| 1. |
- Henningsson, Anna J, 1972-
(author)
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Clinical, epidemiological and immunological aspects of Lyme borreliosis with special focus on the role of the complement system
- 2011
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Doctoral thesis (other academic/artistic)abstract
- Lyme borreliosis (LB) is the most common vector-borne disease in the Northern Hemisphere. The infection is caused by spirochetes belonging to the Borrelia burgdorferi sensu lato complex, and it is transmitted to humans by ticks. LB is associated with several clinical manifestations, of which erythema migrans (EM) and neuroborreliosis (NB) are the most common inEurope. The course of the disease is usually benign, but can vary between individuals. The underlying pathogenic mechanisms are not fully understood, but the prognosis is probably determined by a complex interplay between the bacteria and the host’s immune response. Previous studies have indicated that a strong initial T helper (Th) 1-response followed by a Th2 response is beneficial for the clinical outcome in LB.The aims of this thesis were to follow the incidence of NB inJönköping County,Sweden, over time, to search for clinical and laboratory markers associated with the risk of developing long-lasting post-treatment symptoms, and to explore the role of the complement system as well as the relative balance between Th-associated cytokine/chemokine responses in LB.The number of NB cases, diagnosed by cerebrospinal fluid (CSF) analysis, increased from 5 to 10/100,000 inhabitants/year in Jönköping County during 2000-2005. Post-treatment symptoms persisting more than 6 months occurred in 13 %, and were associated with higher age, longer-lasting symptoms prior to treatment, higher levels of Borrelia-specific IgG in CSF, and reported symptoms of radiculitis. Facial palsy, headache and fever were frequent manifestations in children, whereas unspecific muscle and joint pain were the most commonly reported symptoms in older patients.Complement activation occurred both locally in the skin in EM and in CSF of NB patients. However, no activation could be detected in blood in NB patients. Elevated levels of C1q, C4 and C3a in CSF, along with correlation between C1q and C3a levels, suggest complement activation via the classical pathway locally in the central nervous system in NB. In vitro experiments with two clinical Borrelia isolates revealed that B. garinii LU59 induced higher complement activation in human plasma compared to B. afzelii K78 that recruited more of complement regulator factor H. To elucidate the role of complement in the phagocytosis process, experiments were performed using whole blood from healthy donors incubated with fluorescence-labelled spirochetes and different complement inhibitors. The results illustrated a central role of complement for phagocytosis of Borrelia spirochetes.We also studied the relative contribution of different Th-associated cytokines/chemokine responses in NB. The results support the notion that early NB is dominated by a Th1 response, eventually accompanied by a Th2 response. IL-17A was increased in CSF in half of the patients with confirmed NB, suggesting a hitherto unknown role of Th17 in NB.In conclusion, the risk of developing long-lasting post-treatment symptoms tend to increase mainly with age and duration of symptoms prior to treatment in NB. The complement system seems to play an important role in host defence to recognize and kill Borrelia spirochetes. However, complement activation in inappropriate sites or to an excessive degree may cause tissue damage, and therefore, the role of complement in relation to disease course needs to be studied further. Likewise, the role of Th17 in LB pathogenesis and host defence should be further evaluated in prospective studies.
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| 2. |
- Nilsson, Per H., 1980-
(author)
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Interactions between platelets and complement with implications for the regulation at surfaces
- 2012
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Doctoral thesis (other academic/artistic)abstract
- Disturbances of host integrity have the potential to evoke activation of innate immunologic and hemostatic protection mechanisms in blood. Irrespective of whether the activating stimulus is typically immunogenic or thrombotic, it will generally affect both the complement system and platelets to a certain degree. The theme of this thesis is complement and platelet activity, which is intersected in all five included papers. The initial aim was to study the responses and mechanisms of the complement cascade in relation to platelet activation. The secondary aim was to use an applied approach to regulate platelets and complement on model biomaterial and cell surfaces. Complement activation was found in the fluid phase in response to platelet activation in whole blood. The mechanism was traced to platelet release of stored chondroitin sulfate-A (CS-A) and classical pathway activation via C1q. C3 was detected at the platelet surface, though its binding was independent of complement activation. The inhibitors factor H and C4-binding protein (C4BP) were detected on activated platelets, and their binding was partly dependent on surface-exposed CS-A. Collectively, these results showed that platelet activation induces inflammatory complement activation in the fluid phase. CS-A was shown to be a central molecule in the complement-modulatory functions of platelets by its interaction with C1q, C4BP, and factor H.Platelet activation and surface adherence were successfully attenuated by conjugating an ADP-degrading apyrase on a model biomaterial. Only minor complement regulation was seen, and was therefore targeted specifically on surfaces and cells by co-immobilizing a factor H-binding peptide together with the apyrase. This combined approach led to a synchronized inhibition of both platelet and complement activation at the interface of biomaterials/xenogeneic cells and blood.In conclusion, here presents a novel crosstalk-mechanism for activation of complement when triggering platelets, which highlights the importance of regulating both complement and platelets to lower inflammatory events. In addition, a strategy to enhance the biocompatibility of biomaterials and cells by simultaneously targeting ADP-dependent platelet activation and the alternative complement C3-convertase is proposed.
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| 3. |
- Carlsson, Hanna, 1978-
(author)
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Laboratory methods for investigation of the immunological orchestra in response to pathogens
- 2023
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Doctoral thesis (other academic/artistic)abstract
- Laboratory methods used for investigation of immune response often involve collection of whole blood and analysis of different biomarkers in blood components or generated from pathogen stimulation of whole blood or peripheral blood mononuclear cells (PBMC). Methods used to measure biomarkers are for example enzyme-linked immunosorbent assay (ELISA) which measures one biomarker at a time or multiplex assays for example x-unknown, multi-analyte profiling (xMAP) by Luminex or proximity extension assay (PEA), which can measure up to just over 3000 biomarkers at a time. Analysis of one biomarker at a time are time consuming, costly, and dependent of a large sample size to enable repeated measurements of different analytes. Therefore, multiplex assays that are time saving, more cost effective and measures multiple bi-omarkers at once in a small sample can be applied. The aim of this thesis was to evaluate multiplex laboratory methods for investigation of the immunological orchestra in response to Borrelia infection and influenza immunisation and if possible, further characterize individuals with different clinical outcomes or serological response, respectively. In our studies (paper I-III) we included 1113 blood donors of which 66 were found to previously have had a subclinical borreliosis (defined as presence of Borrelia-specific antibodies without recall of previous Lyme borreliosis), of the 66 individuals 60 were available for participation. We also included 22 patients previously diagnosed with Lyme neuroborreliosis (LNB). In paper IV we included in total 73 individuals consisting of healthcare workers and patients attending seasonal influenza vaccination. We applied whole blood, PBMC and plasma stimulations and measured a range of cytokines, chemokines and complement factors with ELISA, nephelometry, xMAP and PEA. Our results show that subclinical Lyme borreliosis (SB) individuals display the following pattern, low age, male sex, low amount of secreted interleukin (IL)-17, CCL20 and higher secretion of IL-10 by PBMCs stimulated three days with Borrelia garinii compared to patients with previous Lyme neuroborreliosis (LNB). The subclinical individuals also show higher activation of the complement system in response to Borrelia afzelii. We performed multiplex analysis of complement factors in attempt to further characterize our SB individuals and LNB patient but found the results to deviate largely from reference values retrieved with other standardized methods. This highlights the importance of critical review of generated results from all form of assays. To investigate immune responses after influenza immunisation and further characterize serological responders and nonresponders we included measurement of influenza-specific antibodies and total immunoglobulins (Ig) in blood serum, influenza-specific mucosal IgA (nasal-swabs) and cell-mediated immune response in supernatants from PBMCs stimulated with influenza vaccine using PEA. We found the serological responders to be characterised by lower levels of total IgM, Granzyme B (GZMB) and IL-12 together with higher levels of CXCL13 compared with nonresponders. To conclude, xMAP and PEA are two valuable methods that can be applied together with multivariate statistical methods in the investigation of both innate and adaptive immunity characteristics and association to clinical outcome or serological response after Borrelia infection and influenza immunisation, respectively.
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| 4. |
- Hamad, Osama A., 1978-
(author)
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Crosstalk Between Activated Platelets and the Complement System
- 2010
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Doctoral thesis (other academic/artistic)abstract
- Several studies have shown that complement and thrombotic events co-exist. Platelets have been suspected to act as the bridge between the two cascade systems. To study the platelet-induced complement activation we developed a system in which platelets were activated by thrombin receptor activating peptide (TRAP) in platelet rich plasma (PRP) or whole blood anti-coagulated using the specific thrombin inhibitor, lepirudin. TRAP-activated platelets induced a fluid-phase complement activation measured as generation of C3a and sC5b-9, triggered by released chondroitin sulphate-A (CS-A) which interacted with C1q and activated the complement system through the classical pathway. Complement components C1q, C3, C4 and C9 were also shown to bind to TRAP-activated platelets but this binding did not seem to be due to a complement activation since blocking of complement activation at the C1q or C3 levels did not affect the binding of the complement proteins. The C3 which bound to activated platelets consisted of C3(H2O), indicating that bound C3 was not proteolytically activated. Binding of C1q was partially dependent on CS-A exposure on activated platelets. The abolished complement activation on the surface of activated platelets was suggested to be dependent on the involvement of several complement inhibitors. We confirmed the binding of C1INH and factor H to activated platelets. To this list we have added another potent complement inhibitor, C4BP. The binding of factor H and C4BP was shown to be dependent on exposure of CS-A on activated platelets. The physiological relevance of these reactions was reflected in an elevated expression of CD11b on leukocytes, and increased generation of platelet-leukocyte complexes. The platelets were involved in these events by at least two different mechanisms; generation of C5a which activated leukocytes and binding of C3(H2O)/iC3(H2O), a ligand to the intergrin CD11b/CD18 on their surface. These mechanisms add further to the understanding of how platelets interact with the complement system and will help us to understand the role of the complement system in cardiovascular disease and thrombotic conditions.
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| 5. |
- Lindau, Robert, 1989-
(author)
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Immune regulation at the foetal-maternal interface; implications for healthy and complicated pregnancies
- 2020
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Doctoral thesis (other academic/artistic)abstract
- For a successful pregnancy, the maternal immune system must acquire tolerance towards the paternal antigens present in the semi-allogeneic foetus. This tolerance is mainly established locally at the foetal-maternal interface, where foetally-derived trophoblasts invade the maternal endometrium (called decidua during pregnancy) and come in close proximity to maternal immune cells. The decidua is populated by maternal immune cells of a unique composition, characterised by their suppressive phenotypes that are essential for maintaining tissue homeostasis. Accordingly, failure of immune tolerance can lead to pregnancy complications. Macrophages and regulatory T-cells are enriched in the decidua and are believed to play important roles in the establishment of tolerance. However, there is limited information regarding the factors that regulate their functions and if their function is compromised in pregnancy complications.The aim of this thesis was to further elucidate which factors are responsible for induction of the regulatory phenotypes of macrophages and T-cells found in the decidua, how tissue resident cells in the decidua contribute to this and if this system is compromised during pregnancy complications, such as preeclampsia and recurrent pregnancy loss.Decidual stromal cells (DSCs) constitute the largest population of tissue resident cells in the decidua. In an in vitro system of macrophage differentiation, we found that Isolated peripheral blood monocytes cultured in conditioned medium (CM) from DSCs acquired a high expression of the regulatory M2 markers CD163, CD209 and CD14, and a low expression of CD86, characteristics of decidual macrophages. This induction was in part mediated by macrophage-colony stimulating factor (M-CSF), as neutralising its effects reduced the expression of CD163. However, since only a partial reduction was reached, other factors are involved. Another likely candidate for this polarisation is interleukin (IL)-34, a second ligand for the M-CSF receptor. We showed that IL-34 is expressed by both DSCs and the foetal placenta. Further, in vitro, IL-34 was able to induce macrophages with similar properties as that of M-CSF-induced macrophages, with high expression of CD163, CD209 and CD14. This was also coupled to a cytokine secretion profile similar to M-CSF-induced macrophages, with high production of IL-10, low production of tumour necrosis factor (TNF) and no production of IL-12. We found no evidence of IL-34 being aberrantly expressed in placentas from preeclamptic women.In addition to promoting induction of macrophages with a regulatory phenotype, CM from DSCs promoted expansion of Foxp3+CD25bright regulatory T (Treg) cells in an in vitro polarisation system, in a SMAD3 dependent manner. Protein profiling of DSCs revealed limited production of the Th2 related IL-13, IL-4, IL-33 and thymic stromal lymphopoietin (TSLP), as well as no production of the Th17 related IL-17A and chemokine (C-C motif) ligand (CCL) 20. Instead we found that DSCs were more prone to production of regulatory factors, such as M-CSF, leukaemia inhibitory factor (LIF) and transforming growth factor (TGF)-β, albeit with addition of the more pro-inflammatory IL-6, chemokine (C-X-C motif) ligand (CXCL) 8 and the Th1-related CXCL10.We also investigated if the placenta´s ability to induce Treg cells and regulatory M2 macrophages is compromised in women with a history of unexplained recurrent pregnancy loss (uRPL) and if the placental secreted protein profile is skewed to a pro-inflammatory response in uRPL. Using surplus materials from chorionic villous sampling (CVS), we generated CM from placental tissue taken from healthy and uRPL pregnancies and used this to polarise macrophages and T-cells in vitro. We found no difference in the ability to induce Treg cells and regulatory M2 macrophages between the healthy group and the uRPL group. Likewise, no differences in the protein profile was observed between the two groups.Taken together, our findings imply that DSCs produce a variety of factors promoting foetal tolerance by induction of Treg cells and regulatory M2 macrophages. Furthermore, we also showed that the placenta retained its ability to induce Treg cells and regulatory M2 macrophages in women with a history of uRPL.
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| 6. |
- Chapman, David, 1972-
(author)
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Urban design of winter cities : Winter season connectivity for soft mobility
- 2018
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Doctoral thesis (other academic/artistic)abstract
- All across the world the form of the built environment is playing a crucial role as enabler or inhibitor for urban outdoor activity such as soft mobility. Urban form can make it more attractive for people to be mobile outdoors and playing a role in the public life, or it can put people off venturing outside. For winter cities, a question for urban design is how we can design environments that are attractive for outdoor activity in the winter season as well as summer and additionally how will climate change influence these aspects.The reason for studying this is the importance of understanding how, in relation to urban form, weather, seasonal variations, and climate change influences human outdoor activity. In this study the focus on outdoor activity is problematised around the concern that people spend a low percentage of their time outdoors in winter conditions. For society, the problem is that this trend and the related low levels of physical activity are associated with a range of health issues.To study this the main question for this research is what attracts and hinders soft mobility during the winter season and how can this knowledge underpin new considerations about urban design for connectivity in winter cities? To address this, the research methods focused on document studies, surveys, mental mapping, photo elicitation and semi-structured discussions.The study works at three scientific levels. Firstly, it seeks to understand the interrelationship between the built environment and people’s outdoor activity in winter. Secondly, it attempts to understand how connectivity for soft mobility in winter is being affected by weather and climate change. Thirdly, it seeks new ways of thinking about how the urban form can be designed to increase outdoor soft mobility in winter.The discussion and conclusions focused on the argument that in winter settlements, the winter season can alter spatial patterns and settlement organisation. Here it was argued that in these settlements the winter season can be an aspect of urban morphology and can be part of the process of shaping the public realm and its connectivity for soft mobility in winter.
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| 7. |
- Nilsson, Kristina, 1976-
(author)
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Hybrid Methods for Coreference Resolution in Swedish
- 2010
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Doctoral thesis (other academic/artistic)abstract
- The aim of this thesis is to improve coreference resolution in Swedish by providing a hybrid approach based on combining data-driven methods and linguistic knowledge. Coreference resolution here consists in identifying all expressions in a text that have the same referent, for example, a person or an object. The linguistic knowledge is based on Accessibility Theory (Ariel 1990). This is used for guiding the selection of likely anaphor-antecedent pairs from the set of all possible such pairs in a text. The data-driven method adopted is Memory-Based Learning (MBL), a supervised method based on the idea that learning means storing experiences in memory, and that new problems are solved by reusing solutions from similar experiences (Daelemans and Van den Bosch 2005). The referring expressions covered by the system are names, definite descriptions, and pronouns. In order to maximize performance, we use different classifiers with a specific set of linguistically motivated features for each type of expression. The great majority of features used for classification are domain- and language-independent. We demonstrate two ways of using this method of linguistically motivated selection of anaphor-antecedent pairs. First, the amount of training examples stored in memory is reduced. We find that for coreference resolution of definite descriptions and names, the amount of training data can thereby be reduced with only a minor loss in performance, but for pronoun resolution there is a negative effect. Second, selection can be used for improving on coreference resolution results. This is the first step in our hybrid approach to coreference resolution, where the second step is the application of an MBL classifier for determining coreference between the selected pairs. Results indicate that this hybrid approach is advantageous for coreference resolution of definite descriptions and names. For pronoun resolution, there is a negative effect on recall along with a positive effect on precision.
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| 8. |
- Nilsson, Viktor, 1985-
(author)
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Highly Concentrated Electrolytes for Rechargeable Lithium Batteries
- 2020
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Doctoral thesis (other academic/artistic)abstract
- The electrolyte is a crucial part of any lithium battery, strongly affecting longevity and safety. It has to survive rather severe conditions, not the least at the electrode/electrolyte interfaces. Current commercial electrolytes are almost all based on 1 M LiPF6 in a mixture of organic solvents and while these balance the many requirements of the cells, they are volatile and degrade at temperatures above ca. 70°C. The salt could potentially be replaced with e.g. LiTFSI, but dissolution of the Al current collector would be an issue. Replacing the graphite electrode by Li metal, for large gains in energy density, challenges the electrolyte further by exposing it to freshly deposited Li, leading to poor coulombic efficiency and consumption of both Li and electrolyte. Highly concentrated electrolytes (HCEs) have emerged as a possible remedy to all of the above, by a changed solvation structure where all solvent molecules are coordinated to cations – leading to a lowered volatility, a reduced Al dissolution, and higher electrochemical stability, at the expense of higher viscosity and lower ionic conductivity.In this thesis both the fundamentals and various approaches to application of HCEs to lithium batteries are studied. First, LiTFSI–acetonitrile electrolytes of different salt concentrations were studied with respect to electrochemical stability, including chemical analysis of the passivating solid electrolyte interphases (SEIs) on the graphite electrodes. However, some problems with solvent reduction remained, why second, LiTFSI–ethylene carbonate (EC) HCEs were employed vs. Li metal electrodes. Safety was improved by avoiding volatile solvents and compatibility with polymer separators was proven, making the HCE practically useful. Third, the transport properties of HCEs were studied with respect to salt solvation, viscosity and conductivity, and related to the rate performance of battery cells. Finally, LiTFSI–EC based electrolytes were tested vs. high voltage NMC622 electrodes.The overall impressive electrochemical stability improvements shown by HCEs do not generally overcome the inherent properties of the constituent parts, and parasitic reactions ultimately leads to cell failure. Furthermore, improvements in ionic transport can not be expected in most HCEs; on the contrary, the reduced conductivity leads to a lower rate capability. Based on this knowledge, turning to a concept of electrolyte compositions where the inherent drawbacks of HCEs are circumvented leads to surprisingly good electrolytes even for Li metal battery cells, and with additives, Al dissolution can be prevented also when using NMC622 electrodes.
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| 9. |
- Flaquer, Berta
(author)
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Urbanization as Socionatures' Reproduction: from Territories of Extraction
- 2023
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Doctoral thesis (other academic/artistic)abstract
- Through an engagement with the strand of critical urban theory, this dissertation brings the reworkings of Henri Lefebvre’s notion of ‘planetary urbanisation’ into a new synthesis with further inputs from urban political ecology and feminism—towards developing an ecofeminist lens to urbanization. Guided by the hypothesis “urbanization has been historically sustained through the patriarchal domination of women and nature’s reproduction,” the thesis seeks to critically explore how urbanization processes have historically and multiscalarly recurrently transformed the spatial configurations of reproduction from territories of extraction. It does so by engaging with the long durée historical problematique of the malm territory of extraction, as situated from Swedish Sápmi and through an intersectional ecofeminist approach. From the mid-1500s—Indigenous Sámi, Natures, and the bodies of especially women—have been violently subordinated through patriarchal-colonial-capitalist urbanization processes. Across scales and time, the so-called production has been designed by and for the BWMAh* as extraction. Through the malm territory—within and beyond Sweden—this has taken the form of iron ore mining, but also historically in linkage with other forms (i.e. fur and leather, large-scale reindeer, fishing, agriculture, forestry, coal, hydropower, research and development, tourism, data centres, fertilizers, space industry, dredging, fossil-free steel, or fossil- free hydrogen). Backgrounded and at the basis, however, extraction has been sustained through the violent domination of nature and women’s reproduction, as through the witch-hunts in different forms historically and still ongoing, femicides, the creation of the ideals of ‘the good woman,’ the myth of ‘the strong Sámi women,’ successive scientific revolutions, race biology, genetics, industrial colonialism, or the new green colonialism. Under the current ‘green everything’ transition where once again capital’s project is rearticulating and preparing for the next wave of accumulation underway—through an ever-backgrounded and deeper preceding crisis of reproduction—, it is ever more relevant to question the spatiality of the reproduction processes and the ways in which earlier rearticulations have dominated the reproduction of life in new forms.The malm territory is then synchronically and diachronically mapped yet foregrounding the processes of subordination of nature and women—across scales and time—building up the ‘palimpsests of extraction.’ It uses Corboz’s metaphor of ‘the territory as palimpsest’ and expands its conception as a mapping method beyond cartography, to explore in which ways reproduction relations have been masked and not represented historically. The dissertation then goes on to argue that the existing literature on planetary urbanisation has been giving ontological priority to production, and this has precluded an analysis of the actual reproduction relations that have been at the root sustaining life however subordinated in the complete urbanization of society. By advancing an ecofeminist materialist lens to urbanization that reads—representationally and spatially—grasping the complex specificities through the key moments of rearticulation of patriarchal-colonial-capitalist urbanization—temporally and multiscalarly—in the long durée history of the malm territory yet foregrounding the forms of subordination of nature and women, I focus on the linkages that can be drawn between the relations of domination and alternatively collectively transformed.* BWMAh: Bourgeois White Male Adult heterosexual
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| 10. |
- Gerogianni, Alexandra, 1993-
(author)
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The role of the thromboinflammatory response under hemolytic conditions : pathophysiological mechanisms and therapeutic inhibition
- 2023
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Doctoral thesis (other academic/artistic)abstract
- In blood circulation, the complement and the coagulation cascades, together with platelets and endothelial cells form a complex network of crosstalk. When dysregulated, these interactions can lead to inflammation in combination with thrombosis (thromboinflammation) and the manifestation of pathophysiological complications. As complement activation and thromboinflammation are often associated with intravascular hemolysis, e.g., sickle cell disease (SCD), we aimed to study these reactions in relation to heme, a product of hemolysis. Furthermore, our goal was to evaluate whether exposure to biomaterials results in hemolysis-induced thromboinflammation, and to examine the potential of complement inhibition.Our findings show that heme could lead to a significant thromboinflammatory response in our in vitro whole blood model, as seen by complement-, cell- and coagulation- activation, as well as increased cytokine secretion. Inflammation, including complement activation, was also linked with increased heme concentrations in vivo in hemolytic disease in SCD patients. The mechanism of action was attributed to uncontrolled alternative pathway (AP) activation, as heme was shown to bind and inhibit the main AP regulator, factor I, resulting in increased concentrations of fluid phase and surface-bound C3b.Moreover, administration of iron oxide nanoparticles (IONPs) in vitro and implantation of left ventricular assist device (LVAD) in vivo were monitored and correlated with increased hemolytic, e.g., heme, and thromboinflammatory markers, e.g., complement-, endothelial cell- and platelet- activation. Targeting complement components C5 and C3 in vitro was shown overall beneficial in the presence of heme or IONPs respectively. In our settings, the majority of the thromboinflammatory markers measured were successfully attenuated, indicating that complement fuels this response.In conclusion, the results in this thesis stress that heme-induced complement activation is an important player in thromboinflammation. In addition, we propose that complement inhibition can be used as a therapeutic approach in hemolytic conditions and as a strategy to enhance biomaterials’ biocompatibility.
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