Natural killer cell development and function in the context of allogenic stem cell transplantation and cell therapy

Natural killer (NK) cells play a crucial role in the control of viral infections and transformed, cancerous cells. These innate cells have high cytotoxic capacities and use a multitude of receptors to interact with neighbouring cells. The balance of activating and inhibitory signals derived from these receptors determines if NK cells release this cytotoxic potential towards the encountered cell or not. It is this high receptor diversity that defines the phenotype and function of discrete NK cell subsets. This thesis focused on investigating some of the multiple dimensions that influence the function and development of NK cells, with the ultimate aim of improving the current approaches to NK cell immunotherapy against haematological malignancies. NK cell education, the process by which NK cells are effectively trained to recognise the body’s own HLA molecules on healthy cells, is instrumental to generate highly cytotoxic NK cells. When educated NK cells encounter cells missing these self HLA-ligands they lose their inhibition, and the signalling balance is shifted towards activation and killing of the encountered cell. This process is greatly influenced by the interaction of inhibitory KIR receptors and particular HLA molecules. Paper I explores the intimate relationship between allelic variation and NK cell function. It delves into how the polymorphism in KIR and HLA-class I genes influences the loading of granzyme B, a key effector molecule harboured in the secretory lysosomes of NK cells. Employing a high-resolution analysis of these genes' variations across a population of 365 blood donors, we observed that allelic variation in HLA-class I genes is responsible for the observed variation in granzyme B levels, which, in turn, plays a critical role in determining the lytic hit and downstream killing of MHC-deficient target cells. We further showed that NK cell education is stable over time and is independent of age or gender. These findings underscore the significant role genetic factors play in influencing NK cell functionality, laying the groundwork for subsequent research and potential therapeutic developments that could leverage these genetic influences. Haematopoietic stem cell transplantation (HSCT) is a potentially curative treatment for multiple haematological malignancies. A quick and effective reconstitution of the NK cell compartment following transplantation has been shown to correlate with better clinical outcomes and lower relapse incidence. However, immunosuppressive compounds need to be given to the patient to keep the grafted immune system in check and avoid GvHD. Sirolimus, an mTOR inhibitor, has been recognized as an effective component of GvHD prophylaxis regimens. Paper II analyses the effect of sirolimus containing GvHD prophylaxis on the immune reconstitution of transplantation patients. Employing a broad immune cell mapping technique, we uncovered a profound suppression of the naïve CD4 T cell compartment while relatively sparing regulatory T cells and enriching activated CD8 T cells. Given NK cells’ important antileukemic potential, we focused our evaluation on NK cell phenotype, function, and proliferative capacity. The results show that treatment with sirolimus leads to similar immune cell profiles as conventional prophylaxis but leaves a significant impact on NK cell reconstitution. The sirolimus-treated group showed a slight maturation of NK cell subsets and a reduction in their proliferation, and differentiation. This work provides insights into the lasting effects of mTOR inhibition on immune reconstitution and specifically on NK cells following HSCT. And highlight the importance of metabolic regulation in therapeutically modulating NK cell responses during HSCT. The field of NK cell immunotherapy has expanded dramatically in the last decades, however identifying suitable sources of highly potent NK cells has remained a challenge. Allogeneic NK cell therapy, exploiting the missing-self effect of donor educated NK cells to boost their killing potential towards the patients’ leukemic cells, has been shown to have a high potential of inducing remission. Adaptive NK cells have been shown to be a highly educated and potent NK cell subset. In Paper III we developed a GMP-compliant protocol for expanding adaptive NK cells to generate large doses of highly functional and alloreactive ADAPT-NK cells. We demonstrate that ADAPT-NK retain similar transcriptomic and phenotypic features as conventional adaptive NK cells and are potent killers of a wide array of HLA-C/KIR mismatched tumour cell lines and primary AML samples. Finally, combination with NK cell engagers to re-direct their killing broadens their applicability beyond HLA-C/KIR mismatch. The results of this study define a path towards an effective NK cell-based therapy for haematological malignancies and introduces the concept of an ‘’off-the-shelf’’ cell therapy with a non-engineered yet highly specific NK cell population. Collectively, this thesis provides a multi-dimensional exploration of NK cell development and function. It underscores the significant role of genetic variation in modulating NK cell functionality, reveals the lasting impact of mTOR inhibition on NK cell reconstitution following HSCT, and highlights the therapeutic potential of using specially expanded adaptive NK cells for treating haematological malignancies. These insights expand our understanding of NK cell biology in transplantation and cell therapy and could contribute to developing novel therapeutic strategies for treating haematological diseases

Diseño de cockpit para simulador de conducción de competición

En este proyecto se diseña un cockpit de simracing. Se analizará el mercado, se diseñará una estética adecuada, se hará un análisis de la ergonomía para conseguir la mejor posición de conducción y se aplicará una estructura con la suficiente solidez para cumplir los requerimientos del hardware que se usa en esta disciplina. Cockpit: dentro del contexto del simracing se entiende como la estructura que soportara nuestro volante de simulación, pedalera y asiento. Simracing: (Simulated Racing) define la competición en simuladores, podríamos traducirlo como automovilismo virtual, siendo lo mismo que hacen los pilotos reales, pero en el mundo virtual. ¿Qué ventajas tiene? Pues que no tienes que hacer frente a numerosas facturas de ruedas, gasolina, alquiler de circuito y mejor aún, desperfectos y daños personales si tienes algún accidente. Ergonomía: Conjunto de conocimientos de carácter multidisciplinar aplicados para la adecuación de los productos, sistemas y entornos artificialesDepartamento de Ciencias de los Materiales e Ingeniería Metalúrgica, Expresión Gráfica en la Ingeniería, Ingeniería Cartográfica, Geodesia y Fotogrametría, Ingeniería Mecánica e Ingeniería de los Procesos de FabricaciónGrado en Ingeniería en Diseño Industrial y Desarrollo de Product

Analyzing Gerrit Code Review Parameters with Bicho

Code review is becoming a common practice in large scale software development projects. In the case of free, open source software projects, many of them are selecting Gerrit as the system to support the code review process. Therefore, the analysis of the information produced by Gerrit allows for the detailed tracking of the code review process in those projects. In this paper, we present an approach to retrieve and analyze that information based on extending Bicho, a tool designed to retrieve information from issue tracking systems. The details of the retrieval process, the model used to map code review abstractions to issue tracking abstractions, and the structure of the retrieved information are described in detail. In addition, some results of using this approach in a real world scenario, the OpenStack Gerrit code review system, are presented

Habitat differences filter functional diversity of low dispersive microscopic animals (Acari, Halacaridae)

We are starting to appreciate that microscopic animals are not as widespread as previously thought, but we still ignore to what extent and through which mechanisms the environment selects for specific communities or traits in microscopic animals. We here analyse the functional diversity of marine mite communities living in a seagrass meadow across two habitats: the leaves and the matte. The strictly benthic lifestyle and the conserved morphology of mites allow for unambiguous characterisation of their functional traits, while the discrete nature of the two habitats alleviates the uncertainty in their ecological characterisation. Our results show that habitat filters the distribution of certain traits favouring a higher diversity, dispersion, and evenness of functional traits in the matte than in the leaves. We further observed temporal variations in the functional diversity of communities, following the changes in biomass and structure of seagrass leaves. However, despite the stark differences between the two habitats, the filtering effect is partial and affects mostly relative species abundances. Our study emphasises the need of moving from a taxonomical towards a functional view of ecological studies of microscopic organisms. This integrative approach is key to achieve a mechanistic understanding of their habitat and distribution patterns.Peer reviewe

Visualizing hydrophobic and hydrophilic enzyme interactions during immobilization by means of infrared microscopy

A novel Fourier transform infrared (FT-IR) microscopy method was developed and used to analyze the diffusion of lipase CalB in two different resins during immobilization. The method consisted of a streamlined sample preparation process and an automated transmission FT-IR microscopic measurement using a commercial benchtop device. The immobilization of CalB was performed on a hydrophobic resin containing aromatic groups (ECR1030M based on divinylbenzene) and on a hydrophilic resin containing ester groups and thus oxygen (ECR8204M based on methacrylate) and FT-IR revealed that the kinetic of immobilization and the distribution of the enzyme on the two resins were completely different. Furthermore, the technique revealed that CalB was immobilized on the external surface only in the case of the hydrophobic ECR1030M in a layer of about 50–70 μm, whereas when immobilized on the hydrophilic carrier ECR8204M the interaction of the enzyme with the carrier was uniform over the full diameter of the polymer bead. The enzyme activity however was higher on the hydrophobic support ECR1030M

Three-dimensional Reconstruction of the Caspe Geological Structure (Spain) for Evaluation as a Potential CO2 Storage Site

The Caspe geological structure was formed by the convergence of the Iberian Range and the Catalonian Coastal Range, during the Tertiary compression. Traditionally, the Caspe structure has been interpreted from seismic profiles without considering surface structural data. The aim of this study is to build a 3D geological model taking into account the structural data from the geological map, stress fields and lineaments, and evaluate its possibility as potential CO2 storage site. Four surfaces have been modelled: Buntsandstein Top, Muschelkalk-I Top, Muschelkalk-II Top and Cenozoic Bottom. Considering the geometry and depth for storage the target reservoir was considered to be the Buntsandstein facies. The available seismic data indicate that the Buntsandstein facies top is at approximately 500 m depth and hosts a deep saline aquifer. The target reservoir series include the conglomerate and sandstone of the Hoz del Gallo and Cañizar Fms (Buntsandstein Facies) with an average thickness of 500 m and 21% porosity. The seal comprises the shales and silts of the Röt Fm with an average thickness of 100-150 m. The structure volume was calculated based on the -500 mbsl for the Buntsandstein top deepest closed contour lines. The estimated volume is 5, 800 Mm3 with most of CO2 in gaseous state

Parasite Glycobiology:A Bittersweet Symphony

Human infections caused by parasitic protozoans and helminths are among the world's leading causes of death. More than a million people die each year from diseases like malaria and neglected tropical diseases like leishmaniasis, trypanosomiasis, and schistosomiasis. Patients also endure disabilities that cause lifelong suffering and that affect productivity and development [1]. More insidiously, parasites generate important economic losses, since they often also infect commercially valuable animals. Worldwide, exposure to parasites is increasing due to growing international travel and migrations, as well as climate changes, which affect the geographic distribution of the parasite vectors. The parasitic threat is also aggravated by the rise of the immunocompromised population, which is particularly sensitive to parasite infections (e.g., individuals with AIDS and other immunodeficiencies). A common feature of protozoan parasites and helminths is the synthesis of glycoconjugates and glycan-binding proteins for protection and to interact and respond to changes in their environment. To address the many challenges associated with the study of the structure, the biosynthesis, and the biology of parasitic glycans, the authors of this article have established GlycoPar, a European Marie Curie training program steered by some of the world's academic leaders in the field of parasite glycobiology, in close association with European industrial enterprises. The main scientific goal of this network is the description of novel paradigms and models by which parasite glycoconjugates play a role in the successful colonization of the different hosts. By means of a training-through-research program, the aim of the network is to contribute to the training of a generation of young scientists capable of tackling the challenges posed by parasite glycobiology

Preparation of printable and biodegradable cellulose-laponite composite for electronic device application

Printable and biodegradable printed circuit boards (PCBs) prepared by using cellulose as the continuous matrix, laponite as flame retardant filler with various weight ratio (0, 5, 10 and 20 wt% with respect to the α-cellulose quantity used to prepare the composites) and 1-ethyl-3-methylimidazolium acetate ([emim][OAc]) as the recoverable dissolution medium. Prepared cellulose-composites were subjected into physical, chemical, thermal, mechanical and biodegradation analyses to check the suitability of the cellulose-laponite composite for biodegradable electronic application. The addition of laponite into cellulose increased the degradation temperature, flame retardancy and decreased the mechanical properties of the cellulose-laponite composites. The surface nature of the cellulose composite converted from hydrophilic to hydrophobic (contact angle value increased in the range from 50° to 112°) by treating with relatively small amount of hydrophobizing agent (< 1 wt%). The conductive ink printing experiments on the composites explaining the role of hydrophobizing agent and laponite in the composites. Biodegradability of the cellulose was evaluated by enzyme treatments and derived the effect of laponite, hydrophobic agent and conductive ink