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Submitted by LORENZO GARRIDO TEIXEIRA MARTINI SEGABINAZZI null (lorenzosegabinazzi@unesp.com.br) on 2016-11-11T16:37:12Z No. of bitstreams: 1 Dissertação de mestrado Lorenzo FINAL.pdf: 2227722 bytes, checksum: 5a7e643913e44a96155c8fd80a611918 (MD5) / Approved for entry into archive by LUIZA DE MENEZES ROMANETTO null (luizaromanetto@hotmail.com) on 2016-11-16T16:09:30Z (GMT) No. of bitstreams: 1 segabinazzi_lgtm_me_bot.pdf: 2227722 bytes, checksum: 5a7e643913e44a96155c8fd80a611918 (MD5) / Made available in DSpace on 2016-11-16T16:09:31Z (GMT). No. of bitstreams: 1 segabinazzi_lgtm_me_bot.pdf: 2227722 bytes, checksum: 5a7e643913e44a96155c8fd80a611918 (MD5) Previous issue date: 2016-09-14 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / A endometrite persistente pós-cobertura (EPPC) é a principal causa de redução da fertilidade nas éguas, causando impactos importantes dentro do mercado do cavalo. Os tratamentos comumente utilizados para EPPC visam apenas minimizar os fatores predisponentes a sua instalação, não atuando diretamente no processo inflamatório. Com o intuito de reduzir a resposta inflamatória, estudos recentes têm demonstrado um aumento da fertilidade de animais acometidos por EPPC, quando se utiliza agentes imunomoduladores. O PRP é modulador da resposta inflamatória que está sendo largamente utilizado na medicina veterinária. Este concentrado de plaquetas contém diversos fatores de crescimento que atuam diretamente nos mediadores inflamatórios, reduzindo o processo e promovendo reparação tecidual. O PRP é benéfico no tratamento de inflamações tendíneas e osteoarticulares, modulando a inflamação e acelerando a regeneração do tecido lesionado. Mais recentemente alguns pesquisadores demonstraram o efeito benéfico do PRP em tratamentos intrauterinos de éguas. Desta forma o presente estudo tem por objetivo revisar os aspectos relacionados a EPPC assim como ao PRP e seu mecanismo de ação. / The persistent breeding-induced endometritis (PBIE) is the main cause of decrease fertility in the horses, thereby causing significant impact in the horse’s market. The treatments commonly used for PBIE view only minimize the predisposing factors and do not act directly in the inflammatory process. Aiming to reduce the inflammatory response, recent studies have shown an increase in fertility of animals with PBIE when used immunomodulatory agents. A modulator of the inflammatory response that has been largely used in veterinary medicine is the platelet-rich plasma (PRP). This platelet concentrate contains many growth factors which act directly on inflammatory mediators, reducing process and promoting tissue repair. Several studies have shown that PRP is beneficial in the treatment of osteoarticular and tendon inflammations, modulating inflammation and accelerates the regeneration of injured tissue. More recently some researchers have demonstrated the beneficial effect of PRP in intrauterine treatment of mares. Thus, the present study aimed to do a literature review on the aspects related to PBIE, as well as the PRP and its mechanism of action. / FAPESP: 2015/00150-8

Os β- glucanos são polissacarídeos que possuem atividade imunomoduladora. A colibacilose é uma das principais doenças da avicultura, pois causa grandes perdas econômicas. Diante da necessidade de se estabelecer a ação do β-glucano solúvel nas aves, foram realizados dois experimentos. Um para se avaliar os efeitos do β-glucano solúvel, administrado por diferentes vias, na produção de anticorpos séricos IgA e IgG, nos parâmetros zootécnicos e outro na infecção por Escherichia coli (EC). Os experimentos foram realizados ao mesmo tempo, em galpões diferentes, sendo alojados 160 frangos de corte da linhagem Ross (Aviagen®), do primeiro ao 42o dia de idade. β-glucano solúvel foi administrado no primeiro dia, na dose de 215µg/ave, nas seguintes vias de administração: subcutânea (SC), ocular (OC) e oral (VO). O controle negativo foi o mesmo para os dois experimentos. Os tratamentos do Experimento 1 (Vias de Administração) foram: T2(SC), T3(OC) e T4(VO). O experimento onde as aves foram desafiadas por Escherichia coli foi o Experimento 2 (β-glucano e Escherichia coli). As mesmas vias de inoculação foram também estudadas nestes tratamentos, assim: T5 (controle positivo) foi desafiado e não recebeu o β-glucano solúvel, T6(SC+EC), T7(OC+EC) e T8(VO+EC). O desafio foi realizado com E. coli sorotipo O119, por via saco aéreo torácico direito aos 14 dias de idade, sendo que cada ave recebeu 4x106 UFC. As aves foram pesadas semanalmente para a análise dos parâmetros zootécnicos e toda ração adicionada foi pesada. Aos 28, 35 e 42 dias foi colhido sangue de todas as aves para se determinar por ensaio imunoenzimático indireto (ELISA indireto) as concentrações séricas de IgG e IgA totais. Nas aves infectadas foi determinado o escore de lesão, analisando a intensidade das lesões em sacos aéreos, fígado e coração. No Experimento 1 (Vias de Inoculação) os resultados mostraram concentrações séricas de IgA nas aves do T4(OC) menores que nas aves dos demais tratamentos. Em relação a IgG as aves do T4(VO) apresentaram concentrações maiores que aquelas dos demais tratamentos. No Experimento 2 (β-glucano e Escherichia coli) as aves do T7(OC+EC) tiveram as menores concentrações de IgA e apresentaram a melhor viabilidade dentre as aves dos grupos desafiados. As aves do T6(SC+EC) apresentaram concentrações maiores de IgG aos 35 e 42 dias. Quanto ao escore de lesão houve redução no grau de lesão em todos os grupos que receberam o β- glucano solúvel, exceto no saco aéreo torácico direito. Sugere-se que o uso do β- glucano solúvel possa reduzir as lesões causadas por Escherichia coli. A administração por via ocular levou a melhor viabilidade nas aves desafiadas, apesar de ter menor produção de anticorpos IgA tanto nas aves infectadas como nas aves não desafiadas. Serão necessários mais estudos para se estabelecer melhor as ações imunomoduladoras do β-glucano nas aves. / The β-glucans are polysaccharides that possess immunomodulatory activity. The colibacillosis is a major disease of poultry; it contributes to great economic losses. Faced with the need to establish the action of soluble β-glucan in birds, two experiments were conducted. One to evaluate the effects of soluble β-glucan administered by different routes, in the production of serum IgA and IgG antibodies in zootechnical parameters and other infection in Escherichia coli (EC). The experiments were performed simultaneously in different warehouses, being housed 160 broilers Ross (Aviagen ®) from first to 42nd days of age. β-glucano soluble was administered on the first day, the dose of 215µg/bird on the following routes of administration: subcutaneous (SC), ocular (OC) and oral (PO). The negative control was the same for both experiments. The treatments of the experiment called Experiment 1 (Routes of Administration) were: T2 (SC), T3(OC) and T4(VO). The experiment where birds were challenged by Escherichia coli was termed Experiment 2 (β-glucan e Escherichia coli). The same routes of inoculation were also studied in these treatments, thus: T5 (positive control), was challenged and did not receive soluble β-glucan, T6(SC+EC), T7(OC+EC) and T8(VO+EC). The challenge was performed with E. coli serotype O119, through the right thoracic air sac at 14 days of age, each bird was given 4x106 CFU. The birds were weighed weekly for analysis of zootechnical parameters ration added and the whole was weighed. At 28, 35 and 42 days blood was collected from all birds to determine by Enzyme linked immunosorbent assay (ELISA) serum concentrations of total IgG and IgA. In birds infected was determined the injury score by analyzing the intensity of the lesions in air sacs, liver and heart. In the Experiment 1 (Routes of Inoculation) the results showed serum IgA in birds of T4(OC) smaller than the other treatments in birds. Regarding IgG the birds of T4(VO) had levels greater than those of other treatments. In Experiment 2 (β-glucan and Escherichia coli) birds of T7(OC+EC) had the lowest concentrations of IgA and showed the best viability among groups of birds challenged. Birds of T6(SC+EC) showed higher concentrations of IgG at 35 and 42 days. As for the injury score decreased the degree of injury in all groups receiving β- glucan soluble except in right thoracic air sac. It is suggested that the use of β-glucan soluble can reduce injuries caused by Escherichia coli. The administration by ocular route led to better viability challenged birds, despite having lower production of IgA in both the infected birds as the birds not challenged. Further studies are needed to better establish the immunomodulatory actions of β-glucan in birds.

L’irradiation cérébrale thérapeutique comporte un risque de neurotoxicité tardive irréversible en partie lié à l’effet de l’irradiation sur le compartiment vasculaire cérébrale. Les péricytes et les communications entre les péricytes et les cellules endothéliales jouent un rôle majeur dans la stabilisation des vaisseaux, dans la formation et la régulation de la barrière hématoencéphalique et dans le contrôle du flux sanguin cérébral. Nous montrons, dans un modèle murin d’irradiation cérébrale, que les péricytes sont une cible précoce de l’irradiation cérébrale. Après irradiation, la morphologie des péricytes est modifiée et des marqueurs d’activations du péricytes sont surexprimés. En conséquence, la communication entre péricyte et cellule endothéliale est rompue, ce qui se traduit par une diminution de la capacité du péricyte à induire une constriction vasculaire après stimulation électrique. De façon concomitante, la perméabilité de la barrière hémato-encéphalique est anormalement augmentée après irradiation. Un traitement par thalidomide, administré dans la semaine précédant et suivant l’irradiation, prévient les conséquences de l’irradiation sur les péricytes. Les communications entre péricytes et cellules endothéliales sont maintenues ainsi que les fonctions contractiles des péricytes. L’imperméabilité de la barrière hématoencéphalique est également préservée. La voie de signalisation PDGF-β/PDGFR-β essentielle au recrutement des péricytes par les cellules endothéliales, est, au moins partiellement, impliquée dans l’effet protecteur de la thalidomide. Des études supplémentaires sont nécessaires pour définir les mécanismes sous tendant l’effet de l’irradiation sur les péricytes ainsi que l’effet protecteur de la thalidomide. Nous avons en parallèle mis au point un modèle murin de lymphome cérébral luciférase positif pour vérifier, dans un premier temps, l’innocuité de l’association de la radiothérapie et de la thalidomide et de ses dérivés de la classe des immunomodulateurs (iMids), le lénalidomide et le pomalidomide. Ces trois molécules ne diminuent pas l’effet antitumoral de la radiothérapie, mais l’association de radiothérapie et de pomalidomide est synergique sur la décroissance tumorale mesurée par l’évolution des courbes de bioluminescence. Le concept de normalisation de la vascularisation tumorale fait référence aux molécules capables, non pas de faire régresser les vaisseaux tumoraux anormaux, mais de les « normaliser » pour améliorer, d’une part, la disponibilité des chimiothérapies au sein de la tumeur, et d’autre part, l’oxygénation tumorale pour accroitre l’efficacité de la radiothérapie. Nos résultats sont en faveur d’un tel effet exercé par le pomalidomide dans notre modèle murin de lymphome cérébral, caractérisé par une infiltration tumorale périvasculaire, une fuite capillaire mais sans néo angiogenèse. Nos travaux fournissent un rationnel biologique à de futurs essais cliniques avec les iMids dans le traitement des lymphomes cérébraux primitifs voire des tumeurs malignes cérébrales. / Therapeutic brain irradiation carries a risk of irreversible delayed neurotoxicity partly due to the effect of irradiation on the cerebral vascular compartment. Pericytes and communication between pericytes and endothelial cells play a major role in vessel stabilization in the formation and regulation of the blood-brain barrier and in the control of cerebral blood flow. We show in a murine model of brain irradiation, that pericytes are a target of early brain irradiation. After irradiation, the morphology of pericytes is altered and markers of activation of pericytes are overexpressed. Consequently, communication between the endothelial cell and pericyte is disrupted , which results in a decreased capacity of pericytes for inducing a vascular constriction after electrical stimulation. Concomitantly, the permeability of the blood - brain barrier is abnormally increased after irradiation. Treatment with thalidomide administered in the week before and after irradiation, prevents the effects of irradiation on pericytes . Communication between pericytes and endothelial cells are maintained as well as the contractile properties of pericytes. The impermeability of the blood brain barrier is also preserved. The PDGF-β/PDGFR-β signaling pathway, which is essential for the recruitment of pericytes by endothelial cells, is at least partially involved in the protective effect of thalidomide. Further studies are needed to define the mechanisms underlying the effect of irradiation on the pericytes and the protective effect of thalidomide. We have, in parallel, developed a model of murine luciferase positive CNS lymphoma to verify first, the safety of the combination of radiotherapy and thalidomide and its derivatives of the class of immunomodulators ( IMiDs ), lenalidomide and pomalidomide. These three molecules do not decrease the antitumor effect of radiotherapy, but the antitumoral effect of the association of radiotherapy and pomalidomide is synergistic. The concept of the normalization of the tumor vascularization refers to molecules capable not to induce regression of the abnormal tumor vessels but to "normalize" the tumoral vasculature in order to improve, on one hand , the availability of chemotherapy in the tumor , and on the other hand, the tumor oxygenation to increase the effectiveness of radiotherapy. Our results are in favor of such an effect exerted by pomalidomide in our murine model of cerebral lymphoma, characterized by perivascular tumor infiltration and capillary leak .Our work provide a biological rational for future clinical trials with IMiDs in the treatment of brain lymphomas or malignant brain tumors.

Os β- glucanos são polissacarídeos que possuem atividade imunomoduladora. A colibacilose é uma das principais doenças da avicultura, pois causa grandes perdas econômicas. Diante da necessidade de se estabelecer a ação do β-glucano solúvel nas aves, foram realizados dois experimentos. Um para se avaliar os efeitos do β-glucano solúvel, administrado por diferentes vias, na produção de anticorpos séricos IgA e IgG, nos parâmetros zootécnicos e outro na infecção por Escherichia coli (EC). Os experimentos foram realizados ao mesmo tempo, em galpões diferentes, sendo alojados 160 frangos de corte da linhagem Ross (Aviagen®), do primeiro ao 42o dia de idade. β-glucano solúvel foi administrado no primeiro dia, na dose de 215µg/ave, nas seguintes vias de administração: subcutânea (SC), ocular (OC) e oral (VO). O controle negativo foi o mesmo para os dois experimentos. Os tratamentos do Experimento 1 (Vias de Administração) foram: T2(SC), T3(OC) e T4(VO). O experimento onde as aves foram desafiadas por Escherichia coli foi o Experimento 2 (β-glucano e Escherichia coli). As mesmas vias de inoculação foram também estudadas nestes tratamentos, assim: T5 (controle positivo) foi desafiado e não recebeu o β-glucano solúvel, T6(SC+EC), T7(OC+EC) e T8(VO+EC). O desafio foi realizado com E. coli sorotipo O119, por via saco aéreo torácico direito aos 14 dias de idade, sendo que cada ave recebeu 4x106 UFC. As aves foram pesadas semanalmente para a análise dos parâmetros zootécnicos e toda ração adicionada foi pesada. Aos 28, 35 e 42 dias foi colhido sangue de todas as aves para se determinar por ensaio imunoenzimático indireto (ELISA indireto) as concentrações séricas de IgG e IgA totais. Nas aves infectadas foi determinado o escore de lesão, analisando a intensidade das lesões em sacos aéreos, fígado e coração. No Experimento 1 (Vias de Inoculação) os resultados mostraram concentrações séricas de IgA nas aves do T4(OC) menores que nas aves dos demais tratamentos. Em relação a IgG as aves do T4(VO) apresentaram concentrações maiores que aquelas dos demais tratamentos. No Experimento 2 (β-glucano e Escherichia coli) as aves do T7(OC+EC) tiveram as menores concentrações de IgA e apresentaram a melhor viabilidade dentre as aves dos grupos desafiados. As aves do T6(SC+EC) apresentaram concentrações maiores de IgG aos 35 e 42 dias. Quanto ao escore de lesão houve redução no grau de lesão em todos os grupos que receberam o β- glucano solúvel, exceto no saco aéreo torácico direito. Sugere-se que o uso do β- glucano solúvel possa reduzir as lesões causadas por Escherichia coli. A administração por via ocular levou a melhor viabilidade nas aves desafiadas, apesar de ter menor produção de anticorpos IgA tanto nas aves infectadas como nas aves não desafiadas. Serão necessários mais estudos para se estabelecer melhor as ações imunomoduladoras do β-glucano nas aves. / The β-glucans are polysaccharides that possess immunomodulatory activity. The colibacillosis is a major disease of poultry; it contributes to great economic losses. Faced with the need to establish the action of soluble β-glucan in birds, two experiments were conducted. One to evaluate the effects of soluble β-glucan administered by different routes, in the production of serum IgA and IgG antibodies in zootechnical parameters and other infection in Escherichia coli (EC). The experiments were performed simultaneously in different warehouses, being housed 160 broilers Ross (Aviagen ®) from first to 42nd days of age. β-glucano soluble was administered on the first day, the dose of 215µg/bird on the following routes of administration: subcutaneous (SC), ocular (OC) and oral (PO). The negative control was the same for both experiments. The treatments of the experiment called Experiment 1 (Routes of Administration) were: T2 (SC), T3(OC) and T4(VO). The experiment where birds were challenged by Escherichia coli was termed Experiment 2 (β-glucan e Escherichia coli). The same routes of inoculation were also studied in these treatments, thus: T5 (positive control), was challenged and did not receive soluble β-glucan, T6(SC+EC), T7(OC+EC) and T8(VO+EC). The challenge was performed with E. coli serotype O119, through the right thoracic air sac at 14 days of age, each bird was given 4x106 CFU. The birds were weighed weekly for analysis of zootechnical parameters ration added and the whole was weighed. At 28, 35 and 42 days blood was collected from all birds to determine by Enzyme linked immunosorbent assay (ELISA) serum concentrations of total IgG and IgA. In birds infected was determined the injury score by analyzing the intensity of the lesions in air sacs, liver and heart. In the Experiment 1 (Routes of Inoculation) the results showed serum IgA in birds of T4(OC) smaller than the other treatments in birds. Regarding IgG the birds of T4(VO) had levels greater than those of other treatments. In Experiment 2 (β-glucan and Escherichia coli) birds of T7(OC+EC) had the lowest concentrations of IgA and showed the best viability among groups of birds challenged. Birds of T6(SC+EC) showed higher concentrations of IgG at 35 and 42 days. As for the injury score decreased the degree of injury in all groups receiving β- glucan soluble except in right thoracic air sac. It is suggested that the use of β-glucan soluble can reduce injuries caused by Escherichia coli. The administration by ocular route led to better viability challenged birds, despite having lower production of IgA in both the infected birds as the birds not challenged. Further studies are needed to better establish the immunomodulatory actions of β-glucan in birds.

Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacology and Toxicology Student: Kristína Hudáková Supervisor: doc. PharmDr. František Trejtnar, CSc. Title of diploma thesis: In vitro evaluation of novel Toll-like receptor ligands I Vaccination against preventable infections prevents millions of deaths each year. Their immunity enhancing activity is strengthened by the presence of vaccine adjuvants. Development of vaccine adjuvants leads to improved safety profile and also can play a vital role in the research of new vaccines against pathogens against which the vaccines currently do not exist. The main aim of this diploma thesis was to verify the ability of rationally developed small molecule ligands to influence Toll-like receptors and thus their potential to be utilized as vaccine adjuvants. The assay was carried out using modified cell lines continually expressing the human TLR4 or TLR8 whose activation leads to production of secreted embryonic alkaline phosphatase. Ten analyzed substances labelled as DM 001 - DM 010 were examined for their agonistic and also antagonistic properties while interacting with the TLRs. Immunomodulatory activity of these tested samples was then determined by quantification of secreted alkaline phosphatase with the help of a colorimetric...

Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacology & Toxicology Student: Vanda Machalová Supervisor: doc. PharmDr. František Trejtnar, CSc. Title of diploma thesis: In vitro evaluation of novel Toll-like receptor ligands II. The aim of this diploma thesis was to test the new substances at Toll-like receptors (TLR), namely TLR4 and TLR8 subtypes, as potential vaccine adjuvants. Adjuvants are required for new subunit vaccines to promote stimulation of a sufficient immune response. Stimulation of TLR receptors is safe and leads to the activation of both innate and adaptive immunity and to subsequent specific immune response. Testing was carried out on cell lines stable co-expressing the respective receptors, with colorimetric activity detection. In this work, both agonist and antagonist activity at TLR4 and TLR8 receptors was investigated and a half maximal effective concentration (EC50) of resiquimod was determined. Based on the results, the respective substances cannot be considered as potential adjuvants to the TLR8 receptor and DM014 exhibits the potential for agonist activity on the TLR4 receptor, whereas the substance DM015 can be used on further investigation of immunosuppression in autoimmune diseases and diseases associated with excessive activation of the...

Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacology and Toxicology Student: Linda Tamášiová Supervisor: doc. PharmDr. František Trejtnar, CSc. Title of diploma thesis: In vitro evaluation of novel Toll-like receptor ligands III Research on new ligands with potential immunomodulatory activity is a tool for the development of new immunological adjuvants for use in vaccines, or as separate immunomodulators in the treatment of various diseases. The aim of this thesis was to analyze the ability of new ligands to stimulate toll-like receptors 4 (TLR4) and to assess the potential for further use of these substances based on established immunomodulatory activity. The analysis was performed on modified TLR4-expressing cell lines whose activation was subsequently detected using a colorimetric-enzymatic reaction. Sixteen new substances were tested for TLR4 receptor activation in comparison with a standard agonist. The results showed a significant TLR4 agonistic activity in several of the test substances, suggesting that they are activating ligands of the receptor tested. However, due to their low solubility, some of these substances are not suitable candidates for further use and testing. Taking all parameters into consideration, among of all of the evaluated substances that...

Bakgrund: Crohns sjukdom är en form av inflammatorisk tarmsjukdom som kännetecknas av kronisk histologisk inflammation, en autoimmun sjukdom som inte är medicinskt botbar men det finns olika medicinska terapier som kan kontrollera symtomen. Det förekommer omväxlande perioder med skov och lugn, operation behövs ibland, medicineringen är konstant och alla dessa faktorer leder till försämrad livskvalitet. Crohns sjukdom skadar ofta ileum men det kan påverka vilket segment av mag-tarmkanalen som helst från mun till perianalt område, lesionerna kan ha formen av fläckar, med vissa delar påverkade medan andra förblir helt normala. Incidensen av Crohns sjukdom i Sverige är mellan 5 och 10 per 100 000 och prevalensen är cirka 200 per 100 000. Medan dödsfallet med Crohns sjukdom minskar, ökar de andra problemen som är förknippade med Crohns sjukdom och detta sätter press på sjukvården. För att hålla sjukdomen under kontroll behöver patienter farmakologisk induktionsbehandling i början av sjukdomen och underhållsbehandling under resten av livet. Just för att behandlingen behövs på lång sikt (livet ut) har det varit absolut nödvändigt att hitta den bästa behandlingsstrategin. Målet med behandlingen är att göra patienten symptomfri med hjälp av medicinering. Mål: Syftet med detta examensarbete var att ta reda på hur tillgängliga biologiska läkemedel ska användas för att få optimal effekt, närmare bestämt om de biologiska läkemedlen ska användas som monoterapi eller i kombination med andra läkemedel vid behandling av Crohns sjukdom. Metod: PubMed-databasen har använts för att söka, välja och analysera fem relevanta randomiserade kontrollerade kliniska prövningar om Crohns sjukdom. Nyckelordet som användes för att hitta alla dessa fem relevanta artiklar var ''Crohn's combination therapy''. Resultat: Resultaten i den första artikeln påpekar att kombinationsbehandling med infliximab + azatioprin hade bättre resultat jämfört med infliximab monoterapi och infliximab monoterapi hade bättre resultat jämfört med azatioprin monoterapi för att uppnå klinisk remission och slemhinneläkning. Den andra artikeln strider mot den första studien och säger att om tillräckliga läkemedelskoncentrationer uppnås och bibehålls genom terapeutisk läkemedelsövervakning med infliximab, kanske kombinationsterapi med tiopuriner inte behövs för att uppnå önskade kliniska resultat. Den tredje artikeln drog slutsatsen att adalimumab var överlägsen tiopuriner i att förhindra endoskopiskt återfall av Crohns sjukdom hos patienter med hög risk för återfall efter operation. Den fjärde artikeln identifierade ingen effektivitetsfördel med kombinationsbehandling med adalimumab + immunmodulatorer jämfört med adalimumab monoterapi. Den femte artikeln drog slutsatsen att vedolizumab i kombination med stabila doser av kortikosteroider inducerade mer effektivt klinisk remission än antingen placebo plus kortikosteroider eller enbart vedolizumab. Slutsatser: En universell slutsats kan inte dras om fördelen med kombinationsbehandling bestående av biologiska och immunsuppressiva läkemedel jämfört med biologisk monoterapi. Olika tillvägagångssätt är nödvändiga för varje enskild patient eftersom medicinen kan vara olika för induktions- och underhållsterapier, sjukdomsvaraktigheten påverkar hur patienten kommer att reagera på mediciner och medicineringen som Crohns sjukdoms-patienter fick tidigare kan också påverka hur patienten kommer att svara på medicinering. / Background: Crohn's disease is a form of inflammatory bowel disease characterized by chronic histological inflammation, an autoimmune disorder that is not medically curable but there are various medical therapies able to control the symptoms. It has periods of exacerbation and calmness, with surgery needed sometimes, the use of medication is constant and all these factors lead to an impaired quality of life. Crohn's disease often damages the ileum but it can affect any segment of the gastrointestinal tract from mouth to perianal area, the lesions may take the form of patches, with some sections affected while others remain perfectly normal. The incidence of Crohn's disease in Sweden is between 5 and 10 per 100,000 and the prevalence is around 200 per 100,000. While the fatal burden is declining, the non-fatal burden is increasing. In order to keep the disease under control, patients require pharmacological induction treatment at the beginning and maintenance treatment for the rest of their life. Because the treatment is needed long-term, finding the best treatment strategy has become imperative. The goal with the therapy is to make the patient symptom-free with the help of medication. Objective: The purpose of this degree project is to find out how biological drugs should be used in order to have optimal effect, more specifically, if the biological medicines should be used as monotherapy or in combination with other medicines in the treatment of Crohn's disease. Method: PubMed database has been used to search, select and analyze five relevant randomized controlled clinical trials about Crohn’s disease. The keyword used to find all these five relevant articles were ''Crohn's combination therapy''. Results: The results in the first article point out that Infliximab + Azathioprine combination therapy had better results compared to infliximab monotherapy and infliximab monotherapy had better results compared with Azathioprine monotherapy in achieving clinical remission and mucosal healing. The second article contradicts the first study and states that if adequate drug concentrations are achieved and maintained through therapeutic drug monitoring with infliximab, combination therapy with thiopurines may not be required to achieve desired clinical results. The third article concluded that adalimumab was superior to thiopurines in preventing endoscopic recurrence of Crohn's disease in patients with high risk of recurrence after surgery. The fourth article identified no efficacy benefit of adalimumab + immunomodulators combination therapy compared to adalimumab monotherapy. The fifth article concluded that vedolizumab in combination with stable doses of corticosteroids induced more efficient clinical remission than either placebo plus corticosteroids or vedolizumab alone. Conclusions: A universal conclusion cannot be drawn regarding the superiority of combination treatment consisting of biological and immunosuppressive medicines compared to biological monotherapy. Different approaches are necessary for every individual patient because the medication can be different for induction and maintenance therapies, the duration of the disease affects how the patient will respond to medication, and the medication that the Crohn’s disease patient took in the past may also affect how the patient will respond to medication.

Peptídeos de defesa do hospedeiro antimicrobiano (PDHA) são componentes do sistema imune inato podem ser expressos constitutivamente ou induzidos por componentes bacterianos, citocinas, ou a combinação de ambos. Estes peptídeos possuem ação direta sobre microrganismo e são potentes imunomoduladores de células de mamíferos. A microplusina é um peptídeo antimicrobiano quelante de cobre, originário do aracnídeo Tick Rhipicephalus (Boophilus) microplus, com massa molecular de 10 204 Da. Este peptídeo possue atividade bacteriostática sobre Micrococcus luteus, mas sem atividade microbicida sobre E. coli e Candida albicans. Neste trabalho foi analisado o efeito direto da microplusina em cultura de macrófagos derivados de medula óssea (MDM) tratados ou não com LPS ou IFN-&#947. Foi demonstrado que este peptídeo não teve efeito citotóxico para macrófagos, não estimulou a síntese de NO e IL-1&#946, mas aumentou a produção de TNF-&#945 e IL-6 após estímulo. O pré-tratamento dos macrófagos com microplusina, antes da adição do LPS, não modificou o nível de nitrito, TNF-&#945 ou IL-6 em resposta ao LPS. Quando os macrófagos foram pré-tratados com microplusina e posteriormente incubados com IFN-&#947 houve aumento significativo da produção de NO e TNF-&#945, confirmando a natureza pró-inflamatória deste peptídeo. Isto indica que a microplusina é um agente imunomodulador da resposta imune inata e pode auxiliar no controle de infecções por microrganismos patogênicos. / Antimicrobial host defense peptides are components of the innate immune system. They are expressed constitutively or induced by bacterial components, cytokines or the combination of both are expressed constitutively or induced by bacterial components, cytokines, or a combination of both. These peptides have direct action on microorganisms and are potent immunomodulators in mammalian cells. Microplusin is a copper II-chelating antimicrobial peptide from the Tick Rhipicephalus ( Boophilus) microplus, with a molecular mass of 10 204 Da. This peptide has a bacteriostatic activity against Micrococcus luteus, but no activity against E. coli and Candida albicans. The aim of the present work was to analyze the direct effect of microplusin in the culture of murine bone marrow derivated macrophages (BDM) treated or not with LPS or IFN-&#947. Our data showed that microplusin had no cytotoxic effect on macrophages, did not stimulate the synthesis of NO and IL-1&#946, but increased the production of TNF-&#945 and IL-6 after stimulation. The pre-treatment of macrophages with microplusin before LPS addition did not change the level of nitrite, TNF-&#945 or IL-6 in response to LPS. If the microplusin pre-treated macrophages were incubated with IFN-&#947 there was evidence that microplusin led to significant increase of NO and TNF-&#945 production confirming the pro-inflammatory characteristic of microplusin and its potential against pathogenic microorganisms. This indicates microplusin to be a potential immunomodulating agent of innate immunity and may help to control infections by pathogenic microorganisms.