènes ne sont pas toujours complètement identiques et les cellules mémoires ne sont pas à même d'assurer leur rôle protecteur et d'empêcher une réinfection. Pourtant, on ne sait à l'heure actuelle que très peu comment une immunité acquise, mais non protectrice, influence le développement d'une réponse immunitaire ultérieure. Dans la première partie de cette thèse, nous avons étudié comment les cellules T mémoires cytotoxiques altèrent la réponse de cellules T cytotoxiques nouvellement induites. Au cours d'une réaction immunitaire dirigée contre une infection primaire, un vaste répertoire de lymphocytes T est créé, constitué de cellules T possédant divers degrés d'affinité pour le pathogène. Lors d'une infection secondaire, seules les cellules T ayant une forte affinité pour le pathogène participent à la réponse. Nous avons pu démontrer que ce phénomène de restriction du répertoire des cellules T est principalement causé par les cellules T mémoires qui sont à même de reconnaître un antigène pathogénique présent dans les deux infections. Dans un deuxième projet, nous avons étudié comment l'absence de PTPN2 influence la réponse des cellules T. Chez l'homme, une mutation dans le gène de PTPN2 est associée à des maladies auto-immunes et résulte en une activité réduite de cette phosphatase dans les lymphocytes T. Nous avons montré que la baisse d'activité de la phosphatase PTNP2 conduit à une meilleure expansion des cellules T ayant une qualité comparable à des cellules T auto-antigène spécifiques. De plus, nous avons observé que la survie de ces cellules T effectues ayant une phosphatase diminuée est nettement améliorée. Cela peut conduire à une réponse immunitaire plus efficace ou, éventuellement, à une pathologie auto-immune plus grave. En outre, nos résultats montrent qu'en manipulant l'activité de cette phosphatase, il est possible d'augmenter l'efficacité du transfert des cellules T dans un hôte receveur. Un tel transfert de cellules T est pratiqué chez des patients atteints de tumeurs. Nos travaux suggèrent que la manipulation de la phosphatase PTPN2 pourrait donc représenter une approche thérapeutique novatrice et prometteuse. -- Notre système immunitaire joue un rôle important pour la protection contre les maladies. Les cellules T CD8+ ont une importance primordiale pour le contrôle d'infections primaires causées par des virus ou bactéries, mais également contre certaines tumeurs. Par conséquent, mieux comprendre les exigences nécessaires à l'induction de bonnes réponses des cellules T CD8 pourrait nous permettre de construire des vaccins contre les pathogènes contre lesquels nous n'avons pour l'instant pas de vaccins mais aussi d'améliorer les réactions immunitaires dirigées anti-tumorales. Dans la première partie de cette thèse, nous avons étudié l'influence qu'une immunité préexistante a sur la réponse des cellules T CD8. Nous sommes souvent exposés à des pathogènes qui sont similaires mais pas identiques à ceux que nous avons rencontrés auparavant. De telles infections hétérologues ne sont pas l'objet de beaucoup d'études et certains exemples indiquent même qu'une immunité préexistante partielle peut mener à une aggravation de la maladie. Nous avons étudié le répertoire des lymphocytes T CD8 qui sont générés lors d'une rencontre avec un nouvel antigène, et ce en comparant infection primaire et secondaire. En utilisant le modèle expérimental d'infections à Listeria monocytogenes, nous avons pu montrer que lors d'une infection primaire, un répertoire diversifié comprenant des cellules T CD8 de forte et faible affinité est constitué. Au contraire, dans le cas d'une infection secondaire, le répertoire des cellules T est fortement limité et seulement les lymphocytes T de forte affinité sont impliqués dans la réponse immunitaire. Nous avons pu démontrer que ces Rangements sont provoqués par des cellules T CD8 mémoires capables de reconnaître un antigène présent dans les deux infections. Cette augmentation du seuil d'activation des cellules effectrices est majoritairement causée par les lymphocytes T CD8 mémoires non transférables. Ces observations indiquent que les vaccins visant à induire des cellules T anti-tumorales de faible affinité seraient inefficaces si le vaccin contient des épitopes contre lesquels il existe une mémoire immunologique. Les réponses immunitaires conduites par les cellules T contre les antigènes tumoraux dépendent des cellules T CD8 de faible réactivité contre les antigènes tumoraux puisque les cellules à forte réactivité sont éliminées par les mécanismes de tolérance. Nous basant sur l'existence dans la littérature de preuves indiquant que PTPN2 influence la réponse des cellules T de faible affinité, nous nous sommes intéressés à comprendre comment PTPN2 impacte les réponses des cellules T CD8 en général. Nous avons remarqué que des cellules T CD8 déficientes en PTPN2 exhibent une meilleure capacité à proliférer suite à une faible ou courte stimulation du récepteur des lymphocytes T. La phase effectrice est prolongée et la contraction retardée résultant ainsi à globalement plus de cellules effectrices. Ce phénomène est également accompagné d'une meilleure survie des cellules effectrices de différentiation terminale. Une fois transférées dans un nouvel hôte receveur, les cellules effectrices terminales KLRG1+CD127- déficientes en phosphatase PTPN2 peuvent survivre et se transformer en cellules mémoires CD127+ fonctionnelles. De façon inattendue, nous avons découvert que l'élimination de PTPN2 améliore l'efficacité du transfert et la formation des cellules mémoires ainsi que leur capacité protectrice. Manipuler l'activité de cette phosphatase apparaît donc comme une approche intéressante et prometteuse pour la thérapie cellulaire par transfert adoptif de lymphocytes T. Nos observations montrent que la manipulation d'un facteur intrinsèque, l'absence de PTPN2, peut, dans certaines circonstances, améliorer la réponse des cellules T. Une meilleure connaissance des mécanismes contrôlant la réponse des lymphocytes T CD8 pourrait donc permettre la manipulation de ces derniers et conduire à des réponses immunitaires plus vigoureuses. Si ces réponses sont déclenchées par l'utilisation de vaccins, il est nécessaire de considérer l'historique d'une exposition préalable à des agents pathogènes ou à des vaccins puisque celle-ci peut, comme nous l'avons démontré, influencer le répertoire des cellules T recrutées dans la réponse immunitaire et, par conséquent, modifier l'aptitude de notre système immunitaire à faire face à une infection. -- Our immune system plays an important role in the protection from disease. CD8 T cells are critical for the control of primary infections with most viruses and certain bacteria as well as against some tumors. Therefore, better knowledge of CD8 T cell responses might enable us to generate vaccines against pathogens for which currently no vaccines are available or to improve anti-tumor immune responses. In the first part of this thesis we addressed the issue how previously acquired immunity impacts on the response of CD8 T cells. We are often exposed to pathogens that are related but not identical to the previously encountered ones. Such heterologous infections are not well studied and there are some indications that partial pre-existing immunity may in some cases even lead to an enhancement of disease. We specifically studied the T cell repertoire of CD8 T cells that are responding to a newly encountered antigen in secondary compared to primary infections. Using the experimental model of Listeria monocytogenes infections, we showed that in primary infections a wide repertoire including high and low affinity CD8 T cells is recruited into the immune response. In contrast to this, in secondary infections, the T cell repertoire is severely restricted and only T cells of high affinity are responding. We were able to pinpoint this difference to the presence of memory CD8 T cells that recognize an antigen that is shared between the two subsequent infections. This increase in the activation threshold was most effectively mediated via non-transferable memory CD8 T cells. This would argue that vaccines targeting low affinity tumor-specific T cells would fail if the vaccine contains previously encountered CD8 T cell epitopes. T cell mediated immune responses to tumor antigen rely often on T cells which weakly react to tumor antigen as high affinity T cells are eliminated by tolerance mechanisms. Following indication in the literature that PTPN2 impacts on the response of such weakly antigen-reactive T cells, we investigated how PTPN2 impacts in general the response of CD8 T cells. We observed that CD8 T cells lacking PTPN2 show an enhanced expansion following weak or short-term T cell receptor stimulation. The effector phase is prolonged and contraction delayed thus resulting in overall more effector cells. This is accompanied by a better survival of terminal effector cells. When transferred into new recipients, KLRG1+CD127- terminal effector cells lacking PTPN2 can survive and convert into CD127+ functional memory cells. Surprisingly, we discovered that elimination of PTPN2 enhances the transfer efficacy and formation of memory cells as well as the protective capacity. Targeting PTPN2 might thus be a promising approach for adoptive T cell therapy. Our observations show how the manipulation of an intrinsic factor, the absence of PTPN2, can enhance T cell responses under certain circumstances. A better understanding of underlying mechanisms for the control of CDS T cell responses might enable the manipulation of these and allow for more powerful responses. If these responses are induced through vaccines it is imperative that the previous history of exposure to pathogens or vaccines is considered as it can, as we have shown in this thesis, influence the recruited T cell repertoire and thus possibly the ability to handle the infection.

Identificació de nous gens que participen en la remodelació del sistema traqueal de Drosophila melanogaster

Un dels organismes model més utilitzats en experimentació genètica és la Drosophila melanogaster ja que la facilitat de manipulació genètica i la seva simplicitat permeten estudiar processos biològics amb múltiples aplicabilitats en diferents àmbits d’estudi com el desenvolupament embrionari i la morfogènesis. La morfogènesi es un dels esdeveniments més importants durant el desenvolupament embrionari que permet la formació dels diferent teixits i òrgans, i que depèn de l'expressió genètica i de l'activació i coordinació de diferents vies de senyalització. Entendre com es coordinen aquest processos es fonamental per conèixer com es forma un òrgan. Així, l’objectiu principal d’aquest Treball de Final de Grau és identificar nous gens implicats en la formació del sistema traqueal (el nostre òrgan model) mitjançant un mini-­‐cribratge funcional de gens que s’expressen en la tràquea, a més de generar eines per a l'estudi de la via de senyalització FGF/Bnl durant la remodelació del sistema traqueal mitjançant la tècnica de knock in. Per a dur-­‐ho a terme, amb el suport de la base de dades de Gens i Genomes de Drosophila melanogaster (mod-­‐ENCODE Tissue Expression Data) s’han seleccionat gens candidats expressats a la tràquea en estat larvari. Un cop identificats, s'ha estudiat la seva possible funció en el desenvolupament de les tràquees mitjançant el seu silenciament amb el sistema UAS-­‐Gal4. Així hem vist que Vein (CG10491), CG17098, No Ocelli (CG4491) i Peptidasa (CG4017) presenten diversos fenotips que afecten la formació dels traqueoblasts. També hem vist que Vein, lligand de la via EGF és necessari per a la proliferació i supervivència de les cèl·∙lules traqueals del sac aeri. Finalment s’ha iniciat la generació d'un knock in en el gen branchless (bnl). Per aquest motiu s'han amplificat les regions 5’ i 3’ de l’exó 2 del gen Bnl i s'ha iniciat la seva clonació dirigida al vector de destí pTV-­‐Cherry. Aquesta tècnica generarà eines que permetran entendre la funció del gen bnl durant la remodelació del sistema traqueal.

Delivery of antigen to nasal-associated lymphoid tissue microfold cells through secretory IgA targeting local dendritic cells confers protective immunity.

BACKGROUND: Transmission of mucosal pathogens relies on their ability to bind to the surfaces of epithelial cells, to cross this thin barrier, and to gain access to target cells and tissues, leading to systemic infection. This implies that pathogen-specific immunity at mucosal sites is critical for the control of infectious agents using these routes to enter the body. Although mucosal delivery would ensure the best onset of protective immunity, most of the candidate vaccines are administered through the parenteral route. OBJECTIVE: The present study evaluates the feasibility of delivering the chemically bound p24gag (referred to as p24 in the text) HIV antigen through secretory IgA (SIgA) in nasal mucosae in mice. RESULTS: We show that SIgA interacts specifically with mucosal microfold cells present in the nasal-associated lymphoid tissue. p24-SIgA complexes are quickly taken up in the nasal cavity and selectively engulfed by mucosal dendritic cell-specific intercellular adhesion molecule 3-grabbing nonintegrin-positive dendritic cells. Nasal immunization with p24-SIgA elicits both a strong humoral and cellular immune response against p24 at the systemic and mucosal levels. This ensures effective protection against intranasal challenge with recombinant vaccinia virus encoding p24. CONCLUSION: This study represents the first example that underscores the remarkable potential of SIgA to serve as a carrier for a protein antigen in a mucosal vaccine approach targeting the nasal environment.

Multimodal stimulus coding by a gustatory sensory neuron in Drosophila larvae.

Accurate perception of taste information is crucial for animal survival. In adult Drosophila, gustatory receptor neurons (GRNs) perceive chemical stimuli of one specific gustatory modality associated with a stereotyped behavioural response, such as aversion or attraction. We show that GRNs of Drosophila larvae employ a surprisingly different mode of gustatory information coding. Using a novel method for calcium imaging in the larval gustatory system, we identify a multimodal GRN that responds to chemicals of different taste modalities with opposing valence, such as sweet sucrose and bitter denatonium, reliant on different sensory receptors. This multimodal neuron is essential for bitter compound avoidance, and its artificial activation is sufficient to mediate aversion. However, the neuron is also essential for the integration of taste blends. Our findings support a model for taste coding in larvae, in which distinct receptor proteins mediate different responses within the same, multimodal GRN.

Genomics of clinal variation in Drosophila: disentangling the interactions of selection and demography.

Clines in phenotypes and genotype frequencies across environmental gradients are commonly taken as evidence for spatially varying selection. Classical examples include the latitudinal clines in various species of Drosophila, which often occur in parallel fashion on multiple continents. Today, genomewide analysis of such clinal systems provides a fantastic opportunity for unravelling the genetics of adaptation, yet major challenges remain. A well-known but often neglected problem is that demographic processes can also generate clinality, independent of or coincident with selection. A closely related issue is how to identify true genic targets of clinal selection. In this issue of Molecular Ecology, three studies illustrate these challenges and how they might be met. Bergland et al. report evidence suggesting that the well-known parallel latitudinal clines in North American and Australian D. melanogaster are confounded by admixture from Africa and Europe, highlighting the importance of distinguishing demographic from adaptive clines. In a companion study, Machado et al. provide the first genomic comparison of latitudinal differentiation in D. melanogaster and its sister species D. simulans. While D. simulans is less clinal than D. melanogaster, a significant fraction of clinal genes is shared between both species, suggesting the existence of convergent adaptation to clinaly varying selection pressures. Finally, by drawing on several independent sources of evidence, Bo?ičević et al. identify a functional network of eight clinal genes that are likely involved in cold adaptation. Together, these studies remind us that clinality does not necessarily imply selection and that separating adaptive signal from demographic noise requires great effort and care.

Bartonella apis sp. nov., a honey bee gut symbiont of the class Alphaproteobacteria.

Here, we report the culture and characterization of an alphaproteobacterium of the order Rhizobiales, isolated from the gut of the honey bee Apis mellifera. Strain PEB0122T shares >95 % 16S rRNA gene sequence similarity with species of the genus Bartonella, a group of mammalian pathogens transmitted by bloodsucking arthropods. Phylogenetic analyses showed that PEB0122T and related strains from the honey bee gut form a sister clade of the genus Bartonella. Optimal growth of strain PEB0122T was obtained on solid media supplemented with defibrinated sheep blood under microaerophilic conditions at 35-37 °C, which is consistent with the cultural characteristics of other species of the genus Bartonella. Reduced growth of strain PEB0122T also occurred under aerobic conditions. The rod-shaped cells of strain PEB0122T had a mean length of 1.2-1.8 μm and revealed hairy surface structures. Strain PEB0122T was positive for catalase, cytochrome c oxidase, urease and nitrate reductase. The fatty acid composition was comparable to those of other species of the genus Bartonella, with palmitic acid (C16 : 0) and isomers of 18- and 19-carbon chains being the most abundant. The genomic DNA G+C content of PEB0122T was determined to be about 45.5 mol%. The high 16S rRNA gene sequence similarity with species of Bartonella and its close phylogenetic position suggest that strain PEB0122T represents a novel species within the genus Bartonella, for which we propose the name Bartonella apis sp. nov. The type strain is PEB0122T ( = NCIMB 14961T = DSM 29779T).

Synthetic long peptide-based vaccine formulations for induction of cell mediated immunity: A comparative study of cationic liposomes and PLGA nanoparticles.

Nanoparticulate formulations for synthetic long peptide (SLP)-cancer vaccines as alternative to clinically used Montanide ISA 51- and squalene-based emulsions are investigated in this study. SLPs were loaded into TLR ligand-adjuvanted cationic liposomes and PLGA nanoparticles (NPs) to potentially induce cell-mediated immune responses. The liposomal and PLGA NP formulations were successfully loaded with up to four different compounds and were able to enhance antigen uptake by dendritic cells (DCs) and subsequent activation of T cells in vitro. Subcutaneous vaccination of mice with the different formulations showed that the SLP-loaded cationic liposomes were the most efficient for the induction of functional antigen-T cells in vivo, followed by PLGA NPs which were as potent as or even more than the Montanide and squalene emulsions. Moreover, after transfer of antigen-specific target cells in immunized mice, liposomes induced the highest in vivo killing capacity. These findings, considering also the inadequate safety profile of the currently clinically used adjuvant Montanide ISA-51, make these two particulate, biodegradable delivery systems promising candidates as delivery platforms for SLP-based immunotherapy of cancer.

Postmating-prezygotic isolation between two allopatric populations of Drosophila montana: fertilisation success differs under sperm competition

Postmating but prezygotic (PMPZ) interactions are increasingly recognized as a potentially important early-stage barrier in the evolution of reproductive isolation. A recent study described a potential example between populations of the same species: single matings between Drosophila montana populations resulted in differential fertilisation success because of the inability of sperm from one population (Vancouver) to penetrate the eggs of the other population (Colorado). As the natural mating system of D. montana is polyandrous (females remate rapidly), we set up double matings of all possible crosses between the same populations to test whether competitive effects between ejaculates influence this PMPZ isolation. We measured premating isolation in no-choice tests, female fecundity, fertility and egg-to-adult viability after single and double matings as well as second-male paternity success (P-2). Surprisingly, we found no PMPZ reproductive isolation between the two populations under a competitive setting, indicating no difficulty of sperm from Vancouver males to fertilize Colorado eggs after double matings. While there were subtle differences in how P-2 changed over time, suggesting that Vancouver males' sperm are somewhat less competitive in a first-male role within Colorado females, these effects did not translate into differences in overall P-2. Fertilisation success can thus differ dramatically between competitive and noncompetitive conditions, perhaps because the males that mate second produce higher quality ejaculates in response to sperm competition. We suggest that unlike in more divergent species comparisons, where sperm competition typically increases reproductive isolation, ejaculate tailoring can reduce the potential for PMPZ isolation when recently diverged populations interbreed.

Mode of interaction of the Gαo subunit of heterotrimeric G proteins with the GoLoco1 motif of Drosophila Pins is determined by guanine nucleotides.

Drosophila GoLoco motif-containing protein Pins is unusual in its highly efficient interaction with both GDP- and the GTP-loaded forms of the α-subunit of the heterotrimeric Go protein. We analysed the interactions of Gαo in its two nucleotide forms with GoLoco1-the first of the three GoLoco domains of Pins-and the possible structures of the resulting complexes, through combination of conventional fluorescence and FRET measurements as well as through molecular modelling. Our data suggest that the orientation of the GoLoco1 motif on Gαo significantly differs between the two nucleotide states of the latter. In other words, a rotation of the GoLoco1 peptide in respect with Gαo must accompany the nucleotide exchange in Gαo. The sterical hindrance requiring such a rotation probably contributes to the guanine nucleotide exchange inhibitor activity of GoLoco1 and Pins as a whole. Our data have important implications for the mechanisms of Pins regulation in the process of asymmetric cell divisions.

From T cell “exhaustion” to anti-cancer immunity

The immune system has the potential to protect from malignant diseases for extended periods of time. Unfortunately, spontaneous immune responses are often inefficient. Significant effort is required to develop reliable, broadly applicable immunotherapies for cancer patients. A major innovation was transplantation with hematopoietic stem cells from genetically distinct donors for patients with hematologic malignancies. In this setting, donor T cells induce long-term remission by keeping cancer cells in check through powerful allogeneic graft-versus-leukemia effects. More recently, a long awaited breakthrough for patients with solid tissue cancers was achieved, by means of therapeutic blockade of T cell inhibitory receptors. In untreated cancer patients, T cells are dysfunctional and remain in a state of T cell "exhaustion". Nonetheless, they often retain a high potential for successful defense against cancer, indicating that many T cells are not entirely and irreversibly exhausted but can be mobilized to become highly functional. Novel antibody therapies that block inhibitory receptors can lead to strong activation of anti-tumor T cells, mediating clinically significant anti-cancer immunity for many years. Here we review these new treatments and the current knowledge on tumor antigen-specific T cells.

The Drosophila TNF Eiger Is an Adipokine that Acts on Insulin-Producing Cells to Mediate Nutrient Response.

Adaptation of organisms to ever-changing nutritional environments relies on sensor tissues and systemic signals. Identification of these signals would help understand the physiological crosstalk between organs contributing to growth and metabolic homeostasis. Here we show that Eiger, the Drosophila TNF-α, is a metabolic hormone that mediates nutrient response by remotely acting on insulin-producing cells (IPCs). In the condition of nutrient shortage, a metalloprotease of the TNF-α converting enzyme (TACE) family is active in fat body (adipose-like) cells, allowing the cleavage and release of adipose Eiger in the hemolymph. In the brain IPCs, Eiger activates its receptor Grindelwald, leading to JNK-dependent inhibition of insulin production. Therefore, we have identified a humoral connexion between the fat body and the brain insulin-producing cells relying on TNF-α that mediates adaptive response to nutrient deprivation.