La nature du lien qui unit les deux aspects du système immunitaire, que sont la défense de l’hôte contre les agressions extérieures et la genèse des maladies auto- immunes, n’a pas été élucidée. L’axe IL-23/Th17 joue un rôle dans les deux versants, ce qui le place en bonne position pour être un potentiel chaînon manquant. Objectif : connaître l’implication de cet axe dans un modèle infectieux, Mycoplasma hominis, et un modèle de maladie auto-immune, la sclérodermie systémique (ScS), dans lesquels il n’a pas encore été étudié. Les lipoprotéines membranaires de M. hominis sont capables d’induire une maturation des cellules dendritiques humaines. Elle s’accompagne d’une sécrétion d’IL-23 variable selon l’origine clinique des isolats, via TLR2, et d’une polarisation vers la voie Th17. Nous avons observé une augmentation de la fréquence des cellules Th17 et Th22 dans le sang périphérique des patients ScS, potentialisée par l’iloprost, via entre autres la production monocytaire d’IL-23. Dans la peau des patients ScS, il existe une augmentation des cellules produisant l’IL-17 inversement corrélé au score de fibrose cutanée. In vitro, l’IL-17 est capable d’inhiber partiellement l’expression d’α-SMA induite par le TGF-ß et d’induire la sécrétion de MMP1 par des fibroblastes dermiques humains. L’axe IL-23/IL-17 et les cellules Th17 jouent un rôle dans la défense contre M. hominis et dans la physiopathologie de la ScS. / Relationship between both aspects of the immune system, ie host defense against external aggression and genesis of autoimmune diseases, has not been elucidated. IL-23/Th17 axis plays a role in both sides, which puts him in a good position to be a potential missing link. Objective: To understand the implication of this axis in a model of infection, Mycoplasma hominis, and a model of autoimmune disease, systemic sclerosis (SSc), in which it has not yet been studied.
The membrane lipoproteins of M. hominis are capable of inducing human dendritic cell maturation. It occurs along with an IL-23 secretion changing with the clinical origin of isolates, via TLR2, and a T cell polarization towards Th17. Then we observed an increase in the Th17 and Th22 cell frequency in peripheral blood of SSc patients, further enhanced by iloprost via monocyte production of IL-23 among others. In the skin of SSc patients, we showed an increase in IL-17-producing cells with an inverse correlation to the skin fibrosis score. In vitro, IL-17 is able to partially inhibit the expression of α-SMA induced by TGF-ß and to induce the secretion of MMP1 in human dermal fibroblasts. The IL-23/IL-17 axis and Th17 cells play a role in defense against M. hominis and in the pathogenesis of SSc.

A leishmaniose tegumentar americana (LTA) apresenta um amplo espectro de manifestações clínicas e imunopatológicas resultante da interação entre as diferentes espécies de Leishmania e os mecanismos de resposta imune do hospedeiro. Leishmania (Viannia) braziliensis e Leishmania (Leishmania) amazonensis são as espécies de maior potencial patogênico para o homem e de importância médica no Brasil. As células TCD4, quando ativadas por antígenos via MHC II podem se diferenciar em linhagens de células efetoras como Th1, Th2, Th17 e células T reguladoras (Treg). IL-23 é indispensável para as funções efetoras e manutenção de células Th17. O objetivo deste trabalho foi avaliar a expressão de Foxp3, IL-17 e IL-23 em lesões cutâneas de pacientes com diferentes formas clínicas da LTA. Biópsias parafinadas de 44 pacientes foram submetidas à imunoistoquímica, sendo 6 casos de leishmaniose cutânea anérgica difusa (LCADIDRM-) e leishmaniose cutânea disseminada borderline (LCDBIDRM-), ambas causadas por L.(L) amazonensis e 16 casos de leishmaniose cutânea localizada (LCLIDRM+) também causada por L.(L.) amazonensis; 9 casos de LCLIDRM+, 2 casos de LCDBIDRM- e 5 casos de leishmaniose cutâneo-mucosa (LCMIDRM+), todos causados por L.(V.) braziliensis. A densidade de células Tregs Foxp3+ no espectro clínico da LTA mostrou um aumento progressivo partindo das formas centrais LCL causadas por L.(V.) braziliensis (170mm2) e L.(L) amazonensis (140mm2) para as formas polares, LCADIDRM- (289mm2) e LCDBIDRM- (183mm2) causada por L.(L) amazonensis, LCDBIDRM- (189mm2) e LCMIDRM+, causadas por L.(V.) braziliensis (158mm2). A comparação entre as densidades de células IL-17+ nas diferentes formas clínicas da LTA mostrou um perfil semelhante também com um aumento progressivo da expressão de IL-17 partindo das formas centrais LCLIDRM+ causadas por L.(V.) braziliensis (232mm2) e L.(L) amazonensis (197mm2) em direção as formas polares, LCADIDRM- (470mm2) e LCDBIDRM- (340mm2) causada por L.(L.) amazonensis, LCDBIDRM- (431mm2) e LCMIDRM+ (372mm2) causada por L.(V.) braziliensis. A densidade de células IL-23+ mostrou perfil similar ao de IL-17 como no espectro de doença causada por L. (V.) braziliensis ou L. (L.) amazonensis: LCADIDRM- (687mm2), LCDBIDRM- (518mm2) e LCLIDRM+ (348mm2) por L.(L.) amazonensis, LCLIDRM+ (457mm2), LCDBIDRM- (609mm2) e LCMIDRM+ (568mm2) L. (V.) braziliensis. Diante dos nossos achados, observa-se que as células Foxp3+, IL-17+ e IL-23+ desempenham um papel importante na imunopatogênese das diferentes formas clínicas da LTA causadas por L. (V.) braziliensis ou L. (L.) amazonensis, caracterizada por uma resposta imune polarizada de diferente expressão patológica / The American cutaneous leishmaniasis (ACL) presents a wide spectrum of clinical and immunopathological manifestations resulting from the interaction between the different species of Leishmania and the mechanisms of the host immune response. Leishmania (Viannia) braziliensis and Leishmania (Leishmania) amazonensis are the species with the largest pathogenic potential for humans and medical importance in Brazil. The CD4+ T cells can be differentiated into effector cell lines as Th1, Th2, Th17 and regulatory T cells (Treg). IL-23 is essential for effector functions and maintenance of Th17 cells, that produces IL-17. The aim of this study was to evaluate the expression of Foxp3, IL-17 and IL-23 in cutaneous lesions of patients with different clinical forms of ACL. Paraffin embedded biopsies from 44 patients were submitted to immunohistochemistry, there were 6 cases of anergic diffuse cutaneous leishmaniasis (ADCLDTH-) and borderline disseminated cutaneous leishmaniasis (BDCLDTH-) both caused by L. (L.) amazonensis 16 cases of cutaneous leishmaniasis (LCLDTH+) caused by L. (L.) amazonensis, 9 cases of LCLDTH+, 2 cases of BDCLDTH- and 5 cases of mucocutaneous leishmaniasis (MCLDTH+) all caused by L. (V.) braziliensis. The density of Treg Foxp3+ cells in the clinical spectrum of ACL showed a progressive increase starting from the central forms LCLDTH+ caused by L. (V.) braziliensis (170mm2) and L. (L) amazonensis (140mm2) towards the polar forms ADCLDTH- (289mm2). The intermediate clinical forms BDCLDTH- (183mm2) caused by L. (L) amazonensis and BDCLDTH-(189mm2) by L. (V.) braziliensis as well as, MCLDTH+(158mm2) did not present any significant differences. The comparison between the densities of IL-17+ cells in different clinical forms of ACL showed progressive increasing starting from the central forms LCLDTH+ caused by L. (V.) braziliensis (232mm2) and L. (L) amazonensis (197mm2) towards the polar forms, ADCLDTH-(470mm2) and BDCLDTH-(340mm2) caused by L. (L.) amazonensis BDCLDTH- (431mm2) and MCLDTH+ (372mm2) caused by L. (V.) braziliensis. The density of IL-23+ cells showed a similar profile to that of IL-17 at the disease spectrum caused by L. (V.) braziliensis and L. (L.) amazonensis: ADCLDTH-(687mm2) BDCLDTH-(518mm2) and LCLDTH+ (348mm2) by L. (L.) amazonensis; LCLDTH+ (457mm2) LCDBDTH-(609mm2) and MCLDTH+ (568mm2) L. (V.) braziliensis. In view of our findings, we notice that the Foxp3+, IL-17+ and IL-23+ cells play an important role in the immunopathogenesis of different clinical forms of ACL caused by L. (V.) braziliensis and L. (L.) amazonensis, characterized by an immune polarized response with different pathological expression

Influenza virus is one of the major respiratory pathogens of humans as well as animals, including equines. There is an increasing evidence that bacterial infections are the most common cause of the death during influenza. In horses also, secondary bacterial pneumonia can lead to death, and surviving horses may take up to six months for the complete recovery resulting in heavy economic loss to the equine industry. Interleukin (IL)-23 mediated innate immune response has been shown to protect the host from various respiratory bacterial infections. However, studies to investigate the role of host and viral factors in the regulation of IL-23 are limited. Endoplasmic reticulum (ER) stress-induced transcription factor CHOP-10 and IFN-β has been shown to participate in the regulation of IL-23. Primary hypothesis for the current study was that influenza A virus (IAV) NS1 protein downregulates the IL-23 expression via inhibition of CHOP-10. In order to test our hypothesis, we infected the RAW264.7 cells - a murine macrophage cell line, and primary murine alveolar macrophage cells either with the wild type Influenza A virus (PR/8/34, PR8) or isogenic mutant virus lacking NS1 (delNS1). Quantitative analysis of mRNA expression revealed a significantly higher mRNA expression of IL23p19, IFN-β and CHOP-10 in delNS1 virus infected cells as compared the PR8 virus infected cells. Additionally, overexpression of CHOP-10 partially restored the expression of IL-23p19 in PR8 virus infected cells and knockdown of CHOP-10 resulted in downregulated expression of IL-23p19 in delNS1 infected cells. Taken together, these results suggest that IAV NS1 protein mediated inhibition of CHOP-10 expression leads to downregulation of IL-23 expression in macrophage cells in-vitro. Similar results were also observed in-vivo using IAV and Streptococcus zoooepidemicus (S. ze) co-infection model. In a co-infection mouse model delNS1 virus co-infection resulted in significantly higher expression of the IL-23 and IL-17. Considering the role of IL-23 in protection against respiratory bacterial pathogens, effect of exogenous supplementation of IL-23 was also investigated in the influenza and S. ze co-infection mouse model. We found that a single intranasal dose of recombinant murine IL-23 significantly improved the survival of mice co-infected with PR8 and S .ze. Overall, our study suggests that IAV infection subverts the IL-23 mediated respiratory innate immune response and restoration of IL-23 could protect from influenza-associated respiratory bacterial infections.

Made available in DSpace on 2014-12-17T15:32:21Z (GMT). No. of bitstreams: 1 BarbaraVBM_DISSERT.pdf: 1563576 bytes, checksum: c536e73cdfce0251b3446e74e0f9cc69 (MD5) Previous issue date: 2012-02-24 / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico / Th17 cells have been strongly associated with the pathogenesis of several autoimmune and inflammatory diseases. IL-17 and IL-23 are important cytokines associated with this lineage. The aim of this study was to analyze, through immunohistochemical methods, the immunoexpression of IL-17 and IL-23 in the inflammatory infiltrate of oral lichen planus (OLP) lesion compared to that of inflammatory fibrous hyperplasia (IFH) and between clinical forms reticular and erosive of OLP. The sample included 41 cases of OLP, of which 23 were reticular and 18 erosive and 10 cases of IFH. The results were subjected to nonparametric statistical tests with a 5% significance level. In OLP lesions histomorphological analysis, the most common findings were: hyperparakeratinization, specimens with atrophic epithelium in erosive clinical form (p = 0.011), epithelial projections in most of reticular type of lesions, in addition Civatte bodies were identified in most samples of both clinical forms. For immunohistochemistry analysis, five fields with strong immunoreactivity for IL-17 and IL-23 were photomicrographed at 400x magnification, images were transferred to a computer where with ImageJ software?, lymphocytes that exhibited cytoplasmic immunostaining for these cytokines were counted. A mean was established after for each case. There was no statistically significant difference in the number of imunopositive lymphocytes for IL-17 and IL-23 among the group of OLP and IFH group, however a larger amount of lymphocytes imunopositive for IL-17 was found in the LPO group (p = 0.079) and significantly higher amounts of those lymphocytes were found in the erosive OLP when compared to the group of reticular OLP and IFH (p = 0.019). Furthermore, a marker epithelial immunopositivity for IL-17 was observed in OLP group. Although the results of this study do not permit the forceful assertion about the participation of Th17 lineage in OLP lesions, the findings of immunopositive lymphocytes counting for IL-17 and IL-23, which are potent proinflammatory cytokines, together with the the marked epithelial immunopositivity found for IL-17 in this study, suggest a possible role of this lineage in the pathogenesis of this disorder / As c?lulas Th17 t?m sido fortemente associadas com a patogenia de diversas doen?as autoimunes e inflamat?rias. A IL-17 e a IL-23 s?o importantes citocinas associadas com esta linhagem. O objetivo do presente trabalho foi analisar, atrav?s de m?todos imunohistoqu?micos, a imunoexpress?o da IL-17 e da IL-23 no infiltrado inflamat?rio das les?es de l?quen plano oral (LPO) comparando ao da hiperplasia fibrosa inflamat?ria (HFI) e entre as formas cl?nicas reticular e erosiva do LPO com o intuito de esclarecer se a linhagem Th17 participa da patog?nese do LPO. Na amostra foram inclu?dos 41 casos de LPO, dos quais 23 eram reticulares e 18 erosivos, al?m de 10 casos de HFI. Os resultados foram submetidos a testes estat?sticos n?o param?tricos com n?vel de signific?ncia de 5%. Na an?lise histomorfol?gica das les?es de LPO, observou-se predom?nio de: les?es hiperparaceratinizadas, esp?cimes com epit?lio atr?fico na forma cl?nica erosiva (p=0,011), proje??es epiteliais nas les?es do tipo reticular, al?m de corpos de Civatte identificados na maior parte da amostra de ambas as formas cl?nicas. Para o estudo imuno-histoqu?mico, cinco campos com forte imunorreatividade para a IL-17 e para a IL-23 foram fotomicrografados sob o aumento de 400x, as fotos foram transferidas para um computador onde com o aux?lio do software ImageJ?, realizou-se a contagem dos linf?citos que exibiram imunomarca??o citoplasm?tica para estas citocinas. Posteriormente, foi estabelecida uma m?dia para cada caso. N?o foram observadas diferen?as estatisticamente significativas na quantidade de linf?citos imunopositivos para a IL-17 e para a IL-23 entre o grupo do LPO e da HFI, no entanto uma maior quantidade desses linf?citos para a IL-17 foi encontrada no grupo do LPO (p=0,079) e uma quantidade significativamente maior de linf?citos imunopositivos para a IL- 23 foi encontrada entre o grupo do LPO erosivo e da HFI (p=0,019). Al?m disto, foi observada uma marcante imunopositividade epitelial para a IL-17 no grupo do LPO. Ainda que os resultados do presente estudo n?o permitam a afirma??o contundente da participa??o da linhagem Th17 nas les?es de LPO, os achados da contagem dos linf?citos imunopositivos para a IL-17 e para a IL-23, que s?o potentes citocinas pr?-inflamat?rias, somados ? marcante imunopositividade epitelial encontrada para a IL-17 neste estudo, sugerem uma poss?vel participa??o desta linhagem na patog?nese desta desordem

A leishmaniose tegumentar americana (LTA) apresenta um amplo espectro de manifestações clínicas e imunopatológicas resultante da interação entre as diferentes espécies de Leishmania e os mecanismos de resposta imune do hospedeiro. Leishmania (Viannia) braziliensis e Leishmania (Leishmania) amazonensis são as espécies de maior potencial patogênico para o homem e de importância médica no Brasil. As células TCD4, quando ativadas por antígenos via MHC II podem se diferenciar em linhagens de células efetoras como Th1, Th2, Th17 e células T reguladoras (Treg). IL-23 é indispensável para as funções efetoras e manutenção de células Th17. O objetivo deste trabalho foi avaliar a expressão de Foxp3, IL-17 e IL-23 em lesões cutâneas de pacientes com diferentes formas clínicas da LTA. Biópsias parafinadas de 44 pacientes foram submetidas à imunoistoquímica, sendo 6 casos de leishmaniose cutânea anérgica difusa (LCADIDRM-) e leishmaniose cutânea disseminada borderline (LCDBIDRM-), ambas causadas por L.(L) amazonensis e 16 casos de leishmaniose cutânea localizada (LCLIDRM+) também causada por L.(L.) amazonensis; 9 casos de LCLIDRM+, 2 casos de LCDBIDRM- e 5 casos de leishmaniose cutâneo-mucosa (LCMIDRM+), todos causados por L.(V.) braziliensis. A densidade de células Tregs Foxp3+ no espectro clínico da LTA mostrou um aumento progressivo partindo das formas centrais LCL causadas por L.(V.) braziliensis (170mm2) e L.(L) amazonensis (140mm2) para as formas polares, LCADIDRM- (289mm2) e LCDBIDRM- (183mm2) causada por L.(L) amazonensis, LCDBIDRM- (189mm2) e LCMIDRM+, causadas por L.(V.) braziliensis (158mm2). A comparação entre as densidades de células IL-17+ nas diferentes formas clínicas da LTA mostrou um perfil semelhante também com um aumento progressivo da expressão de IL-17 partindo das formas centrais LCLIDRM+ causadas por L.(V.) braziliensis (232mm2) e L.(L) amazonensis (197mm2) em direção as formas polares, LCADIDRM- (470mm2) e LCDBIDRM- (340mm2) causada por L.(L.) amazonensis, LCDBIDRM- (431mm2) e LCMIDRM+ (372mm2) causada por L.(V.) braziliensis. A densidade de células IL-23+ mostrou perfil similar ao de IL-17 como no espectro de doença causada por L. (V.) braziliensis ou L. (L.) amazonensis: LCADIDRM- (687mm2), LCDBIDRM- (518mm2) e LCLIDRM+ (348mm2) por L.(L.) amazonensis, LCLIDRM+ (457mm2), LCDBIDRM- (609mm2) e LCMIDRM+ (568mm2) L. (V.) braziliensis. Diante dos nossos achados, observa-se que as células Foxp3+, IL-17+ e IL-23+ desempenham um papel importante na imunopatogênese das diferentes formas clínicas da LTA causadas por L. (V.) braziliensis ou L. (L.) amazonensis, caracterizada por uma resposta imune polarizada de diferente expressão patológica / The American cutaneous leishmaniasis (ACL) presents a wide spectrum of clinical and immunopathological manifestations resulting from the interaction between the different species of Leishmania and the mechanisms of the host immune response. Leishmania (Viannia) braziliensis and Leishmania (Leishmania) amazonensis are the species with the largest pathogenic potential for humans and medical importance in Brazil. The CD4+ T cells can be differentiated into effector cell lines as Th1, Th2, Th17 and regulatory T cells (Treg). IL-23 is essential for effector functions and maintenance of Th17 cells, that produces IL-17. The aim of this study was to evaluate the expression of Foxp3, IL-17 and IL-23 in cutaneous lesions of patients with different clinical forms of ACL. Paraffin embedded biopsies from 44 patients were submitted to immunohistochemistry, there were 6 cases of anergic diffuse cutaneous leishmaniasis (ADCLDTH-) and borderline disseminated cutaneous leishmaniasis (BDCLDTH-) both caused by L. (L.) amazonensis 16 cases of cutaneous leishmaniasis (LCLDTH+) caused by L. (L.) amazonensis, 9 cases of LCLDTH+, 2 cases of BDCLDTH- and 5 cases of mucocutaneous leishmaniasis (MCLDTH+) all caused by L. (V.) braziliensis. The density of Treg Foxp3+ cells in the clinical spectrum of ACL showed a progressive increase starting from the central forms LCLDTH+ caused by L. (V.) braziliensis (170mm2) and L. (L) amazonensis (140mm2) towards the polar forms ADCLDTH- (289mm2). The intermediate clinical forms BDCLDTH- (183mm2) caused by L. (L) amazonensis and BDCLDTH-(189mm2) by L. (V.) braziliensis as well as, MCLDTH+(158mm2) did not present any significant differences. The comparison between the densities of IL-17+ cells in different clinical forms of ACL showed progressive increasing starting from the central forms LCLDTH+ caused by L. (V.) braziliensis (232mm2) and L. (L) amazonensis (197mm2) towards the polar forms, ADCLDTH-(470mm2) and BDCLDTH-(340mm2) caused by L. (L.) amazonensis BDCLDTH- (431mm2) and MCLDTH+ (372mm2) caused by L. (V.) braziliensis. The density of IL-23+ cells showed a similar profile to that of IL-17 at the disease spectrum caused by L. (V.) braziliensis and L. (L.) amazonensis: ADCLDTH-(687mm2) BDCLDTH-(518mm2) and LCLDTH+ (348mm2) by L. (L.) amazonensis; LCLDTH+ (457mm2) LCDBDTH-(609mm2) and MCLDTH+ (568mm2) L. (V.) braziliensis. In view of our findings, we notice that the Foxp3+, IL-17+ and IL-23+ cells play an important role in the immunopathogenesis of different clinical forms of ACL caused by L. (V.) braziliensis and L. (L.) amazonensis, characterized by an immune polarized response with different pathological expression

Aspergillus fumigatus is an opportunistic fungal pathogen that causes lethal invasive pulmonary disease in immunocompromised hosts and allergic asthma in sensitized individuals. This dissertation explores how eosinophils may protect hosts from acute infection while driving asthma pathogenesis by co-producing IL-23 and IL-17 in both contexts. In an acute model of pulmonary aspergillosis, eosinophils were observed to associate with and kill A. fumigatus spores in vivo. In addition, eosinopenia was correlated with higher mortality rates, decreased recruitment of inflammatory monocytes to the lungs, and decreased expansion of lung macrophages. As IL-17 signaling must occur on a local level to elicit its stereotypical response, such as the up-regulation of antimicrobial peptides and specific chemokines from stromal cells, eosinophils were discovered to be a significant source of pulmonary IL-17 as well as one of its upstream inducers, IL-23. In the context of asthma, this discovery opens a new paradigm whereby eosinophils might be driving asthma pathogenesis.

Introdução: Apesar dos recentes avanços no entendimento da fisiopatologia e no tratamento da espondilite anquilosante (EA), pouco se sabe acerca da influência das medicações anti-fator de necrose tumoral (anti-TNF) sobre as novas vias inflamatórias descritas na patogênese das espondiloartrites. Objetivo: Dessa forma, o objetivo desse estudo é investigar e descrever a influência a longo prazo das medicações anti-TNF sobre o eixo da IL-23/IL-17 em pacientes com EA e sua possível correlação com o tratamento, parâmetros clínicos, laboratoriais e radiológicos. Métodos: Oitenta e seis pacientes com EA sem exposição prévia a medicações anti-TNF foram recrutados. Desses, 47 possuíam Bath Ankylosing Spondylitis Disease Activity Index (BASDAI) >= 4 (grupo EA-ativo) e haviam sido encaminhados para iniciar tratamento anti-TNF e 39 possuíam BASDAI < 4 (grupo EA-controle) em uso de anti-inflamatório não hormonal (AINH) e/ou drogas antirreumáticas modificadoras do curso de doença tradicionais. O grupo EA-ativo foi avaliado clinicamente e laboratorialmente no tempo basal e após 12 e 24 meses de uso das medicações anti-TNF e foi comparado com o grupo EA-controle e com 47 controles saudáveis (CS) pareados por idade e sexo. O escore de uso de AINH foi calculado no tempo basal, 12 meses e 24 meses. Os níveis plasmáticos das interleucinas (IL) IL-17A, IL-22, IL-23 e PGE2 e a dosagem sérica da velocidade de hemossedimentação (VHS) e proteína C-reativa (PCR) foram realizados no grupo EA-ativo no tempo basal, 12 meses e 24 meses e somente no tempo basal nos grupos EA-controle e CS. No grupo EA-ativo, a progressão radiológica foi medida pelo modified Stoke Ankylosing Spondylitis Spine Score (mSASSS) no tempo basal e após 24 meses de tratamento. Resultados: No tempo basal, o grupo EA-ativo apresentou maiores níveis plasmáticos de IL-23 e PGE2 quando comparado ao grupo EA-controle (p < 0,001 e p=0,008) e ao grupo controle saudável (p < 0,001 e p=0,02). Após 24 meses de uso de anti-TNF, os níveis plasmáticos de IL-23 e PGE2 ainda se mantiveram elevados quando comparado ao grupo CS (p < 0,001 e p=0.03) apesar da melhora de todos os parâmetros clínicos e laboratoriais (VHS/PCR) (p < 0,001). A subanálise de 27 pacientes do grupo EA-ativo que obtiveram boa resposta ao uso de anti-TNF (atingiram ASDAS-PCR< 2,1 em 24 meses, com queda >= 1,1 em relação ao ASDAS-PCR basal) revelou que, ainda assim, os níveis plasmáticos de IL-23 eram superiores aos encontrados nos CS (p < 0,001) e superiores ao grupo EA-controle com atividade de doença similar (ASDAS-PCR < 2,1; p=0,01). No grupo EA-ativo foi encontrada uma correlação positiva entre os níveis plasmáticos de IL-23 e IL-17A no tempo basal, 12 meses e 24 meses do estudo (p <= 0,001). Conclusão: Os dados apresentados sugerem que o eixo da IL-23/IL-17 não é influenciado pelas medicações anti-TNF apesar da melhora dos parâmetros clínicos e marcadores de atividade inflamatória estudados / Background: Advances in pathophysiology and treatment of ankylosing spondylitis (AS) was recently demonstrated. However, the effect of anti-tumor necrosis fator (TNF) in the newly described inflammatory pathways involved in this disease remains to be determined. Objective: The aim of our study was, therefore, investigate long-term influence of anti-TNF drugs in IL-23/IL-17 axis of AS patients and their possible correlation with treatment, clinical, laboratory and radiographic parameters. Methods: Eighty six AS anti-TNF naïve patients, 47 referred for anti-TNF therapy (active-AS group; Bath Ankylosing Spondylitis Activity Index (BASDAI) >= 4) and 39 with BASDAI < 4 (control-AS group) were included. The active group was evaluated clinically and laboratorially at baseline, 12-months and 24-months after TNF blockade and compared at baseline to control-AS group and to 47 healthy age- and gender-matched controls. Plasma levels of interleukin (IL)17A, IL-22, IL-23 and PGE2 and serum levels of erythrocyte sedimentation rate (ESR) and c-reactive protein (CRP) were measured at three study times in active-AS and at baseline in control-AS and healthy-controls. Non-steroidal anti-inflammatory drugs (NSAIDs) intake were recorded at baseline, 12 months and 24 months. Radiographic severity and progression was assessed by modified Stoke Ankylosing Spondylitis Spine Score (mSASSS) at baseline and 24 months after therapy in active-AS patients. Results: At baseline, active-AS group presented higher IL-23 and PGE2 levels compared to control-AS group (p < 0.001 and p=0.008) and to healthy controls (p < 0.001 and p=0.02). After 24-months of TNF blockade, IL-23 and PGE2 remained elevated with higher levels compared with the healthy-control group (p < 0.001 and p=0.03) in spite of significant improvements in all clinical/inflammatory parameters (p < 0.001). Further analysis of 27 anti-TNF-treated patients who achieved a good response (ASDAS-CRP < 2.1, with a drop >= 1.1) at 24-months revealed that IL-23 plasma levels remained higher than healthy controls (p < 0.001) and higher than control-AS group with similar disease activity (ASDAS-CRP < 2.1, p=0.01). In active-AS group (n=47), there was a correlation between IL-23 and IL-17A at baseline, 12-months and 24-months after anti-TNF therapy (p <= 0.001). Conclusion: This study provides novel data demonstrating that the IL-23/IL-17 axis is not influenced by TNF blockade drugs in AS patients despite clinical and inflammation improvements and NSAID intake

Introdução: Apesar dos recentes avanços no entendimento da fisiopatologia e no tratamento da espondilite anquilosante (EA), pouco se sabe acerca da influência das medicações anti-fator de necrose tumoral (anti-TNF) sobre as novas vias inflamatórias descritas na patogênese das espondiloartrites. Objetivo: Dessa forma, o objetivo desse estudo é investigar e descrever a influência a longo prazo das medicações anti-TNF sobre o eixo da IL-23/IL-17 em pacientes com EA e sua possível correlação com o tratamento, parâmetros clínicos, laboratoriais e radiológicos. Métodos: Oitenta e seis pacientes com EA sem exposição prévia a medicações anti-TNF foram recrutados. Desses, 47 possuíam Bath Ankylosing Spondylitis Disease Activity Index (BASDAI) >= 4 (grupo EA-ativo) e haviam sido encaminhados para iniciar tratamento anti-TNF e 39 possuíam BASDAI < 4 (grupo EA-controle) em uso de anti-inflamatório não hormonal (AINH) e/ou drogas antirreumáticas modificadoras do curso de doença tradicionais. O grupo EA-ativo foi avaliado clinicamente e laboratorialmente no tempo basal e após 12 e 24 meses de uso das medicações anti-TNF e foi comparado com o grupo EA-controle e com 47 controles saudáveis (CS) pareados por idade e sexo. O escore de uso de AINH foi calculado no tempo basal, 12 meses e 24 meses. Os níveis plasmáticos das interleucinas (IL) IL-17A, IL-22, IL-23 e PGE2 e a dosagem sérica da velocidade de hemossedimentação (VHS) e proteína C-reativa (PCR) foram realizados no grupo EA-ativo no tempo basal, 12 meses e 24 meses e somente no tempo basal nos grupos EA-controle e CS. No grupo EA-ativo, a progressão radiológica foi medida pelo modified Stoke Ankylosing Spondylitis Spine Score (mSASSS) no tempo basal e após 24 meses de tratamento. Resultados: No tempo basal, o grupo EA-ativo apresentou maiores níveis plasmáticos de IL-23 e PGE2 quando comparado ao grupo EA-controle (p < 0,001 e p=0,008) e ao grupo controle saudável (p < 0,001 e p=0,02). Após 24 meses de uso de anti-TNF, os níveis plasmáticos de IL-23 e PGE2 ainda se mantiveram elevados quando comparado ao grupo CS (p < 0,001 e p=0.03) apesar da melhora de todos os parâmetros clínicos e laboratoriais (VHS/PCR) (p < 0,001). A subanálise de 27 pacientes do grupo EA-ativo que obtiveram boa resposta ao uso de anti-TNF (atingiram ASDAS-PCR< 2,1 em 24 meses, com queda >= 1,1 em relação ao ASDAS-PCR basal) revelou que, ainda assim, os níveis plasmáticos de IL-23 eram superiores aos encontrados nos CS (p < 0,001) e superiores ao grupo EA-controle com atividade de doença similar (ASDAS-PCR < 2,1; p=0,01). No grupo EA-ativo foi encontrada uma correlação positiva entre os níveis plasmáticos de IL-23 e IL-17A no tempo basal, 12 meses e 24 meses do estudo (p <= 0,001). Conclusão: Os dados apresentados sugerem que o eixo da IL-23/IL-17 não é influenciado pelas medicações anti-TNF apesar da melhora dos parâmetros clínicos e marcadores de atividade inflamatória estudados / Background: Advances in pathophysiology and treatment of ankylosing spondylitis (AS) was recently demonstrated. However, the effect of anti-tumor necrosis fator (TNF) in the newly described inflammatory pathways involved in this disease remains to be determined. Objective: The aim of our study was, therefore, investigate long-term influence of anti-TNF drugs in IL-23/IL-17 axis of AS patients and their possible correlation with treatment, clinical, laboratory and radiographic parameters. Methods: Eighty six AS anti-TNF naïve patients, 47 referred for anti-TNF therapy (active-AS group; Bath Ankylosing Spondylitis Activity Index (BASDAI) >= 4) and 39 with BASDAI < 4 (control-AS group) were included. The active group was evaluated clinically and laboratorially at baseline, 12-months and 24-months after TNF blockade and compared at baseline to control-AS group and to 47 healthy age- and gender-matched controls. Plasma levels of interleukin (IL)17A, IL-22, IL-23 and PGE2 and serum levels of erythrocyte sedimentation rate (ESR) and c-reactive protein (CRP) were measured at three study times in active-AS and at baseline in control-AS and healthy-controls. Non-steroidal anti-inflammatory drugs (NSAIDs) intake were recorded at baseline, 12 months and 24 months. Radiographic severity and progression was assessed by modified Stoke Ankylosing Spondylitis Spine Score (mSASSS) at baseline and 24 months after therapy in active-AS patients. Results: At baseline, active-AS group presented higher IL-23 and PGE2 levels compared to control-AS group (p < 0.001 and p=0.008) and to healthy controls (p < 0.001 and p=0.02). After 24-months of TNF blockade, IL-23 and PGE2 remained elevated with higher levels compared with the healthy-control group (p < 0.001 and p=0.03) in spite of significant improvements in all clinical/inflammatory parameters (p < 0.001). Further analysis of 27 anti-TNF-treated patients who achieved a good response (ASDAS-CRP < 2.1, with a drop >= 1.1) at 24-months revealed that IL-23 plasma levels remained higher than healthy controls (p < 0.001) and higher than control-AS group with similar disease activity (ASDAS-CRP < 2.1, p=0.01). In active-AS group (n=47), there was a correlation between IL-23 and IL-17A at baseline, 12-months and 24-months after anti-TNF therapy (p <= 0.001). Conclusion: This study provides novel data demonstrating that the IL-23/IL-17 axis is not influenced by TNF blockade drugs in AS patients despite clinical and inflammation improvements and NSAID intake

Ankylosing spondylitis (AS) is a common inflammatory arthritis of the spine and other affected joints, which is highly heritable, being strongly influenced by the HLA-B27 status, as well as hundreds of mostly unknown genetic variants of smaller effect. The aim of my research was to confirm some of the previously observed genetic associations and to identify new associations, many of which are in biological pathways relevant to AS pathogenesis, most notably the IL-23/T_H17 axis (IL23R) and antigen presentation (ERAP1 and ERAP2). Studies presented in this thesis include replication and refinement of several potential associations initially identified by earlier GWAS (WTCCC-TASC, 2007 and TASC, 2010). I conducted an extended study of IL23R association with AS and undertook a meta-analysis, confirming the association between AS and IL23R (non-synonymous SNP rs11209026, p=1.5 x 10-9, OR=0.61). An extensive re-sequencing and fine mapping project, including a meta-analysis, to replicate and refine the association of TNFRSF1A with AS was also undertaken; a novel variant in intron 6 was identified and a weak association with a low frequency variant, rs4149584 (p=0.01, OR=1.58), was detected. Somewhat stronger associations were seen with rs4149577 (p=0.002, OR=0.91) and rs4149578 (p=0.015, OR=1.14) in the meta-analysis. Associations at several additional loci had been identified by a more recent GWAS (WTCCC2-TASC, 2011). I used in silico techniques, including imputation using a denser panel of variants from the 1000 Genomes Project, conditional analysis and rare/low frequency variant analysis, to refine these associations. Imputation analysis (1782 cases/5167 controls) revealed novel associations with ERAP2 (rs4869313, p=7.3 x 10-8, OR=0.79) and several additional candidate loci including IL6R, UBE2L3 and 2p16.3. Ten SNPs were then directly typed in an independent sample (1804 cases/1848 controls) to replicate selected associations and to determine the imputation accuracy. I established that imputation using the 1000 Genomes Project pilot data was largely reliable, specifically for common variants (genotype concordence~97%). However, more accurate imputation of low frequency variants may require larger reference populations, like the most recent 1000 Genomes reference panels. The results of my research provide a better understanding of the complex genetics of AS, and help identify future targets for genetic and functional studies.