A neuroinflamação vem sendo estudada através da administração aguda, central ou periférica, de lipopolissacarídeos bacterianos (LPS) em roedores. A administração de LPS causa uma mudança na resposta imune do animal, aumentando os níveis de citocinas e outras quimiocinas inflamatórias. Essas alterações levam a um estado comportamental do tipo doente, que tende a ser extinto 24 h depois da administração de LPS, momento no qual se pode visualizar o desenvolvimento de um comportamento do tipo deprimido no animal. O hiperosídeo (HYP) é um flavonoide glicosilado, encontrado em diversas espécies vegetais e apresenta efeito do tipo antidepressivo, atividade citoprotetora, antioxidante e anti-inflamatória em roedores. O objetivo desse trabalho foi avaliar o efeito do flavonoide hiperosídeo (HYP) no modelo do comportamento de doente e do tipo deprimido induzidos pela administração de LPS em camundongos e sobre os níveis hipocampais de citocinas (IL-6, TNF, IFN-γ, IL-12 e IL-10) e quimiocina MCP-1. O modelo consistiu na exposição dos animais a uma sessão de 5 min de natação forçada (estímulo estressor) com posterior administração de LPS (600 μg/kg, i.p.) ou solução salina (10 mL/kg, i.p., SAL, animais controle do modelo). O comportamento de doente foi avaliado em campo aberto, através da medida do número de cruzamentos e score de sintomas, 06 e 24 h após a administração de LPS. O comportamento do tipo deprimido foi avaliado pela medida da imobilidade no teste de suspensão pela cauda (TSC) 24 h após a administração de LPS. Os tratamentos imipramina (IMI, 20 mg/kg), HYP (20 mg/kg) ou solução salina 10 mL/kg (grupo controle do tratamento) foram administrados (i.p.) 30 min antes (protocolo profilático) ou 5 h e 30 min depois (protocolo terapêutico) da administração de LPS ou SAL. Os níveis hipocampais de citocinas foram medidas por citometria de fluxo nos animais eutanasiados uma hora após o TSC. A administração profilática de HYP aumentou 12 os níveis de todas as citocinas tanto no grupo LPS quanto no grupo SAL. A análise da correlação citocinas pró-/anti-inflamatórias demonstrou que o tratamento profilático HYP resultou em uma correlação positiva IL-6/IL10 e TNF/IL-10 tanto nos animais SAL quanto LPS. A administração terapêutica de HYP aumentou somente os níveis de IL-10 e resultou em uma correlação IL-6/IL-10 positiva apenas no grupo SAL. Nos animais do grupo LPS, o tratamento terapêutico HYP aumentou os níveis de MCP-1 e resultou numa correlação positiva TNF/IL-10. O tratamento profilático HYP preveniu parcialmente o desenvolvimento do comportamento de doente, diminuindo o score de sintomas e o comportamento do tipo deprimido no grupo LPS. O tratamento terapêutico HYP preveniu o desenvolvimento do comportamento do tipo deprimido apenas no grupo SAL. A administração profilática de IMI nos animais SAL não alterou os níveis de IL-6, IL-10 e TNF, mas resultou em correlações IL-6/IL-10 e TNF/IL-10 positivas. Nos animais que receberam LPS, a administração profilática de IMI somente diminuiu os níveis de IL-6 e TNF, sem apresentar nenhuma correlação entre os níveis de citocinas. A administração terapêutica de IMI não alterou os níveis de nenhuma citocina, porém resultou em uma correlação TNF/IL-10 positiva. O tratamento profilático IMI preveniu tanto o desenvolvimento do comportamento de doente como do tipo deprimido. O tratamento terapêutico com IMI não reverteu os prejuízos induzidos pela administração de LPS, nem no comportamento de doente nem no do tipo deprimido. Em conclusão, os dados deste trabalho confirmam o efeito do tipo antidepressivo de hiperosídeo em camundongos e demonstram que o tratamento profilático com HYP tem efeito protetor parcial contra a manifestação dos sintomas de doente e comportamento do tipo deprimido induzidos pela natação forçada e administração de LPS. A ação do HYP pode estar relacionada ao aumento dos níveis hipocampais da citocina antiinflamatória IL-10. / The neuroinflammation has been studied through acute, central or peripheral administration of bacterial lipopolysaccharides (LPS) in rodents. LPS alters the animal immune response increasing the cytokines and others inflammatory chemokines levels. These alterations generate a group of symptoms and behavioral changes known as sickness behavior, which tends to be extinct 24 h after LPS administration when the animals develop a depressive-like behavior. Hyperoside (HYP) is a glycosylated flavonoid that present antidepressant, cytoprotective, antioxidant and anti-inflammatory properties in rodents. The aim of this study was to evaluate the HYP effects on sickness and depressive-like behaviors induced by LPS administration in mice as well as to investigate its effect on hippocampal cytokines (IL-6, TNF, IFN-γ, IL-12 e IL-10) and chemokine MCP-1 levels. The model consisted of a 5 min forced swimming session (stressor stimuli) with posterior LPS (600 μg/kg, i.p.) or saline (10 ml/kg, i.p., SAL) administration. The sickness behavior was assessed in the open field test by measuring the number of crossings and by scoring symptoms, 06 and 24 h after LPS administration. The depressive-like behavior was evaluated by measuring the immobility time in the tail suspension test (TST) 24 h after LPS administration. The treatments imipramine (IMI, 20 mg/kg), HYP (20 mg/kg) or saline 10 mL/kg (treatment control group) were administered (i.p.) 30 min before (prophylactic protocol) or 5 h and 30 min after (therapeutic protocol) LPS or saline (SAL - experiment control) administration. The cytokines levels were measured by flow cytometry in animals euthanized by decapitation 1 h after TST. The prophylactic administration of HYP increased all cytokines in both LPS and SAL groups, and resulted in an IL-6/IL-10 and a TNF/IL-10 positive correlation in both SAL and LPS animals. The therapeutic administration of HYP increased IL-10 levels, with an IL-6/IL10 positive correlation in SAL group only. In LPS animal group, the therapeutic 14 administration of HYP increased the MCP-1 levels and resulted in a TNF/IL-10 positive correlation. The prophylactic HYP did not prevent the reduction on spontaneous locomotion induced by LPS but decreased the symptoms score in LPS group and the depressive-like behavior in both LPS and SAL groups. The therapeutic HYP did not revert the sickness behavior and prevented the depressive-like behavior only in SAL group. The prophylactic IMI decreased IL-6 and TNF without any correlation between cytokines levels in animals that received LPS. In SAL animals, the prophylactic administration of IMI did not alter the cytokines levels but resulted in an IL-6/IL-10 and TNF/IL-10 positive correlations. The therapeutic IMI did not alter any cytokine levels, but resulted in a positive TNF/IL-10 correlation. The prophylactic IMI prevented both sickness and depressive-like behavior in LPS and SAL groups. The therapeutic IMI did not revert any LPS impairment. In conclusion, this study confirmed the antidepressant–like effect of hyperoside in mice and demonstrated that prophylactic treatment with this flavonoid partially protect the animals against sickness and depressive-like behaviors induced by LPS combined with forced swimming stress. Ability of HYP to increase the anti-inflammatory cytokine IL-10 hippocampal levels may underlies its effects on this model.

A morfina é um analgésico eficaz e muitas vezes opioide usado para aliviar a dor moderada a grave durante o período neonatal precoce. A exposição repetida de morfina no início da vida tem implicações duradouras para o desenvolvimento do sistema nervoso, tais como alterações neuroquímicas e comportamentais a longo prazo em ratos. O exercício físico vem sendo utilizado como uma alternativa não farmacológica para tratamento de quadros dolorosos. Deste modo nosso objetivo foi avaliar o efeito da exposição a morfina no período neonatal nas respostas nociceptiva (térmica e mecânica) e bioquímicas (citocinas e neurotrofinas) em ratos de P30 e P60 antes e após a exposição ao exercício físico. Ratos Wistar com 7 dias foram divididos em dois grupos: salina (SA) e morfina (MO) e submetidos a 5 mg / dia / 7 dias P8 para P14 a soro fisiológico ou MO respectivamente. Nas idades de P16, P30 e P60 a resposta nociceptiva térmica foi avaliada através do teste da placa quente (PQ), a resposta mecânica por Von Frey (VF) e Randal e Selitto (RS). Ainda foram medidos os níveis basais de BDNF, NGF, IL-6 e IL-10 em córtex cerebral e tronco encefálico. Após a sessão de exercício foram realizados em P30 e P60 o teste de PQ, 1h e 24h após o exercício, o teste de VF foi realizado 24h após e os níveis de BDNF, NGF, IL-6 e IL-10 também foram medidos em córtex cerebral e tronco encefálico após a exposição ao exercício. Nossos resultados demonstram que os animais que receberam morfina no período neonatal apresentam diminuição do limiar nociceptivo térmico e mecânico em P30 e P60. Os níveis de BDNF, NGF, IL-6 e IL-10 apresentaram relação direta com a idade em tronco encefálico, aumento ao longo do tempo. Em córtex cerebral os níveis de BDNF e NGF demonstraram uma interação entre os fatores grupo e idade, onde os animais do grupo MO têm diminuição desses com a idade. A IL-10 teve efeito somente da idade, enquando a IL-6 não se mostrou alterada por nenhum fator. Após a exposição ao exercício no teste da PQ no P30 e P60 os animais SAE tiveram uma diminuição do limiar nociceptivo se igualando aos grupos que receberam morfina. No teste de VF em P30 os grupos que receberam morfina são diferentes do grupos salina. Em P60 o grupo SAE mostra mais uma vez diminuiçao do limiar nociceptivo se igualando as grupos morfina. Nos níveis de BDNF e NGF em tronco encefálico ocorreu interação entre idade e grupo, onde o grupo MOE demonstra diminuição. Em tronco encefalico a IL-6 e Il-10 só tiveram efeito da idade. Em córtex cerebral os níveis de BDNF tiveram interação entre idade e grupo, o grupo MOE teve diminuição destes níveis em comparação aos demais grupos. Nos níveis de NGF se observou efeito do tempo e do grupo onde os grupos que recebem morfina têm níveis menores do que os que recebem salina em P60. O grupo MOS teve níveis menores de IL-6 em cortex cerebral do que os demais grupos, enquanto que os níveis de IL-10 só tiveram efeito da idade. Portanto a morfina no período neonatal leva a diminuição no limiar nociceptivo térmico e mecânico em ratos e que o exercício físico melhora os níveis BDNF, NGF e IL-6 em animais expostos a morfina no período neonatal. Porém o exercício físico não foi capaz de reverter a hiperalgesia e alodínia induzida pela morfina nos animais de P30 e P60. Sendo assim nossos dados mostram a necessidade de mais estudos sobre a dor em recém-nascidos e o sobre o uso de opioides neste período. Também se mostra necessário mais estudos sobre tratamentos não farmacológicos como o exercício físico. / Morphine is an effective analgesic often used to relieve moderate to severe pain during the early neonatal period. In our previous study, repeated morphine exposure in early life triggered persistent implications for the development of the nervous system, such as neurochemical and behavioral alterations in rats at long-term. The exercise has been used as a non-pharmacological alternative for treating painful conditions. Thus our aim was to evaluate the effect of repeated morphine exposure during the neonatal period upon nociceptive responses (thermal and mechanical) and biochemical markers (cytokines and neurotrophins) before and after unique physical exercise session in rats at P30 and P60. Seven-day-old male Wistar rats were divided into two groups: saline and morphine and subjected to saline and morphine (5 μg/day/7 days) from P8 to P14, respectively. At P16, P30 and P60, the thermal nociceptive response was assessed using the hot plate test (HP), while the mechanical response by Von Frey (VF) and Randal and Selitto (RS) tests. The basal levels of BDNF, NGF, IL-6 and IL-10 were measured in brainstem and cerebral cortex. One hour and 24h after exercise, the HP was conducted in P30 and P60, the VF test was only performed 24 ho after exercise, as well as the levels of BDNF, NGF, IL-6 and IL-10 were also measured in cerebral cortex and brainstem. Our results show that rats that received morphine in the neonatal period presented decreased thermal and mechanical nociceptive threshold in P30 and P60. And, BDNF, NGF, IL-6 and IL-10 levels presented a direct relationship with age in brainstem, increase their levels when the age increased. In cerebral cortex, BDNF and NGF levels showed an interaction between age and treatment group, where the morphine group showed decreased levels when the age increased. There was age effect upon IL-10 levels and no effects upon IL-6 levels in cerebral cortex. After 24h of exercise, saline group subjected to exercise presented decreased nociceptive threshold in using HP at P30 and P60, with similar threshold presented by morphine group. In VF test, both morphine groups presented decreased threshold in relation to both saline groups at P30. However, at P60, saline group subjected to exercise presented decreased nociceptive threshold, matching the morphine groups. In brainstem, we found interaction between age and group in BDNF and NGF levels, where morphine-exercise group showed decreased levels; and we observed only age effect upon IL-6 and IL-10 levels. In cerebral cortex, we observed interaction between age and group upon BDNF levels, where morphine-exercise group showed decreased levels compared to other groups. In relation to NGF levels, we observed effect of age and group, where morphine groups presented lower levels than saline groups in P60. The morphine-sedentary group presented lower IL-6 levels in the cerebral cortex than the other groups, while only age effect was observed on IL-10 levels. Our data lead us to conclude that morphine exposure in the neonatal period triggers a decrease in thermal nociceptive and mechanical thresholds in rats. And, the physical exercise improves BDNF, NGF and IL-6 levels in rats exposed to morphine in the neonatal period. However, on session of exercise was not able to revert the hyperalgesia and allodynia induced by morphine in rats at P30 and P60. Therefore, our data highlight the need of more studies about pain in newborns and neonates and the effect of the opioid use in this period. And, it is necessary more studies about non-pharmacological treatments, for example exercise.

Submitted by PPG Pediatria e Sa?de da Crian?a (pediatria-pg@pucrs.br) on 2018-08-30T19:28:55Z No. of bitstreams: 1 Disserta??o Final Gabriela Fabiano de Souza.pdf: 1295606 bytes, checksum: 81a673082dab9c871a133af1f8517135 (MD5) / Approved for entry into archive by Sheila Dias (sheila.dias@pucrs.br) on 2018-09-03T12:11:04Z (GMT) No. of bitstreams: 1 Disserta??o Final Gabriela Fabiano de Souza.pdf: 1295606 bytes, checksum: 81a673082dab9c871a133af1f8517135 (MD5) / Made available in DSpace on 2018-09-03T12:19:46Z (GMT). No. of bitstreams: 1 Disserta??o Final Gabriela Fabiano de Souza.pdf: 1295606 bytes, checksum: 81a673082dab9c871a133af1f8517135 (MD5) Previous issue date: 2018-03-19 / Conselho Nacional de Pesquisa e Desenvolvimento Cient?fico e Tecnol?gico - CNPq / Respiratory syncytial virus (RSV) is the major cause of infection in children up to five years of age. Reinfection is very common among patients, causing symptoms such as cold or allergy. However, in children, immunosuppressed patients and elderly infection is exacerbated leading to hospitalization and in some case, even death. The number of hospitalizations each year is alarming, even more so because up to now there is still no vaccine for RSV. Tissue damage in the lung caused by RSV leads to an immune response, where infected cells signal activation of signaling pathways, production of reactive oxygen species (ROS), and massive production of inflammatory mediators. Among this production is the macrophage migration inhibitory factor (MIF), which is a pro-inflammatory cytokine, which has been shown to play an important role in the immune response. Knowing this importance, we evaluated MIF expression macrophages from BALB/c mice. The cells were infected with different concentrations of RSV and analyzed by western blot, real-time PCR and Cytometric Bead Array (CBA). After confirmation of MIF expression by the infection, different inhibitors of signaling pathways and ROS were used to evaluate its importance for the expression of MIF. From the results obtained, we showed the dependence of ROS, 5-lipoxygenase (5-LOX), COX, PI3K and partially of P38 MAPK, for MIF expression, besides the need for viral activity. MIF was shown to be important for the release of cytokines such as TNF?, MCP-1 and IL-10. Based on this information MIF may play an important role in the exacerbation of infection, so it was extremely important to explore mechanisms involved in the expression of MIF in relation to RSV. / O V?rus sincicial respirat?rio (VSR) ? a maior causa de infec??o em crian?as at? os cinco anos de idade. A reinfec??o ? muito comum entre os pacientes, causando sintomas como de uma gripe ou alergia, no entanto, em crian?as, pacientes imunossuprimidos e idosos a infec??o ? muito mais exacerbada, o que acaba levando a necessidade de interna??o, podendo levar o paciente a ?bito. O n?mero de interna??es a cada ano ? alarmante, ainda mais que at? os dias atuais ainda n?o se tem uma vacina para o VSR. O dano tecidual no pulm?o, causado por VSR leva a uma resposta imune, onde c?lulas infectadas sinalizam para que ocorra a ativa??o de vias de sinaliza??o, produ??o de esp?cies reativas de oxig?nio (EROs) e tamb?m uma produ??o massiva de mediadores inflamat?rios. Dentre essa produ??o, est? o fator inibidor de migra??o de macr?fagos (MIF), que ? uma citocina pr?-inflamat?ria, que tem demonstrado um importante papel na resposta imune. Sabendo dessa import?ncia, avaliamos a express?o de MIF em macr?fagos de camundongos BALB/c. As c?lulas foram infectadas com diferentes concentra??es de VSR e analisadas por western blot, PCR em tempo real e Cytometric Bead Array (CBA). Ap?s a confirma??o da express?o de MIF pela infec??o, foram utilizados diferentes inibidores de vias de sinaliza??o e de EROs, para que fosse poss?vel avaliar sua import?ncia para a express?o de MIF. A partir dos resultados obtidos mostramos a depend?ncia de EROS, 5-lipoxigenase (5-LOX), COX, PI3K e parcialmente de P38 MAPK, para a express?o de MIF, al?m da necessidade de atividade viral. MIF se mostrou importante para a libera??o de citocinas como TNF ?, MCP-1 e IL-10. Baseado nessas informa??es MIF pode desempenhar um papel importante na exacerba??o da infec??o, sendo assim, foi de extrema import?ncia explorar mecanismos envolvidos na express?o de MIF em rela??o ao VSR.

<p>This study investigated the associations between IL-4 -590 C/T and IL-10 -1087 A/G polymorphisms and malariometric indexes in the Fulani and the Dogon ethnic groups living in sympatry in Mali and differing in susceptibility to malaria. The correlations between antibodies level and parasitological data as well as splenomegaly were assessed. The impact of IL-4 -590 variants on the levels of the studied antibodies was also studied. </p><p>The allele and genotype frequencies of both studied SNPs differed significantly between the two groups. The Fulani IL-4 T allele carriers had a significantly higher infection prevalence compared with those carrying the CC genotype. No correlation between anti-malarial antibody levels and parasite prevalence was seen in any of the communities. In the Fulani, the increase in total IgE levels was related to the presence of infection. Malaria-specific IgG4 levels were negatively correlated to the number of clones within the Fulani. The Fulani IL-4 T allele carriers had higher total and malaria-specific IgE levels, compared to the CC genotype carriers. These results suggest that the amount of antibodies may not be the key element in the protection against malaria. IgG4 might be involved in protection against malaria. The impact of IL-4 -590 variants on the antibody levels may be affected by other genetic/epigenetic/epistatic or environmental factors. </p><p>In the study in Senegal, multiplicity of infection (MOI) increased after the transmission season in all subjects, except in α-thalassaemic and in G6PD-mutated children, suggesting that α-thalassaemia may protect against infection by certain parasite strains. G6PD-mutated individuals may resist against increase in MOI after the transmission season due to rapid clearance of infection at an early stage. HbAs and the ABO system do not affect MOI in asymptomatic individuals. MOI was positively correlated to parasitemia, and did not vary over age (in the range of 2 to 10 years). No relation between MOI and clinical attack was noted. </p>

This study investigated the associations between IL-4 -590 C/T and IL-10 -1087 A/G polymorphisms and malariometric indexes in the Fulani and the Dogon ethnic groups living in sympatry in Mali and differing in susceptibility to malaria. The correlations between antibodies level and parasitological data as well as splenomegaly were assessed. The impact of IL-4 -590 variants on the levels of the studied antibodies was also studied. The allele and genotype frequencies of both studied SNPs differed significantly between the two groups. The Fulani IL-4 T allele carriers had a significantly higher infection prevalence compared with those carrying the CC genotype. No correlation between anti-malarial antibody levels and parasite prevalence was seen in any of the communities. In the Fulani, the increase in total IgE levels was related to the presence of infection. Malaria-specific IgG4 levels were negatively correlated to the number of clones within the Fulani. The Fulani IL-4 T allele carriers had higher total and malaria-specific IgE levels, compared to the CC genotype carriers. These results suggest that the amount of antibodies may not be the key element in the protection against malaria. IgG4 might be involved in protection against malaria. The impact of IL-4 -590 variants on the antibody levels may be affected by other genetic/epigenetic/epistatic or environmental factors. In the study in Senegal, multiplicity of infection (MOI) increased after the transmission season in all subjects, except in α-thalassaemic and in G6PD-mutated children, suggesting that α-thalassaemia may protect against infection by certain parasite strains. G6PD-mutated individuals may resist against increase in MOI after the transmission season due to rapid clearance of infection at an early stage. HbAs and the ABO system do not affect MOI in asymptomatic individuals. MOI was positively correlated to parasitemia, and did not vary over age (in the range of 2 to 10 years). No relation between MOI and clinical attack was noted.

African monkey species are natural hosts of simian hemorrhagic fever virus (SHFV) and develop persistent, asymptomatic infections. SHFV was previously shown to also cause a rapid onset fatal hemorrhagic fever disease in macaques. Infection of macaques with a new isolate of SHFV from persistently infected baboon sera, that showed high nucleotide identity with the lab strain LVR, resulted in viremia, pro-inflammatory cytokine and tissue factor production, and symptoms of coagulation defects. Primary macrophages and myeloid dendritic cell cultures from disease-susceptible macaques efficiently replicated SHFV and produced pro-inflammatory cytokines, including IL-6 and TNF-α, as well as tissue factor. Cells from disease resistant baboons produced low virus yields and the immunomodulatory cytokine IL-10. IL-10 treatment of macaque cells decreased IL-6 levels but had no effect on TNF-α levels, tissue factor or virus production suggesting that IL-10 plays a role in modulating immunopathology in disease-resistant baboons but not in regulating the efficiency of virus replication. SHFV is a member of the family Arteriviridae. The SHFV genome encodes 8 minor structural proteins. Other arteriviruses encode 4 minor structural proteins. Amino acid sequence comparisons suggest that the four additional SHFV minor structural proteins resulted from gene duplication. A full-length infectious clone of SHFV was constructed and produced virus with replication kinetics comparable to the parental virus. Mutant infectious clones, each with the start codon of one of the minor structural proteins substituted, were analyzed. All eight SHFV proteins were required for infectious virus production. The SHFV nonstructural polyprotein is processed into the mature replicase proteins by several viral proteases including papain-like cysteine proteases (PLPs). Only one or two PLP domains are present in other arteriviruses but SHFV has three PLP domains. Analysis of in vitro proteolytic processing of C- and N-terminally tagged polyproteins indicated that the PLP in each of the three SHFV nsp1 proteins is active. However, the nsp1α protease is more similar to a cysteine protease than a PLP. Analysis of the subcellular localization of the three SHFV nsp1 proteins indicated they have divergent functions.

Vitamin D is important for bone development with immunomodulatory effects. This study investigated whether feeding CD-1 and interleukin 10 (IL-10) knockout (KO) dams low (25 IU/kg diet) or high (5,000 IU/kg diet) vitamin D affected bone health of dams as well as their offspring. Offspring were weaned to 1 of the 2 diets and followed to young adulthood. Unlike CD-1 dams, IL-10 KO dams experienced greater femur strength with high vitamin D. CD-1 male offspring had reduced femur neck strength and female offspring had smaller, weaker femurs, and weaker lumbar vertebra 2 (LV2) with high maternal vitamin D. IL-10 KO male offspring had larger femurs and female offspring had stronger femurs when weaned to high vitamin D. Low vitamin D did not adversely impact bone health but the optimal level of dietary vitamin D seems to differ between healthy and inflammatory states.

Vitamin D is important for bone development with immunomodulatory effects. This study investigated whether feeding CD-1 and interleukin 10 (IL-10) knockout (KO) dams low (25 IU/kg diet) or high (5,000 IU/kg diet) vitamin D affected bone health of dams as well as their offspring. Offspring were weaned to 1 of the 2 diets and followed to young adulthood. Unlike CD-1 dams, IL-10 KO dams experienced greater femur strength with high vitamin D. CD-1 male offspring had reduced femur neck strength and female offspring had smaller, weaker femurs, and weaker lumbar vertebra 2 (LV2) with high maternal vitamin D. IL-10 KO male offspring had larger femurs and female offspring had stronger femurs when weaned to high vitamin D. Low vitamin D did not adversely impact bone health but the optimal level of dietary vitamin D seems to differ between healthy and inflammatory states.

La apoptosis es un proceso de muerte celular programada que está involucrada en la selección del repertorio de linfocitos T y en el mantenimiento de la tolerancia inmunológica, ya que es el mecanismo por el que se eliminan las células que podrían dar lugar a respuestas autoinmunes. Existen evidencias de que la alteración en los mecanismos apoptóticos están implicados en la patogenia y la actividad de las enfermedades autoinmunes sistémicas, de las cuales, el lupus eritematoso sistémico (LES) es la más representativa. Se han identificado en el ser humano algunos genes que codifican oncoproteínas y citocinas cuya transcripción parece ser crucial en este proceso: unos pro-apoptóticos (fas, p53 y TNF-alfa) y otros anti-apoptóticos (bcl-2 y IL-10). Se sospecha que cambios en la expresión de los mismos podrían desempeñar algún papel en la patogenia del LES, al favorecer la proliferación de determinadas poblaciones celulares de efecto autorreactivo.Con los trabajos de la presente tesis doctoral nos propusimos: determinar la implicación de los oncogenes (bcl-2, fas y p53) y las citocinas (IL-10 y TNF-alfa) en la disregulación apoptótica que presentan los pacientes con LES; estudiar la interrelación existente entre ellos; y analizar la relación de las posibles disregulaciones de los elementos que participan en la apoptosis con la actividad de la enfermedad lúpica.Los resultados obtenidos muestran cómo los oncogenes fas, bcl-2 y p53, las citocinas IL-10 y TNF-alfa, y la fracción proteica soluble del Fas (sFas) tienen una notable importancia en la patogenia y la actividad de la enfermedad lúpìca. Las vías apoptóticas del Fas y p53 son independientes entre sí. Sin embargo, diversas citocinas (IL-10, TNF-alfa), oncoproteínas (Bcl-2) y fracciones proteicas solubles (sFas) pueden ser las encargadas de relacionarlas entre sí. La interferencia de estas vías apoptóticas produciría una eliminación deficiente de los linfocitos autorreactivos. Ello favorecería su supervivencia, lo que provocaría las alteraciones de disregulación inmunológica propias del LES.

Autoimmune diseases are a group of different diseases which are characterized by an immune disorder leading to decreased tolerance to components of the body itself. These diseases have many factors that trigger such as a decrease of the share or percentage of regulatory T cells (Tregs). The Transforming Growth Factor-beta 1 (TGF-&#946;1) is involved in the suppression of the inflammatory response during the pathogenesis of autoimmune diseases (juvenile idiopathic arthritis, multiple sclerosis, diabetes), through the activation of this cell type. This cytokine is also associated with modulation of an inflammatory response either by increasing Treg cells and by modulating proinflammatory cytokines such as tumor necrosis factor alpha (TNF-&#945;). The components found in both innate immune responses as adaptive must be considered potential targets for developing new drugs immune modulators. Thus, manipulation of Tregs is an attractive strategy for immunotherapy and hence, the use of mimetic peptide to TGF-&#946;1 can be adopted to reduce the effects of severe autologous response, then creating an additional therapy for autoimmunity as well as for the treatment of inflammatory diseases. Our results show that we can select TGF-&#946;1 mimetic peptides since we can prove by bioinformatics both bind to this receptor molecule. Thus, the peptides can be used as immunomodulators to combat inflammation and in the treatment of autoimmune diseases since they can modulate the production of TNF-&#945; and IL-10. / Doenças autoimunes são um grupo de doenças distintas que se caracterizam por uma desordem imunológica levando a diminuição da tolerância aos componentes do próprio organismo. Essas doenças possuem vários fatores que as desencadeiam como a diminuição da ação ou porcentagem de células T regulatórias (Tregs). O Fator Transformante de Crescimento-beta 1 (TGF-&#946;1) está envolvido na supressão da resposta inflamatória durante a patogênese de doenças autoimunes (artrite idiopática juvenil, esclerose múltipla, diabetes), por meio da ativação desse tipo celular. Esta citocina também está associada a modulação de uma resposta inflamatória, seja pelo aumento de células Tregs como pela modulação de citocinas pro-inflamatórias como o Fator de necrose tumoral alfa (TNF-&#945;). Os componentes encontrados em respostas tanto imune inatas quanto adaptativas devem ser considerados potenciais alvos para o desenvolvimento de novos fármacos imuno moduladores. Assim, a manipulação de Tregs é uma estratégia atraente para a imunoterapia e, desta forma, o uso de peptídeos miméticos ao TGF-&#946;1 poderá ser adotado para diminuir as consequências de uma resposta autóloga severa, criando, então, uma terapia complementar para a autoimunidade bem como para o tratamento de doenças inflamatórias. Nossos resultados mostram que conseguimos selecionar peptídeos miméticos a molécula do TGF- &#946;1, uma vez que conseguimos provar por bioinformática que ambos se ligam ao receptor desta molécula. Assim, os peptídeos podem ser utilizados como imunomoduladores para o combate de inflamação e no tratamento de doenças autoimune já que conseguem modular a produção de TNF- &#945; e IL-10. Experimentos in vivo realizados também demonstraram a sua capacidade de diminuir inflamação modulação a migração de neutrófilos e leucócitos. / Mestre em Genética e Bioquímica