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Articular cartilage functions to reduce the mechanical stresses associated with diarthrodial joint movement, protecting these joints over a lifetime of use. Tissue function is maintained through the balance between synthesis and resorption (i.e., metabolism) of extracellular matrix (ECM) by articular chondrocytes (ACs). Two important hormonal regulators of cartilage metabolism are interleukin-1 (IL-1) and insulin-like growth factor-I (IGF-I). These factors have antagonistic effects on chondrocyte activity, and during the progression of osteoarthritis, IL-1 is thought to promote chondrocyte hyporesponsiveness to IGF-I. To better understand how the anabolic (IGF-I) and catabolic (IL-1) stimuli are linked within articular cartilage, we examined the mechanisms by which IL-1 regulates the IGF-I signaling system of ACs. Equine chondrocytes from non-arthritic stifle joints were multiplied over serial passages, re-differentiated in alginate beads, and stimulated with recombinant equine IL-1β. Chondrocytes were assayed for type I IGF receptor (IGF-IR), IGF binding proteins (IGFBPs), and endogenously-secreted IGF-I. Our experimental findings solidify the significance of IL-1 as a key regulator of IGF-I signaling within articular cartilage, demonstrating that regulation of the IGF-I system occurs through both direct (transcription) and indirect (proteolysis) mechanisms. These results have implications for molecular therapies (e.g., gene transfer) directed at reversing osteoarthritic cartilage deterioration. The presented research concerns not only cartilage biology but also tissue engineering strategies for cartilage repair. Alginate hydrogel culture has been reported to re-establish chondrocytic phenotype following monolayer expansion, but studies have not addressed effects on the signaling systems responsible for chondrocyte metabolism. We investigated whether chondrocyte culture history influences the IGF-I system and its regulation by IL-1. ACs expanded by serial passaging were either encapsulated in alginate beads or maintained on tissue culture plastic (TCP). Bead and TCP cells were plated at high-density, stimulated with IL-1β, and assayed for expression of IGF-I signaling mediators. Intermediate alginate culture yielded disparate basal levels of IGF-IR and IGFBP-2, which were attributed to differential transcription. The distinct mediator profiles coincided with varied effects of exogenous IL-1β and IGF-I on collagen Ia1 expression and cell growth rate. This study demonstrates that culture strategy impacts the IGF-I system of ACs, likely impacting their capability to mediate cartilage repair. / Ph. D.

De par sa présence dans tous les vaisseaux sanguins, l'endothélium joue un rôle clef dans le processus d’hémostase, tant par sa libération de facteurs anticoagulants que par ses changements protéiques qui permettent à l’organisme de déclencher la réparation tissulaire. La fonction anticoagulante de l’endothélium peut être mise en défaut en cas d’atteinte de son intégrité, entrainant la formation de thrombus, le rejet précoce de greffes ou encore l’induction de l’athérosclérose. L’intégrité de l’endothélium est donc capitale pour la prévention de nombreuses maladies cardiovasculaires. Chez l’adulte, les cellules endothéliales (CE), normalement quiescentes, sont rapidement activées en cas d’hypoxie ou d’inflammation, leur permettant ainsi d’amorcer le processus angiogénique comme suit: Tout d’abord, l’induction de l’hyperperméabilité vasculaire permet l’extravasation des protéines plasmatiques. Ensuite, la dégradation de la lame basale par des métalloprotéases permet aux CE de se détacher, de proliférer, de migrer et de s’organiser pour former l’ébauche du futur vaisseau. La dernière étape consiste en la maturation du vaisseau, c’est-à-dire son recouvrement par des cellules murales, telles que les cellules musculaires lisses et les péricytes. Ces processus sont régulés par de nombreux facteurs angiogéniques tels que les membres de la famille Notch, du vascular endothelial growth factor (VEGF), du fibroblast growth factor (FGF), des angiopoïétines, et des matrix metalloproteases (MMP). L’angiogenèse pathologique, soit une insuffisance ou un excès de vascularisation, est impliquée dans les blessures chroniques, les accidents cardiovasculaires, les pathologies coronariennes artérielles, les pathologies tumorales, l’arthrite rhumatoïde, la rétinopathie diabétique, l’athérosclérose, le psoriasis et l’asthme. Ces pathologies sont souvent issues d’une dérégulation de l’activité endothéliale, fréquemment observée conjointement à l’expression continue de molécules d’adhésion leucocytaires, à l’augmentation de la perméabilité vasculaire, et aux anomalies de la vasoréactivité. L’activation non-contrôlée de l’endothélium entraîne ainsi une inflammation chronique et la formation de structures vasculaires anarchiques. Les premiers leucocytes à répondre à l’appel inflammatoire sont les neutrophiles. Equippées d’une panoplie de produits antibactériens puissants mais aussi nocifs pour les tissus qui les entourent, ces cellules polylobées participent à chaque étape du processus inflammatoire, depuis l’induction de l’hyperperméabilité vasculaire jusqu’à la résolution. En effet, grâce à leurs récepteurs, les neutrophiles détectent et interprètent les signaux biochimiques présents dans la circulation et à la surface de l’endothélium, et libèrent aussi leurs propres médiateurs tels le VEGF, les MMP, et l’interleukine-8 (IL-8), dont les effets sont à la fois paracrines et autocrines. Existent-ils d’autres modulateurs typiques de la fonction endothéliale capables d’influencer le comportement des neutrophiles? En effet, notre laboratoire a démontré que chez l’humain, une stimulation directe aux angiopoïétines incitait les neutrophiles à adhérer aux CE, à migrer, à synthétiser et à relâcher l’IL-8, voire même à vivre plus longtemps. La présence du récepteur des angiopoïétines, Tie2, à la surface des neutrophiles laisse présager que la famille possèderait d’autres fonctions leucocytaires encore non-identifiées. Par ailleurs, dans un modèle classique de l’angiogenèse in vivo (matrigel), nous avons observé que sous l’effet du FGF1 et 2, les ébauches des nouveaux vaisseaux étaient parfois accompagnées d’une infiltration de cellules granulocytaires. Ainsi, en partant de ces observations, l’objectif de nos études (présentées ci-après) était d’approfondir nos connaissances sur la relation entre neutrophiles et facteurs angiogéniques, notamment les FGF et les angiopoïétines. Par tests in vitro, nous avons confirmé que les neutrophiles humains exprimaient plusieurs récepteurs du FGF (FGFR1-4) d’une façon hétérogène, et qu’ils migraient vers un gradient des ligands FGF1 et 2. Par ailleurs, nous nous sommes intéressés aux voies de signalisation inflammatoires activées par les ligands FGF1, FGF2, Ang1 et Ang2. Grâce à une stratégie génique ciblant 84 gènes inflammatoires, nous avons identifié plusieurs cibles d’intérêt touchées par Ang1, dont certains membres de la famille de l’IL-1, alors qu’aucun des gènes testés n’avait changé de façon significative sous l’effet des FGF ou d’Ang2. Suite à des cinétiques approfondies, nous avons démontré qu’Ang1 stimulait la transcription de l’ARN messager de l’IL-1β, et augmentait simultanément la quantité de protéine immature (pro-IL-1β; inactive) et clivée (IL-1β « mature »; active). En parallèle, Ang1 augmentait la sécrétion de l’antagoniste naturel de l’IL-1β, l’IL-1RA, sans pour autant stimuler la relâche de l’IL-1β. A l’instar des endotoxines bactériennes dont les effets liés à l’IL-1 dépendaient de la kinase p38, ceux d’Ang1 découlaient presque entièrement des voies de signalisation du p42/44. / Endothelial cells (ECs) form a monolayer that lines the inside of all blood vessels; thus, as the first barrier that separates blood elements from all things that fall beyond the blood vessel, ECs are strategically placed to play a central role in many essential physiological processes. While it is found mostly in a quiescent state in adult organisms, the endothelium retains a high level of plasticity that allows it to react to stimulus and dynamically control the passage of blood components to and from the bloodstream. For instance, upon detecting an activating angiogenic signal, ECs forgo their quiescence and undergo biochemical and structural changes necessary for the initiation of angiogenesis. Thus, activated ECs down-regulate their own expression of junctional molecules and secrete proteins to digest the extracellular matrix (ECM), thereby giving them the space to proliferate and migrate. Relaxing endothelial junctions also increases permeability, opening up the doorway for leukocyte infiltration. These cells can then modulate angiogenesis via their own set of mediators. Though the instigating stimuli may differ, the biochemical sequence of events that initiates angiogenesis is also common to the inflammatory response. In the latter case, changes in EC biochemistry include the release of chemotactic agents and expression of surface adhesion molecules, increasing the efficiency of leukocyte infiltration, particularly those of the myeloid lineage. Evidently, because angiogenesis and inflammation can be initiated by the same sequence of events, they will inevitably share effector molecules. Of the recruited leukocytes, neutrophils are generally the first responders at the site of inflammation, contributing mediators that propagate and eventually resolve inflammation. We and other groups have shown that endothelial modulators such as angiogenic growth factors exert a direct action on neutrophil activity independently of the presence of the endothelium. In particular, our laboratory has shown that members of the angiopoietin family and their receptor Tie2 are expressed by neutrophils and are capable of activating neutrophil intracellular signalling pathways that impact their survival, adhesion, migration, and protein production. The ability of angiopoietins to directly engage neutrophils illustrates an intimate link between angiogenesis and inflammation, and provides an explanation for why vascular pathologies are often accompanied by an exacerbated inflammatory response. In the studies presented herein, we sought to expand our understanding of the relationship between angiogenic growth factors and neutrophil behavior. In a pilot experiment using in vivo subcutaneous matrigel plugs, short-term treatment with fibroblast growth factors (FGF) 1 and 2 resulted in significant neovascularization; interestingly, the tissues surrounding the matrigel plug showed an increase in polymorphonuclear cell infiltration. Encouraged by the paucity of information in the literature regarding FGF-neutrophil interaction, we looked at the expression of FGF receptors (FGFRs) on neutrophils from different human donors, as well as the ability of FGFs to induce neutrophil chemotaxis. We demonstrated that the expression of FGFR was strongly dependent on genetic background: Overall, FGFR2 showed the highest incidence as a neutrophil cell-surface receptor, but none of the receptors were universally or uniformly expressed. Despite the genetic factor, neutrophils migrated in response to both FGF1 and FGF2 in vitro, suggesting that other neutrophil adaptors may be engaging FGFs. Given the shared ability of FGFs and angiopoietins (Ang) to induce neutrophil migration, we performed a wide-scale RNA assay to determine which genes were being engaged by the main ligands of both families. While none of FGF1, FGF2 or Ang2 had a strong effect on the 84 inflammatory cytokine genes tested (FGFs - unpublished data, 2011), at least two target genes belonging to the interleukin-1 (IL-1) family were significantly upregulated following Ang1 treatment. Further analysis showed that Ang1 not only stimulates gene transcription, but also translation and processing of the precursor of IL-1β (pro-IL-1β), and both precursor and mature proteins accumulate in the cell simultaneously. Interestingly, although no IL-1β is secreted from neutrophils after Ang1 or endotoxin (LPS) treatment, substantial quantities of the naturally occurring IL-1β antagonist (IL-1RA) are released, thereby tipping the balance in favor of inhibiting IL-1β activity. Finally, the activities of Ang1 on IL-1β and IL-1RA production and/or release are largely mediated by p42/44 MAPK; in contrast, the effects of LPS are driven by recruitment of p38.

4 ABSTRACT Interleukin-1α (IL-1α) is a pleiotropic cytokine and a key mediator of host immune response. It is synthesised as a 31-kDa precursor, that is cleaved by the cysteine protease calpain into the 17-kDa mature IL-1α and the 16-kDa N- terminal peptide of IL-1α (IL-1αNTP). Although IL-1α can be secreted, act on target cells through the surface receptor IL-1RI and trigger the signal transduction pathway, increasing evidence points toward the involvement of IL-1α in certain nuclear processes. IL-1αNTP is highly conserved among higher eukaryotes and contains a nuclear localisation sequence; indeed, both the precursor and IL-1αNTP are found in the cell nucleus. Previously, a genetic interaction of IL-1α with nuclear histone acetyltransferase (HAT) complexes has been reported from mammalian cells and, interestingly, also from the heterologous yeast model. This thesis extends the research of the nuclear function of IL-1α and demonstrates that IL-1α physically associates with the HAT/Core module of yeast SAGA and ADA HAT complexes. Results of the HAT subunit gene knock-out experiments followed by a set of co-immunoprecipitations also suggest a novel model of the yeast SAGA complex assembly, in which ADA appears to represent only a partly functional HAT complex. In its natural milieu of mammalian cells, IL-1α...

O uso de álcool está aumentando em nossa sociedade e permanece associado a inúmeros problemas sociais, econômicos e de saúde. De fato, o álcool e os problemas de saúde associados a ele exercem um impacto importante na prática médica e representam um dos maiores desafios da saúde pública. O consumo de álcool na sociedade contemporânea é geralmente aceito positivamente, dificultando o reconhecimento de certos padrões de consumo como doença. O alcoolismo é um transtorno recidivante crônico caracterizado pela ingestão compulsiva de quantidades excessivas de etanol, perda de controle em sua ingestão, comportamento inadequado e a presença de um estado emocional negativo. O consumo de quantidades nocivas de álcool resulta em danos físicos e psicológicos e o vicio é um transtorno psiquiátrico que afeta as funções executivas, causando perda de interesse em outros alvos do prazer e comportamento compulsivo de busca por drogas. O álcool interage com vários sistemas neurológicos. O presente trabalho analisou a resposta imunoinflamatória no tecido cerebral de camundongos CRAMP knockout (KO) jovens e tipo selvagem (WT) submetidos ao modelo de intoxicação alcoólica, com o objetivo de investigar o impacto de CRAMP na dependência alcoólica no adolescente. O CRAMP é um peptídeo antimicrobiano com efeitos pleotrópicos e, até onde sabemos, seu papel nunca foi investigado nesse sentido. Também analisamos a secreção de vários neuropeptídeos, proteínas e citocinas. Nossos resultados mostraram uma diferença significativa na ingestão de etanol entre os animais comparados CRAMP KO e WT, o que foi relacionado a um aumento nos níveis cerebelares de IL-1beta. Concluimos que os pepitídeos antimicrobianos podem ter um papel importante na resposta imunoinflamatória que ocorre durante o etilismo agudo / The use of alcohol is increasing in our society and remains associated with countless social, economic and health problems. In fact, alcohol and the health issues associated to its abuse exert an important impact on medical practice and represent one of the biggest challenges of public health. The consumption of alcohol in contemporary society is generally accepted positively, making certain patterns of consumption very difficult to be recognized as a disease. Alcoholism is a chronic relapsing disorder characterized by compulsive ingestion of excessive amounts of ethanol, loss of control in its intake, inappropriated behavior and the presence of a negative emotional state. The consumption of harmful amounts of alcohol results in physical and or psychological damage and addiction is a psychiatric disorder that affects the executive functions, causing loss of interest in other aspects of life and a compulsive behavior. Alcohol interacts with several neurologic systems. The present work analyzed the immuno-inflammatory response in the brain tissue of young CRAMP knockout (KO) and wild-type (WT) mice submitted to a model of alcohol intoxication, in order to investigate the impact of CRAMP in teenager alcohol addiction. CRAMP is an antimicrobial peptide with pleotropic effects and, as far as we know, its role had never been investigation in this regard. We also analysed the secretion of several neuropeptides, proteins and cytokines. Our results showed a significant difference in ethanol intake when CRAMP KO and WT animals were compared, which was related to an increase in the cerebellar levels of IL-1beta. We conclude that antimicrobial peptides may play an important role in the immunoinflammatory response that occurs during acute alcoholism

A candidíase vulvovaginal corresponde a uma das mais frequentes infecções do trato reprodutivo. Estima-se que 75% das mulheres na idade reprodutiva experimentarão pelo menos um episódio de candidíase vulvovaginal durante suas vidas, a maioria evoluirá com episódios infrequentes, entretanto, 5% sofrerão recorrência, ou seja, quatro ou mais episódios de candidíase vulvovaginal comprovadas clínica e laboratorialmente no período de 1ano. Os mecanismos pelos quais as recorrências ocorrem ainda são pouco conhecidos, estando provavelmente relacionados à alterações na imunidade local. O presente estudo teve como objetivo avaliar as associações entre os polimorfismos nos genes que codificam a lecitina ligadora de manose (MBL) e do antagonista do receptor da interleucina 1 (IL1-Ra) com a candidíase vulvovaginal recorrente (CVVR) em mulheres brasileiras. Foram estudadas 100 mulheres portadoras de CVVR atendidas no Serviço de Imunologia Genética e Infecções do Trato Reprodutivo da Disciplina de Ginecologia da Faculdade de Medicina da Universidade de São Paulo. Para a análise dos polimorfismos nos genes que codificam para a MBL e o IL1-Ra realizou-se coleta de células bucais que foram enviadas para Division of Immunology and Infectious Diseases of Weill Medical College of Cornell University Resultados: Mulheres com candidíase vulvovaginal recorrente apresentaram maior frequência de polimorfismo no códon 54 do gene que codifica a MBL quando comparadas a mulheres saudáveis. Não foram observadas diferenças estatisticamente significativas na frequência do polimorfismo do gene que codifica o IL1-Ra entre os grupos estudados / Vulvovaginal candidiasis is the most common genital infection in women during their childbearing years. About 75% of women suffer at least one syntomatic episode during their lives. Most of them will have infrequent episodes, but 5% will suffer recurrent episode of vulvovaginal candidiasis. The mechanisms responsible for recurrent vulvovaginal candidiasis (RVCC) remain a matter of speculation, although an alteration in local immunity appears to be a major factor. The aim of this study was to assess the correlation between polymorphisms in the genes coding for mannose-binding lectin (MBL) and interleukin-1 receptor antagonist (IL1-Ra) and RVCC in women from São Paulo, Brazil. The study population consisted of 100 women with RVCC, who were seen at Serviço de Infecções do Trato Reprodutivo da Disciplina de Ginecologia da Faculdade de Medicina da Universidade de São Paulo. To analyse for the MBL códon 54 gene polymorphism and for IL1-Ra, buccal cells were obtained with a cotton swabs and shipped to New York at ambient temperature. The polymorphisms were identified in the Division of Immunology and Infectious Diseases of Weill Medical College of Cornell University. Results: Women with RVVC present a high frequency of polymorphisms at codon 54 in the gene coding for MBL; on the other hand there were no differences in polymorphism frequency in the gene coding for IL1-Ra when compared to control women

Embora já tenha sido comprovada a ação adjuvante da SBA-15, pouco se sabe sobre o seu mecanismo molecular que leva a modulação positiva da resposta imunológica. Foi avaliada a ativação do inflamassoma NLRP3, sobre estímulos de SBA-15, em macrófagos de camundongos C57BL/6. Como parâmetro dessa ativação, foi analisada a produção de IL-1β por ELISA. A SBA-15 foi capaz de induzir a produção de IL-1β a níveis semelhantes quando comparado com um agonista de NLRP3 (Nano-SiO2), sugerindo a ativação do inflamassoma. Para avaliar o envolvimento da caspase-1, nos resultados obtidos com a SBA-15, os macrófagos foram estimulados com sílica na presença do inibidor de caspase-1, e como esperado, a produção de IL-1β foi restaurada para o seu nível basal. A ativação do inflamassoma, por estímulos da SBA-15, parece ser parcialmente dependente da fagocitose e da produção das espécies reativas do oxigênio. Além disso, foi visto que a SBA-15 não induz a produção de IL-6, confirmando que essa sílica está envolvida na via do inflamassoma e não em outras vias, como por exemplo, NF-κB. / Although it has already been proven adjuvant action of SBA-15, little is known about its molecular mechanism leading to positive modulation of the immune response. NLRP3 inflammasome activation was evaluated on SBA-15 stimulation in C57BL/6 mice macrophages. As this parameter activation, it analyzed the production of IL-1β by ELISA. The SBA-15 was able to induce the production of IL-1β at levels similar when compared to an agonist of NLRP3 (Nano-SiO2), suggesting the activation of the inflammasome. To assess the involvement of caspase-1, the results obtained with SBA-15, the macrophages were stimulated with silica in the presence of caspase-1 inhibitor, and as expected, IL-1β production was restored to its baseline level. Activation of the inflammasome, by stimuli of SBA-15, appears to be partly dependent on phagocytosis and production of reactive oxygen species. In addition, it was seen that the SBA-15 does not induce IL-6 production, confirming that silica is involved inflammasome the path of and not in other ways, eg, NF-κB.

A periodontite agressiva (PAg) compreende um grupo de doenças periodontais caracterizadas por rápida destruição dos tecidos periodontais, em indivíduos jovens e que geralmente não apresentam doenças sistêmicas. Estudos em populações e em famílias indicaram que fatores genéticos possuem influência na susceptibilidade à PAg. Os polimorfismos genéticos da interleucina-1 (IL-1) e do fator de necrose tumoral-? (TNF-?) foram associados com o aumento da severidade da periodontite. O objetivo deste estudo foi avaliar a associação entre o polimorfismo dos genes IL-1A (-889), IL-1B (-511), IL-1B (+3954), IL-1RN (íntron 2-VNTR) e TNFA (-308) e a PAg. O DNA genômico foi extraído de 150 indivíduos não fumantes, sendo 50 portadores de PAg e 100 indivíduos com periodonto saudável. O polimorfismo dos genes IL-1A (-889), IL-1B (-511), IL-1B (+3954) e TNFA (-308) foi analisado utilizando-se a técnica da reação em cadeia da polimerase e análise do polimorfismo de comprimento dos fragmentos de restrição (PCR-RFLP). O polimorfismo de número variável de repetições em tandem (VNTR) no íntron 2 do gene IL-1RN foi detectado pela técnica da PCR e análise do tamanho dos fragmentos. A análise estatística revelou que não houve diferença significante na freqüência dos genótipos e alelos, entre o grupo com PAg e os indivíduos com periodonto saudável, para IL-1A (-889), IL-1B (-511), IL-1B (+3954) e TNFA (-308) (p-valor>0.05%). Entretanto, houve diferença estatisticamente significante, entre os dois grupos, na distribuição dos genótipos para IL-1RN (íntron 2-VNTR) (teste Exato de Fisher; p-valor=0,021) e alelos (teste Exato de Fisher; p-valor=0,04). Os achados sugerem que o polimorfismo do gene IL-1RN (íntron 2-VNTR) está associado com a PAg na população estudada. / Aggressive periodontitis (AgP) is a type of periodontal disease characterized by a rapid destruction of the periodontal supporting tissues in young adults who are usually systemically well. Population and family studies indicate that genetic factors seem to have a strong influence on susceptibility to AgP. Genetic polymorphism at the interleukin-1 (IL-1) and tumor necrosis factor alpha (TNFA) were associated with the increase on the severity of periodontitis. This study aimed at finding a possible association between IL-1A (-889), IL-1B (-511), IL-1B (+3954), IL-1RN (intron 2-VNTR) and TNFA (-308) polymorphism in patients with aggressive periodontitis. Genomic DNA was extracted from the saliva of 150 nonsmoking subjects, 50 patients with AgP and 100 periodontal healthy subjects. All individuals were systemically healthy. The polymorphisms of IL-1A (-889), IL-1B (-511), IL-1B (+3954) and TNFA (-308) were analyzed by means of polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). The polymorphism of a variable number tandem repeat (VNTR) in intron 2 of the IL-1RN gene was detected by PCR amplification and fragment size analysis. Statistical analysis revealed no significant difference in the IL-1A (-889), IL-1B (-511), IL-1B (+3954) and TNFA (-308) genotypes and allele frequencies between AgP patients and periodontal healthy subjects (p-value>0.05%). However, there were significant differences among the groups in the distribution of IL-1RN (íntron 2-VNTR) genotypes (Fisher\'s exact test; p-value=0.021) and allele frequencies (Fisher\'s exact test; p-value=0.04). These findings suggest that IL-1RN (intron 2-VNTR) polymorphism is associated with AgP in the population presented here.

A periodontite agressiva (PAg) compreende um grupo de doenças periodontais caracterizadas por rápida destruição dos tecidos periodontais, em indivíduos jovens e que geralmente não apresentam doenças sistêmicas. Estudos em populações e em famílias indicaram que fatores genéticos possuem influência na susceptibilidade à PAg. Os polimorfismos genéticos da interleucina-1 (IL-1) e do fator de necrose tumoral-? (TNF-?) foram associados com o aumento da severidade da periodontite. O objetivo deste estudo foi avaliar a associação entre o polimorfismo dos genes IL-1A (-889), IL-1B (-511), IL-1B (+3954), IL-1RN (íntron 2-VNTR) e TNFA (-308) e a PAg. O DNA genômico foi extraído de 150 indivíduos não fumantes, sendo 50 portadores de PAg e 100 indivíduos com periodonto saudável. O polimorfismo dos genes IL-1A (-889), IL-1B (-511), IL-1B (+3954) e TNFA (-308) foi analisado utilizando-se a técnica da reação em cadeia da polimerase e análise do polimorfismo de comprimento dos fragmentos de restrição (PCR-RFLP). O polimorfismo de número variável de repetições em tandem (VNTR) no íntron 2 do gene IL-1RN foi detectado pela técnica da PCR e análise do tamanho dos fragmentos. A análise estatística revelou que não houve diferença significante na freqüência dos genótipos e alelos, entre o grupo com PAg e os indivíduos com periodonto saudável, para IL-1A (-889), IL-1B (-511), IL-1B (+3954) e TNFA (-308) (p-valor>0.05%). Entretanto, houve diferença estatisticamente significante, entre os dois grupos, na distribuição dos genótipos para IL-1RN (íntron 2-VNTR) (teste Exato de Fisher; p-valor=0,021) e alelos (teste Exato de Fisher; p-valor=0,04). Os achados sugerem que o polimorfismo do gene IL-1RN (íntron 2-VNTR) está associado com a PAg na população estudada. / Aggressive periodontitis (AgP) is a type of periodontal disease characterized by a rapid destruction of the periodontal supporting tissues in young adults who are usually systemically well. Population and family studies indicate that genetic factors seem to have a strong influence on susceptibility to AgP. Genetic polymorphism at the interleukin-1 (IL-1) and tumor necrosis factor alpha (TNFA) were associated with the increase on the severity of periodontitis. This study aimed at finding a possible association between IL-1A (-889), IL-1B (-511), IL-1B (+3954), IL-1RN (intron 2-VNTR) and TNFA (-308) polymorphism in patients with aggressive periodontitis. Genomic DNA was extracted from the saliva of 150 nonsmoking subjects, 50 patients with AgP and 100 periodontal healthy subjects. All individuals were systemically healthy. The polymorphisms of IL-1A (-889), IL-1B (-511), IL-1B (+3954) and TNFA (-308) were analyzed by means of polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). The polymorphism of a variable number tandem repeat (VNTR) in intron 2 of the IL-1RN gene was detected by PCR amplification and fragment size analysis. Statistical analysis revealed no significant difference in the IL-1A (-889), IL-1B (-511), IL-1B (+3954) and TNFA (-308) genotypes and allele frequencies between AgP patients and periodontal healthy subjects (p-value>0.05%). However, there were significant differences among the groups in the distribution of IL-1RN (íntron 2-VNTR) genotypes (Fisher\'s exact test; p-value=0.021) and allele frequencies (Fisher\'s exact test; p-value=0.04). These findings suggest that IL-1RN (intron 2-VNTR) polymorphism is associated with AgP in the population presented here.

A candidíase vulvovaginal corresponde a uma das mais frequentes infecções do trato reprodutivo. Estima-se que 75% das mulheres na idade reprodutiva experimentarão pelo menos um episódio de candidíase vulvovaginal durante suas vidas, a maioria evoluirá com episódios infrequentes, entretanto, 5% sofrerão recorrência, ou seja, quatro ou mais episódios de candidíase vulvovaginal comprovadas clínica e laboratorialmente no período de 1ano. Os mecanismos pelos quais as recorrências ocorrem ainda são pouco conhecidos, estando provavelmente relacionados à alterações na imunidade local. O presente estudo teve como objetivo avaliar as associações entre os polimorfismos nos genes que codificam a lecitina ligadora de manose (MBL) e do antagonista do receptor da interleucina 1 (IL1-Ra) com a candidíase vulvovaginal recorrente (CVVR) em mulheres brasileiras. Foram estudadas 100 mulheres portadoras de CVVR atendidas no Serviço de Imunologia Genética e Infecções do Trato Reprodutivo da Disciplina de Ginecologia da Faculdade de Medicina da Universidade de São Paulo. Para a análise dos polimorfismos nos genes que codificam para a MBL e o IL1-Ra realizou-se coleta de células bucais que foram enviadas para Division of Immunology and Infectious Diseases of Weill Medical College of Cornell University Resultados: Mulheres com candidíase vulvovaginal recorrente apresentaram maior frequência de polimorfismo no códon 54 do gene que codifica a MBL quando comparadas a mulheres saudáveis. Não foram observadas diferenças estatisticamente significativas na frequência do polimorfismo do gene que codifica o IL1-Ra entre os grupos estudados / Vulvovaginal candidiasis is the most common genital infection in women during their childbearing years. About 75% of women suffer at least one syntomatic episode during their lives. Most of them will have infrequent episodes, but 5% will suffer recurrent episode of vulvovaginal candidiasis. The mechanisms responsible for recurrent vulvovaginal candidiasis (RVCC) remain a matter of speculation, although an alteration in local immunity appears to be a major factor. The aim of this study was to assess the correlation between polymorphisms in the genes coding for mannose-binding lectin (MBL) and interleukin-1 receptor antagonist (IL1-Ra) and RVCC in women from São Paulo, Brazil. The study population consisted of 100 women with RVCC, who were seen at Serviço de Infecções do Trato Reprodutivo da Disciplina de Ginecologia da Faculdade de Medicina da Universidade de São Paulo. To analyse for the MBL códon 54 gene polymorphism and for IL1-Ra, buccal cells were obtained with a cotton swabs and shipped to New York at ambient temperature. The polymorphisms were identified in the Division of Immunology and Infectious Diseases of Weill Medical College of Cornell University. Results: Women with RVVC present a high frequency of polymorphisms at codon 54 in the gene coding for MBL; on the other hand there were no differences in polymorphism frequency in the gene coding for IL1-Ra when compared to control women