Hemos analizado el efecto de la IL-12 sobre la enfermedad autoinmune en un modelo murino semialogénico de enfermedad crónica de injerto contra el huésped (EICHc) inducida en ratones (Balb/c X A/J) F1 (CAF1) inyectados por vía intraperitoneal con células semialogénicas de la cepa parental BALB/c. La IL-12 fue administrada 1 h antes del transplante de células semialogénicas siguiendo dos protocolos diferentes de administración: (a) inyectando 2 g de rmIL-12 (IL-12 murina recombinante) por ratón antes de la primera inyección de células semialogénicas; o (b) inyectando los 2 g de rmIL-12 fraccionada en 5 días. Se produjo una respuesta Th1 pero no apareció la enfermedad aguda de la EICH a pesar de las diferencias en clase I y II de antígenos del complejo mayor de histocompatibilidad (MHC) entre donante y receptor. Cuatro días después de la transferencia de células semialogénicas, los ratones tratados con rmIL-12 mostraron una marcada reducción en el porcentaje de células B comparado con animales control CAF1 y ratones con EICH CAF1+BALB/c. Después de 5-6 meses de seguimiento de la enfermedad, el quimerismo de células donantes incrementó significativamente en bazo (70  31 vs 43  31 %) y en timo. La citometría de flujo de esplenocitos demostró que el quimerismo de células donante estaba constituido por linfocitos T CD4, T CD8 y B y los porcentajes fueron más elevados en animales inyectados con IL-12. Además, el 100% de linfocitos T CD8 fueron de origen donante en los animales con EICH tratados con IL-12 y el 50 % fueron de origen donante en los animales con EICH no tratados. Los resultados demostraron que: (1) La IL-12 probablemente juega un papel en el mecanismo del quimerismo de células donantes, probablemente producido por mecanismos mediados por CTL donantes anti-huésped; (2) no induce la enfermedad aguda de ICH a pesar de las diferencias en clase I y II de moléculas del MHC; (3) la IL-12 no muestra ningún efecto sobre signos clínicos de la enfermedad AR-like desarrollada en este modelo de EICH aunque los signos subclínicos histológicos fueron menos frecuentes, y no se detectó glomerulonefritis en ratones con EICH tratados con IL-12. / We analyzed the IL-12 effect in an autoimmune disease induced in a semiallogenic murine model of graft-vs-host disease (GVHD) Balb/c semiallogenic lymphoid cells i.v. infected in hybrid mice (Balb/c x A/J) F1 (CAF). IL-12 was administered 1 h before cell transplantation following two different protocols: (a) injecting 2 microg of mrIL-12 (murine recombinant IL-12) per mouse before the first semiallogenic cell injection; or (b) injecting the 2 microg of mrIL-12 fractionated in 5 days. ATh1 response was produced but an acute GVHD did not appear although differences in class I and II major histocompatibility complex (MHC) antigens were present. Four days after the semiallogenic cell transfer, IL-12 treated mice showed a marked reduction in the percent of spleen B cells compared with CAF1 control and CAF1 + Balb/c GVHD mice. After 5-6 months of follow-up, the donor cell chimerism increased significantly in spleen (70 +/- 31 vs. 43 +/- 31%) and in thymus. Flow cytometry of spleen lymphocytes demonstrated that donor chimerism was made up of TCD4, TCD8 and B lymphocytes and was higher in animals injected with IL-12. Moreover, CD8 T lymphocytes were 100% donor origin in the IL-12-injected group of GVHD animals and 50% origin in the IL-12-non-injected CAF1 + Balb/c group of animals. This paper shows that: (1) IL- 12 may play a role in the mechanisms of donor cell engraftment, probably produced by a CTL donor anti-host mechanism; (2) no acute GVHD was induced in spite of class I and II MHC differences; (3) IL-12 did not show any effect on the AR-like clinical signs of disease developed in this model of GVHD although histological subclinical signs were less frequent, and no glomerulonephritis was detected in the IL-12-treated GVHD mice.

Sedan år 1965 har Sverige haft koncernbidragsregler som har till syfte att åstadkomma resultatutjämning inom en koncern. Anledningen till att reglerna finns är att beskattningen inte ska styra valet av organisationsform. Beskattningen ska inte variera beroende på om verksamheten bedrivs som en koncern eller som ett enda bolag. De svenska reglerna i 35 kap. IL förutsätter dock att mottagaren av koncernbidraget beskattas i Sverige, vilket gör att reglerna enligt uppsatsen slutsats anses strida mot EU-rätten. Koncernavdragsreglerna i 35 a kap. IL infördes 1 juli 2010 som ett komplement till de redan befintliga koncernbidragsreglerna. Koncernavdragsreglerna har till syfte att under vissa förutsättningar tillåta gränsöverskridande förlustutjämning. Reglerna har tillkommit med anledning av EU- rättens krav på en fri inre marknad. Det ska vara möjligt för företagen att fritt etablera sig i andra EU- länder utan riskera att diskrimineras i jämförelse med andra företag det berörda landet. Kravet på en fri inre marknad kan dock inte upprätthållas strikt eftersom medlemsländerna också har ett motstridigt intresse som är att allokera intäkter till ländernas egna skattebas. Om en fri inre marknad skulle upprätthållas strikt skulle det leda till skatteplanering av företagen där beskattningen skulle förläggas i de länder med förmånligast beskattning. EU har med den bakgrunden accepterat vissa rättfärdigandegrunder där en diskriminering är tillåten. Det gäller därmed för EU:s medlemsländer att balansera de båda intressena i sin lagstiftning. Lagstiftningen ska bidra till att en fri inre marknad upprätthålls men samtidigt inte göra lagstiftningen för generös då det skulle äventyra en välvägd fördelning av beskattningsrätten. Balansgången mellan dessa två intressen har gjort att utformningen av koncernavdragsreglerna i 35 a kap. IL har varit ett stort problem. Uppfyller då de nya reglerna om koncernavdrag kraven på gränsöverskridande förlustutjämning eller kan de fortfarande anses vara fördragsstridiga? Jag kan redan nu nämna att jag har kommit fram till att de nya reglerna i 35 a kap. IL om gränsöverskridande förlustutjämning fortfarande kan anses strida mot EU- rättens krav på en fri inre marknad. Slutsatserna har dragits utifrån det öppna kritiserandet i propositionen, kommentarer från doktrin samt egna åsikter.

<p>Naturally occurring CD4+CD25+Foxp3+ regulatory T cells (TReg) are a cell lineage that develops in the thymus and exits to the periphery, where they represent 5-10% of the peripheral CD4+ T cell population. Phenotypically, they are characterized by the expression of the cell surface markers CD25, as known as the IL-2 receptor alpha chain, glucocorticoid-induced tumor necrosis factor receptor (GITR), and cytotoxic T-lymphocyte antigen-4 (CTLA-4), as well as forkhead box P3 (Foxp3), a transcription factor considered to be the most specific TReg marker. Functionally, TReg cells are defined by their ability to suppress the activation of multiple cell types including CD4+ and CD8+ T cells, B cells, natural killer (NK) cells, and dendritic cells (DCs). Suppression can be achieved by the production of immunosuppressive cytokines or direct cell-to-cell contact, with these mechanisms directly affecting suppressed cells or indirectly affecting them by modulating antigen presenting cells (APCs). The suppressive abilities of TReg cells are crucial in maintaining dominant tolerance--the active, trans-acting suppression of the immune system for the prevention of autoimmune diseases. In addition to preventing autoimmune diseases, studies have also demonstrated critical roles for TReg cells in down-modulating anti-tumor immunity, suppressing allergic diseases, such as asthma, and achieving transplant tolerance. Recent studies have also demonstrated roles for TReg cells during pathogen infection, which will be the focus of this thesis.</p><p>Studies examining TReg cells during infection have largely focused on chronic infection models. These studies have shown that TReg cells can affect responses to pathogens in various ways that can be beneficial or detrimental for either the host or the invading pathogen. In some infections, TReg cells downregulation effector responses, which can lead to pathogen persistence and, in some cases, concomitant immunity. TReg cell-mediated suppression can also reduce immunopathology at sites of infection, which can occur as a result of a vigorous anti-pathogen immune response. </p><p>In contrast to chronic infection, how TReg cells behave and function following acute infections remains largely unknown as, to date, very few studies have been conducted. Current work with acute infection models has indicated that TReg cells affect immune responses in some acute infection models, but not in all. Furthermore, the results of these studies have implicated that current approaches to examine TReg cells during acute infection by depleting the total TReg cell repertoire, as opposed to targeting pathogen-specific TReg cells, may not be ideal. Finally, it is unclear what happens to activated TReg cells following the resolution of infection. </p><p>Due to the lack of knowledge about the role of pathogen-specific TReg cells during acute infection, we sought to employ a different approach to address some of the outstanding questions in the field. Here, we utilized CD4+ non-TReg and TReg cells from T cell receptor (TCR) transgenic mice that recognize a pathogen-specific epitope found in three different models of acute viral infection: recombinant vaccinia virus, recombinant adenovirus, and influenza. Using this model system, we were able to track pathogen-specific TReg cells following acute viral infection to determine their kinetics during the course of infection, as well as their influence on CD4+ non-TReg cells during different times after infection. We also employed major histocompatibility complex (MHC) Class II tetramer technology to track the fate of endogenous pathogen-specific TReg cells following infection with influenza. </p><p>Using these models systems, we show that pathogen-specific TReg cells can be activated and expand upon acute viral infections in vivo. The activated TReg cells then contract to form a "memory" pool after resolution of the infection. These "memory" TReg cells expand rapidly upon a secondary challenge, secrete large amounts of IL-10, and suppress excessive immunopathology, which is elicited by the expansion of non-TReg cells, via an IL-10-dependent mechanism. The work presented in this thesis reveals a previously unknown "memory" TReg cell population that develops after acute viral infections and may help design effective strategies to circumvent excessive immunopathology.</p> / Dissertation

Pseudomonas aeruginosa-induced pneumonia is serious problem that results in severe inflammation response and high mortality in the host. Interleukin-1£] (IL-1£]) is one of the major extracellular proinflammatory cytokines thought to be involved in many acute and chronic lung diseases. To investigate the role of caspase-1, IL-1£] and macrophage inflammatory protein-2 (MIP-2) in P. aeruginosa pneumonia induced lung damage, C57BL/6 (WT) and CASP-1-/- mice were subjected to pneumonia induced by intratracheal injection of P. aeruginosa. The lung permeability, bacterial content in blood and lung, lung myeloperoxidase (MPO) activity, total cell counts and protein in bronchoalveolar lavage fluid (BALF), NF-£eB activation as well as expression of IL-1£] and MIP-2 were assayed at 8 hr after P. aeruginosa injection. The IL-1£] inhibitor, anakinra, was also used to evaluate the role of IL-1£]. P. aeruginosa injection increased the lung permeability, lung MPO activity, bacterial counts in blood, total cell counts and protein in BALF, NF-£eB activation and expression of IL-1£] and MIP-2 in WT mice; and these increases were all decreased by administration of anakinra in WT mice or in CASP-1-/- mice. Furthermore, the lung MPO activity, total protein in BALF and expression of IL-1£] and MIP-2 were decreased in CASP-1-/- ¡÷ WT but not in WT ¡÷ CASP-1-/- chimeric mice, suggesting that pneumonia induced lung damage and IL-1£] and MIP-2 expressions depend on caspase-1 signaling of the resident cells.

Grouper (Epinephelus coioides) is one of the important farmed fish in the southern Taiwan. However, grouper aquaculture in Taiwan has a serious problem of infection, especially in grouper larvae breeding stage. The infection resulted in very high mortality, which causes massive economic loss. Therefore, early detecting the presence of pathogen is critical for preventing epidemic outbreak. Interleukin-1 (IL-1) is one of proinflammatory cytokines that form a feedback control loop with anti-inflammatory cytokines to maintain the homeostasis of host immune response. The increase of IL-1 expression could be an indicator of pathogenic insult. In this study, total RNA of Epinephelus coioides fertilized egg was extracted for reverse transcription-polymerase chain reaction (RT-PCR) to amplify cDNA of IL -1£]. The cDNA amplified was then cloned into pGEX4T-3 for the expression and purification of GST-IL-1£] fusion protein. GST-IL-1£] fusion protein purified was then used to immunize New Zealand white rabbit for generation of antiserum against IL-1£]. Western blot result confirmed the specificity of antiserum as the immune serum, but not the preimmune serum, detected the immunogen GST-IL-1s. Further experiments using live Epinephelus coioides injected with or without lipopolysarcharides (LPS) further confirmed that this antiserum could detect a massive increase of IL-1£] protein after the injection of LPS in either protein lysate by western blotting or in frozen tissue section of head kidney by immunohistochemistry. In summary, we successfully generated a rabbit specific antiserum against IL-1£] of Epinephelus coioides , which could be a useful reagent for future analysis of fish immune response upon pathogen infection.

Hepatic fibrosis represents a process of healing and scarring in response to chronic liver injury. Interleukin-10 (IL-10) is a cytokine that downregulates the proinflammatory response and has a modulatory effect on hepatic fibrogenesis. The aim of this study was to investigate whether IL-10 gene therapy possesses anti-hepatic fibrogenesis in mice. Liver fibrosis was induced by long-term thioacetamide administration in mice. Human IL-10 expression plasmid was delivered via electroporation after liver fibrosis established. IL-10 gene therapy reversed hepatic fibrosis and prevented cell apoptosis in a thioacetamide-treated liver. RT-PCR revealed IL-10 gene therapy could reduce liver transforming growth factor-£]1¡]TGF-£]1¡^, tumor necrosis factor-£\¡]TNF-£\¡^, collagen £\1, cell adhesion molecule, and tissue inhibitors of metalloproteinase¡]TIMPs¡^mRNA upregulation. Following gene transfer, the activation of £\-smooth muscle actin¡]£\-SMA¡^and cyclooxygenase-2¡]COX-2¡^were significantly attenuated. In brief, electroporative IL-10 gene therapy might be an effective therapeutic reagent for liver fibrosis with potential future clinical applications.

ABSTRACT Background: Palindromic rheumatism (PR) is the rare disease that generally occurs with multiple recurring attacks of painful inflammation affecting joints and adjacent tissues. The thesis attempts to characterize the association in 10 instances of the single nucleotide polymorphisms (SNPs) of tumor necrosis factor genes (TNF-£\, TNFRSF1A and TNFRSF1B), Interleukin-1£] genes and Palindromic rheumatism (PR). Methods: The genetic polymorphisms of TNF-£\, TNFR£L, TNFR£S and IL-1£] genes cluster were investigated among 56 PR patients identified from the Kaohsiung Veterans General Hospital (VGHKS, Kaohsiung, Taiwan) and compared with one hundred healthy subjects. The genotypes for ten SNPs in the TNF-£\, TNFRSF1A, TNFRSF1B and IL-1£] genes among these 156 individuals were examined. Results: Experiments indicate significant count of the TNFRSF1A+36 AG genotype in PR patients (OR=4.8, 95%CI=1.8-13.0, p=0.002) and TNFRSF1A+36G allele (OR=3.94, 95%CI=1.59-9.79, p=0.003).The results also have remarkable correlation with TNFRSF1B haplotype +676/+1663 T/A (OR=2.12, CI=1.2-3.8, p=0.010). However, on significant differences were found for all the TNF-£\and IL-1£]polymorphisms. Conclusions: Genetic polymorphisms in TNF-£\ receptors are associated with susceptibility and severity of the inflammatory response in the PR patients.

Previous studies show that coronary artery disease (CAD) is a multi-factors and chronic inflammatory disease, and is associated with lipid metabolism. IL-1ra is a naturally occurring anti-inflammatory molecules that block the action of IL-1. However, little is known about the imbalance between IL-1ra and inflammatory mediators in CAD. We attempted to investigate the relationships between inflammatory mediators and serum IL-1ra levels in patients with CAD. In 95 patients with angiographically defined CAD, and 70 healthy controls were studied in a case-control manner. Serum levels of cytokines and the risk factor of CAD were examined. Polymorphisms for IL-1ra gene were detected by PCR, and genotypes and allelic frequencies in both groups were compared. Our major finding include: (1) The risk factors such as elevated BMI, systolic BP, smoking, hypertension, blood glucose, and TG was more frequently found in the CAD group than the control group ( p < 0.001). However, the HDL-C and bilirubin were significantly higher in control group than the CAD group. (2) The relative risk of those in the highest quartile of ratio of LDL-C to HDL-C, TC to HDL-C, and TG to HDL-C were significantly elevated. ( OR = 2.98, p < 0.01; OR = 5.31, p <0.001; OR = 8.43, p < 0.001 respectively) (3) Five different inflammatory markers were significantly elevated including IL-1ra, hs-CRP, IL-6, leukocyte count, and neutrophil percentage between healthy controls and CAD patients. ( p < 0.01) (4) Levels of IL-1ra and other variables such as blood glucose, BMI, TG, IL-6, hs-CRP, and leukocyte count has significantly correlated, and were inversed correlation in bilirubin, and HDL-C in all study subjects. ( p < 0.01) (5) In the multiple logistic regression analysis, adjustment was made for variables. The relative risk of CAD for the highest quartile of IL-1ra, as compared with the lowest quartile, had an Odds ratio 2.57 ( 95% confidence intervals, 1.12 - 5.91, p = 0.026 ) increase in risk for CAD. (6) Similar results were obtained hs-CRP, IL-6 in the highest quartile were increase risk for future CAD. ( OR = 5.86 and 5.79 respectively; p < 0.001) (7) The join effect cytokines of hs-CRP, IL-6, IL-1ra concentrations may play important role in CAD risk. ( OR = 10.19, p < 0.001 ) (8) In addition, IL-1ra allele 2 genotype and allelic frequencies were no significant association with increase in IL-1ra with CAD. In conclusion, we find a significant association of elevated IL-1ra levels in the patients with CAD. Thus, these results support the hypothesis that inflammation, anti-inflammation cytokines and lipoprotein metabolism provide a useful marker for predicting the development of CAD events.

<p>Vid beskattning finns det två sidor med olika intressen. Dels den enskilde individen som bevakar sitt intresse av att minimera sin skattebelastning och dels staten med ett intresse för skyddet av skattebasen. Vid en flytt från Sverige kan obegränsad skattskyldighet aktualiseras på grund av att den skattskyldige har väsentlig anknytning till Sverige.</p><p>I svensk skatterätt har legalitetsprincipen, ingen skatt utan lag, en stark ställning. Detta ställer krav på lagstiftningen att den skall uppfylla ett visst mått av förutsägbarhet. Den skattskyldige skall kunna förutse konsekvenserna av sitt handlande. 3:7 IL innehåller tio anknytningsmoment som vid en sammanvägd bedömning skall ligga till grund för graden av skattskyldighet. Ur lagtexten går det dock inte att utläsa vad dessa anknytningsmoment i praktiken innebär. Ledning för ett beslut bör då sökas i förarbete, praxis och doktrin.</p><p>Praxis på området om väsentlig anknytning är inte enhetlig och gällande en del anknytningsmoment följer inte praxis de kriterier förarbetena stadgat som norm för bedömning. Detta gör att rättssäkerheten såväl som effektiviteten gällande lagrummet 3:7 IL som det är utformat idag kan ifrågasättas. Vidare saknar många domar från Regeringsrätten motiveringar till varför utgången i målet blev som den blev, vilket ytterligare bidrar till en osäkerhet och otydlighet på området.</p><p>Någon lagstadgad rangordning av de tio anknytningsmomenten finns inte utan det är genom praxis viktningen av anknytningsmomenten har skett. Ett av de viktigaste anknytningsmoment är<em> bostad inrättad för åretruntbruk</em>. Praxis på området har tenderat att förskjutas från att objektivt värdera standard och läge på fastigheten till att subjektivt bedöma hur fastigheten nyttjas. Detta innebär att fritidsbostäder, oavsett läge och standard, inte medför väsentlig anknytning, medans fall där den skattskyldige behåller en permanentbostad alltid betraktas ha väsentlig anknytning. <em>Familj</em> utgör ett starkt skäl till att obegränsad skattskyldighet består, särskilt om familjen kvarstannar. Regelbundna besök till Sverige ses som en indikation på att den skattskyldige inte har för avsikt att bryta banden med Sverige. Vidare utgör <em>ekonomisk engagerad här genom att inneha tillgångar som, direkt eller indirekt, ger honom väsentligt inflytande </em>och<em> om han bedriver näringsverksamhet här</em> starka grunder för fortsatt obegränsad skattskyldighet i Sverige trots en flytt härifrån. Gränsdragningen för storleken på tillgångar i relation till skattskyldighet är svårbedömd. Generellt kan sägas att ett innehav som inte överstiger 10 % av tillfångarna i ett fåmansföretag bör inte medföra väsentlig anknytning medans ett innehav över 15 % gör det. Gällande anknytningsmomentet <em>om han bedriver näringsverksamhet här</em> antas vanligen att den skattskyldige har så täta kontakter med Sverige att det medför väsentlig anknytning.</p><p>För att öka både rättssäkerheten och effektiviteten kan förespråkas att domstolen blir tydligare i sin utformning av domar och beslut. Likaså att domarna motiveras väl och att det klart och tydligt framgår varför beslutet blev som det blev. I ett steg i att förbättra både rättssäkerheten och effektiviteten kan förslagsvis ett införande av en ”specialdomstol”, vars ansvarsområde bland annat berör frågor runt väsentlig anknytning, motiveras. Ur ett effektivitetsperspektiv skulle detta troligen förkorta handläggningstider men även höja kompetensen hos domstolen och på så sätt öka förutsägbarheten, minska godtycklighet på området och då minska antalet situationer som kommer till prövning i domstolen.</p>

Elevated levels of reactive oxygen species (ROS) cause or aggravate a variety of pathological conditions such as cardiovascular disease, cancer and rheumatoid arthritis. Despite known links between oxidative stress and disease, years of clinical studies have failed to show clear benefits of antioxidant therapy. It is now recognized that ROS such as hydrogen peroxide can act as signaling molecules and are required for physiological functioning of a number of signaling pathways. Therefore, a mechanistic basis of ROS-mediated regulation of cell signaling must be established to enable rational design of antioxidant-based therapies. The challenges in quantification of transient changes mediated by ROS during cell signaling have impeded investigation of redox-regulated signaling. In the present work, computational modeling is used to circumvent these technical challenges and to test competing hypotheses of redox regulation. Using a quantitative, systems level approach to study interactions between ROS dependent and independent regulatory mechanisms, the most comprehensive model of the IL-4 signaling pathway to date has been developed and validated with experimental data. The model is capable of predicting kinase phosphorylation dynamics under new oxidative conditions, and our analyses suggest that reversible oxidation of tyrosine phosphatases is the primary mechanism of redox regulation in this pathway. Additional computational methods have been developed to study ROS as mediators of crosstalk between signaling pathways, to optimize model parameters, and to interrogate model dynamics for the purpose of model selection. Collectively, these modeling tools provide a new systems-level perspective for investigating reversible protein oxidation as a means of control over cellular signal transduction.