Rheumatoid arthritis (RA) is a chronic joint disease which mechanism of action is unknown. The immune mechanism initiates an inflammatory process that mainly affects the joints, which can lead to cartilage and bone damage but also to some generalized symptoms such as fatigue and inflammatory anemia. At present, RA is primarily treated with methotrexate (MTX) as monotherapy. Studies in the field indicate that MTX in combination with infliximab, a monoclonal antibody, is more effective in treating RA than MTX treatment alone. Most likely, infliximab is not used as first-hand therapy since the reference product Remicade® is very expensive. In 2015 the patent for Remicade® expired and there are now cheaper biosimilars available that have been approved for the same indication as Remicade®. The aim of this study was to investigate whether MTX in combination with infliximab is more effective than MTX alone in the treatment of active RA and evaluating whether there is equivalence between the reference product Remicade® and the CT-P13 biosimilar. The medical and bioscience database PubMed was used to obtain the scientific articles analyzed in this literature study. From PubMed 5 articles were selected for further analysis. The delimitation of the search was: indication RA, MTX, infliximab and biosimilar (CT-P13). Articles 1-3, based on the outcome variables of the studies indicated that infliximab in combination with MTX is more effective than MTX treatment alone in the treatment of active RA. Articles 4-5 examined equivalence between the reference product for infliximab (RP) and the biosimilar CT-P13 in the treatment of active RA. The analysis and results of articles 4-5 showed that there was a similarity between RP and CT-P13 in treatment of active RA. Based on this small-scale literature study, treatment with MTX in combination with infliximab seems more beneficial for disease control in active RA than MTX treatment alone. In addition, the biosimilar CT-P13 appears to be equivalent to the reference product Remicade® in the treatment of active RA. Replacement of Remicade® to CT-P13 can mean major savings in healthcare and the therapeutic recommendation for active RA may need to be reviewed as there are now effective, cheaper alternatives.

Publicación a texto completo no autorizada por el autor / El documento digital no refiere asesor / Evalua la respuesta clínica al tratamiento con infliximab en pacientes con Artritis Reumatoide en el servicio de Reumatología del HNERM. Estudio de diseño descriptivo, realizado en el Servicio de Reumatología del Hospital Rebagliati, de los 35 pacientes registrados, se evaluó a 28. El promedio de edad fue de 52 años con una media del tiempo de duración de enfermedad de 14.46 años. El uso previo de metotrexate alcanzó el 100% y de leflunomida un 53.8%, ya sea como monoterapia o en terapia combinada. Se confirmó la disminución de los parámetros clínicos de actividad según el DAS28VSG desde 5.88 +-1.13 a 4.63+- 1.57 con un p= 0.0001 y CDAI de 29.4 +-12.04 a 18.61+-12.61 con un p=0.0001. La respuesta según los criterios EULAR fue buena en 21.4%, moderada en 35.7% e insatisfactoria en 42.8%. En el 25% de pacientes se suspendió el uso de infliximab por reacciones de hipersensibilidad. La respuesta clínica favorable medidos por DAS28VSG y respuesta EULAR fue menor a la esperada en los pacientes con artritis reumatoide. Se encontró alto porcentaje de pacientes que suspendieron el tratamiento por reacciones de hipersensibilidad. / Trabajo de investigación

Biologiska läkemedel är stora molekyler som har renats eller producerats från ett biologiskt ursprung. De används idag i stor utsträckning runt om i världen på grund av deras effektivitet att minska symtomen som kan uppstå vid olika mycket svåra sjukdomar. Detta arbete fokuserar på biologiska läkemedel som hämmar cytokinet tumörnekrotisk faktor (TNF). Läkemedlen används bland annat vid svåra autoimmuna sjukdomar. Vid autoimmuna sjukdomar aktiveras cellerna i immunsystemet vilket leder till utsöndring av olika cytokiner till exempel interleukin 1 och TNF. Dessa cytokiner påverkar kroppen på olika sätt genom att aktivera eller inhibera vissa produktionsprocesser i kroppen. Syftet med projektet var att verifiera en cellbaserad metod som används för koncentrationsmätning av det biologiska läkemedlet Infliximab (en TNF antagonist) i patientserum. Metoden som används i detta projekt kallas för reporter gene assay och den baseras på användning av en iLite cellinje. iLite är humana erytroleukemiceller K562 som är transfekterade med nukleär faktor kappa-B. Denna faktor aktiveras när TNF binder sig till TNF receptorer på cellytan. Aktiveringen av denna faktor leder till produktion av enzymet Eldflugeluciferas. Produktionen av enzymet är indirekt proportionell mot koncentrationen av TNF antagonist i serum. Dessutom innehåller cellerna en reporter gen för Renilla luciferas som uttrycks konstant. Renilla luciferas används som en internkontroll för att normalisera variationer som kan orsakas av provhantering samt transfektionseffektivitet. Luminescensen som utvecklas på grund av inverkan av Eldflugeluciferas på luciferas substrat samt inverkan av Renilla luciferas på stopp substrat mäts med hjälp av en luminometer. Dag-till-dag variationen för fem analyserade prov som analyserades vid fyra olika tillfällen var mellan 5 - 28 %, lot-till-lot variationen för fyra prov som analyserades fyra gånger vid fyra olika tillfällen var mellan 14 - 21 %. Alla normala serumprover från friska frivilliga blodgivare var negativa med en koncentration av Infliximab mindre än 0,65 µg/ ml. Resultaten visar att metoden är stabil och känslig jämfört med Biomonitors resultat. Dessutom visar resultaten att medelvärden av de olika körningarna från Euro Diagnostica (ett laboratorium som tillverkar immunologiska kit, Malmö) korrelerar väl med resultaten från Biomonitors laboratorium i Köpenhamn. Korrelationen för 19 analyserade prov var 0,99. / Biological drugs are large molecules that have been purified from or produced from a biological origin. They are used in great extent around the whole world because of their efficiency in reducing the symptoms of very severe diseases. The present work focus on biological drugs that inhibit tumor necrosis factor (TNF). Biological drugs are used for treatment of different autoimmune diseases. In autoimmune diseases activated cells of the immune system leads to secretion of various cytokines such as interleukin 1 and TNF. These cytokines affect the body in different ways by activating or inhibiting certain processes in the body. The purpose of this project was to verify a cell-based method which used for concentration measurement of the biological drug Infliximab (a TNF antagonist) in patient serum. The method which used in this project is called the reporter gene assay, and it is based on the use of an iLite cell line. iLite are human erythroleukemia K562 cells which are transfected with the nuclear factor kappa-B. This factor is activated when the tumor necrosis factor binds to its receptors on the cell surface. The activation of this factor leads to the production of the enzyme Firefly luciferase. Production of the enzyme is indirectly proportional to the concentration of TNF antagonist in the serum. Moreover, the cells contain a reporter gene for Renilla luciferase which is under constant expression. Renilla luciferase is used as an internal control to normalize the variations that may be caused by sample handling and transfection efficiency. The luminescence developed due to the influence of Firefly luciferase on luciferase substrate and Renilla luciferase on stop substrate is measured by using a luminometer. Day-to-day variations for five samples were analyzed at four different times were between 5 - 28 % while variations of lot-to-lot of 4 samples were analyzed at four different times were 14 - 21 %. The concentration of Infliximab in all the serum samples from healthy volunteer blood donors were less than 0.65 µg / ml. The results show that the method is robust and sensitive as compared to Biomonitors results. Furthermore, the results show that the mean value of the different runs from Euro Diagnostica (a laboratory that produces immunological kits, Malmö) for day-to-day and lot-to-lot variation agrees quite well with those from Biomonitors laboratory in Copenhagen. The correlation for the nineteen analyzed samples was 0.99.

IN ENGLISH Drug dependency in inflammatory bowel disease (IBD), Crohn's disease (CD) and ulcerative colitis (UC), is a specific disease phenotype which determines disease prognosis and hence may be used as a prognostic marker for treatment management. Drug dependency in IBD has been well described in corticosteroid treatment and recently also in infliximab (IFX) therapy. The aims of this thesis were: 1) to assess the occurrence of IFX dependency in paediatric and adult patients with CD; further to search for clinical and genetic predictors of IFX outcome and to evaluate the impact of IFX dependency on surgical rate; 2) to assess in CD patients the outcome of the first course of 5-ASA monotherapy with emphasis on 5-ASA dependency and to define clinical predictors of 5-ASA treatment outcome. We found that 66% of children and 29% of adults with CD became IFX dependent. The high frequency in paediatrics is in agreement with previously published studies, while the finding in adult patients indicates a lower rate of IFX dependency in the only study to date. Perianal disease and no bowel surgery prior to IFX start were predicative of IFX dependency in paediatric patients. In adult cohort, 2 genetic variants LTA c.207 A>G and CASP9 c.93 C>T were associated with IFX outcome, whereas no relevant clinical...

L’utilisation d’agents anti-TNF a grandement amélioré la prise en charge de certaines maladies inflammatoires chroniques comme la polyarthrite rhumatoïde (PR) ou les maladies inflammatoires chroniques de l’intestin (MICI). Cependant, le quart des patients environ ne vont pas répondre au traitement ou présenteront une perte de réponse secondaire. Des marqueurs prédictifs de réponse sont nécessaires afin limiter les effets secondaires et les coûts inutiles en ciblant les patients qui pourraient être améliorés par les anti-TNF. Ces travaux de recherche se sont dans un premier temps concentrés sur l’importance de l’immunogénicité de ces traitements. Des anticorps anti-médicaments (ADAs) étaient bien associés à un taux bas d’anti-TNF avec des conséquences cliniques en termes de perte de réponse clinique et d’absence de cicatrisation muqueuse dans les MICI. Des seuils cliniques d’interprétation des tests biologiques pour la détection du médicament et de ses anticorps ont pu être définis et correspondent à 4.9 μg/ml pour l’infliximab et 200 ng/ml pour les ADAs. Ces résultats obtenus par ELISA sont bien corrélés avec les tests fonctionnels réalisés en parallèle et confirment l’intérêt de cette technique dans ce dépistage. Les ADAs étaient diminués par traitement immunosuppresseur concomitant. Ensuite, la persistance d’une infection chronique mise en évidence par des anticorps anti-bactériens a été évaluée en tant que marqueur prédictif de réponse aux anti-TNF. Aucun résultat statistiquement significatif n’a pu être relevé sur ces premières données, que ça soit pour les anticorps dirigés contre la flore intestinale pour les MICI ou contre le microbiote oral dans la PR. Seul un taux élevé de MMP-3 à l’initiation de l’infliximab chez les patients PR prédisait d’une bonne réponse clinique selon les critères de l’EULAR par la suite / The use of anti-TNF agents has greatly improved the management of chronic inflammatory diseases such as rheumatoid arthritis (RA), or chronic inflammatory bowel disease (IBD). However, about a quarter of patients will not respond to treatment or will present a secondary loss of response. Predictive biomarkers of response are needed to reduce side effects and unnecessary costs by targeting patients that could be improved by anti-TNF. This research work was initially focused on the importance of immunogenicity of these treatments. Anti-drug antibodies (ADAs) were well associated with low levels of anti-TNF with clinical consequences in terms of loss of clinical response and absence of mucosal healing in IBD. Clinical thresholds for drug and ADAs have been defined and correspond to 4.9 μg/ml for infliximab and 200 ng/ml for ADAS. These results obtained by ELISA correlate well with functional tests done in parallel, and confirm the value of this technique for screening. The ADAs were decreased with concomitant immunosuppressive therapy. Then, the persistence of chronic infection as evidenced by anti-bacterial antibody was evaluated as a predictive marker for response to anti-TNF. No statistically significant results could be raised on these first data, for antibodies against the intestinal flora in IBD or against the oral microbiota in RA. Only high levels of MMP-3 at the initiation of infliximab in RA patients predicted a good clinical response according to the EULAR criteria

IN ENGLISH Drug dependency in inflammatory bowel disease (IBD), Crohn's disease (CD) and ulcerative colitis (UC), is a specific disease phenotype which determines disease prognosis and hence may be used as a prognostic marker for treatment management. Drug dependency in IBD has been well described in corticosteroid treatment and recently also in infliximab (IFX) therapy. The aims of this thesis were: 1) to assess the occurrence of IFX dependency in paediatric and adult patients with CD; further to search for clinical and genetic predictors of IFX outcome and to evaluate the impact of IFX dependency on surgical rate; 2) to assess in CD patients the outcome of the first course of 5-ASA monotherapy with emphasis on 5-ASA dependency and to define clinical predictors of 5-ASA treatment outcome. We found that 66% of children and 29% of adults with CD became IFX dependent. The high frequency in paediatrics is in agreement with previously published studies, while the finding in adult patients indicates a lower rate of IFX dependency in the only study to date. Perianal disease and no bowel surgery prior to IFX start were predicative of IFX dependency in paediatric patients. In adult cohort, 2 genetic variants LTA c.207 A>G and CASP9 c.93 C>T were associated with IFX outcome, whereas no relevant clinical...

Before market authorization of new medicines, their efficacy and safety are evaluated using randomized controlled trials. While there is no doubt about the scientific value of randomized trials, they are usually conducted in selected populations with questionable generalizability to routine care.  In the digital data revolution era, with healthcare data growing at an unprecedented rate, drug monitoring in routine care is still highly under-utilized. Although many countries have access to data on prescription drugs at the individual level in ambulatory care, such data are often missing for hospitals. This is a growing problem considering the clear trend towards more new and expensive drugs administered in the hospital setting. The aim of this thesis was therefore to develop methods for extracting data on drug use from a hospital-based electronic health record system and further to build and evaluate models for real-time monitoring of effectiveness and safety of new drugs in routine care using data from electronic health records and regional and national health care registers. Using the developed techniques, we were able to demonstrate drug use and health service utilization for inflammatory bowel disease and to evaluate the comparative effectiveness and safety of antiarrhythmic drugs. With a rapidly evolving drug development, it is important to optimize the evaluation of effectiveness, safety and health economic value of new medicines in routine care. We believe that the models described in this thesis could contribute to fulfil this need.

O tratamento de pacientes apresentando doenças inflamatórias imunomediadas com drogas anti-TNF-alfa aumenta o risco da reativação da tuberculose. Isso sugere que tais drogas possam afetar a imunidade celular destes. No entanto, há dados conflitantes sobre se esse tratamento suprime as respostas para o teste tuberculínico (TT) e os ensaios de liberação de interferon-gama (IGRAs) e poucos dados em pacientes com psoríase. O presente estudo avaliou pacientes com psoríase moderada a grave enfocando os efeitos do tratamento com infliximabe em suas respostas imunológicas celulares. Foram avaliadas as respostas imunes celulares de doze pacientes antes e durante o tratamento com infliximabe. As células mononucleares do sangue periférico (PBMC) foram estimuladas com a fito-hemaglutinina (PHA), o superantígeno enterotoxina B (SEB), um lisado de citomegalovírus (CMV), e antígenos de Mycobacterium tuberculosis, e a ativação de linfócitos foi avaliada por ELISPOT para enumerar células secretoras de IFN-y, por ELISA para detecção da secreção de IFN-y, e através da incorporação de[3H] timidina para medir a proliferação. O tratamento com infliximabe não levou à redução de INF-y e da resposta linfoproliferativa nos pacientes. Pelo contrário, aumentou a liberação desta citocina em culturas de PBMC estimulados com PHA e SEB por 12 h. Este efeito foi mais notado no pico do efeito clínico do agente anti-TNF (7 semanas de tratamento) e menos proeminente no seu nadir (logo antes da infusão da próxima dose). Reatividade imunitária ao CMV também não foi significativamente afetada, notando-se leve aumento pelo agente anti-TNF. É de se notar que secreção de IFN-y e resposta proliferativa a Mtb dos dois pacientes TT positivos foram, também, visivelmente aumentadas na semana 7, declinando quando infliximabe atingiu o seu nadir. Os efeitos deletérios do bloqueio do TNF em pacientes com psoríase grave, submetidos ao tratamento com infliximabe parecem ser atenuados, pelo menos parcialmente, por um efeito de aumento na imunidade mediada por células desses pacientes, possivelmente pela diminuição da ação imunossupressora decorrente do excesso de TNF / Treatment of patients with immune-mediated inflammatory diseases with anti-TNF agents increases the risk of tuberculosis reactivation, suggesting that it may affect their cellular immune response. However, there are conflicting data on whether anti-TNF treatment suppresses the responses to tuberculin skin test (TST) and interferon-y release assays and no information regarding psoriasis patients on anti-TNF treatment. The present study evaluated patients with moderate to severe psoriasis focusing on the effects of treatment with infliximab on their cellular immune responses. Cellular immune responses of twelve patients were evaluated before and during infliximab treatment. Peripheral blood mononuclear cells (PBMC) were stimulated with phytohemaglutinin (PHA), the superantigen enterotoxin B (SEB), a cytomegalovirus lysate (CMV), and Mycobacterium tuberculosis antigens, and the lymphocyte activation was evaluated by ELISPOT for enumeration of IFN-y-secreting cells, ELISA for detection of secreted IFN-y, and by [3H]thymidine incorporation for proliferation measurement. Treatment with infliximab does not lead to reduction in the INF-y and lymphoproliferative responses of patients. It rather increased the overnight release of this cytokine in PBMC cultures stimulated with PHA and SEB. This effect was most noted at the peak of the clinical effect of the anti-TNF agent (week 7 of treatment) and less prominent at its nadir (just before infusion of the next dose). Immune reactivity to CMV was also either unaffected or slightly increased by the TNF blocking agent. Of note, the IFN-y and proliferative responses to Mtb from the two TST-responder patients were also remarkably increased at week 7, declining when infliximab reached its nadir. The deleterious consequences of TNF blockade in patients with severe psoriasis undergoing infliximab treatment may be in part attenuated by an enhancing effect on the cell mediated immunity of the patients, possibly due to the abbreviation of the immunosuppressive effect of TNF overexpression

O tratamento de pacientes apresentando doenças inflamatórias imunomediadas com drogas anti-TNF-alfa aumenta o risco da reativação da tuberculose. Isso sugere que tais drogas possam afetar a imunidade celular destes. No entanto, há dados conflitantes sobre se esse tratamento suprime as respostas para o teste tuberculínico (TT) e os ensaios de liberação de interferon-gama (IGRAs) e poucos dados em pacientes com psoríase. O presente estudo avaliou pacientes com psoríase moderada a grave enfocando os efeitos do tratamento com infliximabe em suas respostas imunológicas celulares. Foram avaliadas as respostas imunes celulares de doze pacientes antes e durante o tratamento com infliximabe. As células mononucleares do sangue periférico (PBMC) foram estimuladas com a fito-hemaglutinina (PHA), o superantígeno enterotoxina B (SEB), um lisado de citomegalovírus (CMV), e antígenos de Mycobacterium tuberculosis, e a ativação de linfócitos foi avaliada por ELISPOT para enumerar células secretoras de IFN-y, por ELISA para detecção da secreção de IFN-y, e através da incorporação de[3H] timidina para medir a proliferação. O tratamento com infliximabe não levou à redução de INF-y e da resposta linfoproliferativa nos pacientes. Pelo contrário, aumentou a liberação desta citocina em culturas de PBMC estimulados com PHA e SEB por 12 h. Este efeito foi mais notado no pico do efeito clínico do agente anti-TNF (7 semanas de tratamento) e menos proeminente no seu nadir (logo antes da infusão da próxima dose). Reatividade imunitária ao CMV também não foi significativamente afetada, notando-se leve aumento pelo agente anti-TNF. É de se notar que secreção de IFN-y e resposta proliferativa a Mtb dos dois pacientes TT positivos foram, também, visivelmente aumentadas na semana 7, declinando quando infliximabe atingiu o seu nadir. Os efeitos deletérios do bloqueio do TNF em pacientes com psoríase grave, submetidos ao tratamento com infliximabe parecem ser atenuados, pelo menos parcialmente, por um efeito de aumento na imunidade mediada por células desses pacientes, possivelmente pela diminuição da ação imunossupressora decorrente do excesso de TNF / Treatment of patients with immune-mediated inflammatory diseases with anti-TNF agents increases the risk of tuberculosis reactivation, suggesting that it may affect their cellular immune response. However, there are conflicting data on whether anti-TNF treatment suppresses the responses to tuberculin skin test (TST) and interferon-y release assays and no information regarding psoriasis patients on anti-TNF treatment. The present study evaluated patients with moderate to severe psoriasis focusing on the effects of treatment with infliximab on their cellular immune responses. Cellular immune responses of twelve patients were evaluated before and during infliximab treatment. Peripheral blood mononuclear cells (PBMC) were stimulated with phytohemaglutinin (PHA), the superantigen enterotoxin B (SEB), a cytomegalovirus lysate (CMV), and Mycobacterium tuberculosis antigens, and the lymphocyte activation was evaluated by ELISPOT for enumeration of IFN-y-secreting cells, ELISA for detection of secreted IFN-y, and by [3H]thymidine incorporation for proliferation measurement. Treatment with infliximab does not lead to reduction in the INF-y and lymphoproliferative responses of patients. It rather increased the overnight release of this cytokine in PBMC cultures stimulated with PHA and SEB. This effect was most noted at the peak of the clinical effect of the anti-TNF agent (week 7 of treatment) and less prominent at its nadir (just before infusion of the next dose). Immune reactivity to CMV was also either unaffected or slightly increased by the TNF blocking agent. Of note, the IFN-y and proliferative responses to Mtb from the two TST-responder patients were also remarkably increased at week 7, declining when infliximab reached its nadir. The deleterious consequences of TNF blockade in patients with severe psoriasis undergoing infliximab treatment may be in part attenuated by an enhancing effect on the cell mediated immunity of the patients, possibly due to the abbreviation of the immunosuppressive effect of TNF overexpression

La rectocolite hémorragique (RCH) est une maladie inflammatoire chronique de l'intestin responsable d'un handicap et d'une altération de la qualité de vie pouvant exposer les patients à des complications sévères en dépit des thérapeutiques actuelles. L'objectif de cette thèse était d'analyser les voies possibles d'amélioration de la prise en charge thérapeutique des patients à partir de données expérimentales et cliniques. Au niveau expérimentale nous nous sommes intéressés au mécanisme impliqué dans la régulation de l'expression de PPARγ, récepteur nucléaires aux propriétés anti-inflammatoires, primitivement diminuée au cours de la RCH et cible des 5-aminosalicylés. Il a été montré que son expression était d'une part sous le contrôle de la stéroidogenèse intraépithéliale, elle-même sous contrôlée par LRH-1 et d'autre part que l'hypoxie épithéliale diminuait son expression via une sur-expression de miR-27a. Par ailleurs, les effets de l'hypoxie sur l'expression de PPARγ étaient inversés en présence de sildénafil. Sur le versant clinique, l'analyse d'une nouvelle stratégie thérapeutique ciblant la cicatrisation muqueuse, c'est à dire l'abrogation de l'inflammation colique macroscopique était efficace et possible dans la pratique clinique. Cet objectif semble aujourd'hui fondamental pour diminué la morbi-mortalité induite par cette maladie. Enfin nous avons observé l'efficacité des anti-TNF dans le cas spécifique de la rectite réfractaire et l'importance de son utilisation prolongée pour éviter les rechutes de la maladie et l'obtention d'une rémission prolongée.