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.

Submitted by Simone Souza (simonecgsouza@hotmail.com) on 2017-09-18T15:03:59Z No. of bitstreams: 1 DISS_2014_Caroline Marques Caloi.pdf: 1667299 bytes, checksum: 818ef12f2884a1595534e6fe5023eb67 (MD5) / Approved for entry into archive by Jordan (jordanbiblio@gmail.com) on 2017-09-22T11:54:23Z (GMT) No. of bitstreams: 1 DISS_2014_Caroline Marques Caloi.pdf: 1667299 bytes, checksum: 818ef12f2884a1595534e6fe5023eb67 (MD5) / Made available in DSpace on 2017-09-22T11:54:23Z (GMT). No. of bitstreams: 1 DISS_2014_Caroline Marques Caloi.pdf: 1667299 bytes, checksum: 818ef12f2884a1595534e6fe5023eb67 (MD5) Previous issue date: 2014-09-22 / A hanseníase é uma doença infecciosa crônica, causada pelo Mycobacterium leprae (M. leprae), parasita intracelular obrigatório que afeta, preferencialmente, macrófagos no tecido conectivo e células de Schwann nos nervos periféricos. O objetivo desse trabalho foi identificar as células mnonucleares presentes na pele de pacientes com hanseníase, com as formas clínicas tuberculóide-tuberculóide (TT), tuberculóide-borderline (TB), borderline-borderline (BB), virchowiano-borderline (BV) e virchowiano-virchowiano (VV); e mensurar a expressão dos mediadores anti-inflamatórios anexina-A1 (ANXA1) e interleucina-10 (IL-10), investigando seu possível papel na patogênese da doença. Após o diagnóstico clínico os pacientes foram submetidos à biópsia da lesão para análise histopatológica. Os cortes histopatológicos foram submetidos à técnica de imunofluorescência para detecção dos mediadores ANXA1 e IL-10. Os resultados demonstraram uma correlação na diferença dos tipos de leucócitos presentes nas diferentes formas clínicas dos pacientes com hanseníase com os níveis dos mediadores ANXA1 e IL-10 produção diferencial de mediadores reguladores do processo infeccioso. Os dados demonstraram que os histiócitos, células T CD4+ e T CD8+ dos pacientes com as formas clínicas BV e VV apresentavam uma expressão aumentada da proteína ANXA1, assim como nos níveis da citocina IL-10. Além disso, pode-se verificar uma correlação positiva na expressão da ANXA1 e nos níveis de IL-10 nos pacientes BB, BV e VV, indicando possivelmente que essa proteína possa contribuir com a regulação da produção dessa citocina durante o processo infeccioso induzido por M. leprae. Em conclusão, a proteína ANXA1 foi caracterizada como um possível regulador do processo infeccioso induzido por M. leprae, indicando que, a presença dessa proteína poderia estar reduzindo o potencial pró-inflamatório nos pacientes BB, BV e VV, levando a disseminação do M. leprae. Do contrário, em pacientes TT e BT, a proteína ANXA1 apresenta-se reduzida, o que poderia contribuir com a ativação dos leucócitos e a eliminação das bactérias. / Leprosy is a chronic infectious disease caused by Mycobacterium leprae (M. leprae), a intracellular obligate parasite that affects mainly macrophages and Schwann cells. This study’s goal was to identify monocytic cells present in the skin of leprosy patients with tuberculoid- tuberculoid clinical forms (TT), tuberculoid-borderline (TB), borderline-borderline (BB), borderline-lepromatous (BV) and lepromatous (LL); and evaluating the expression of annexin-A1 (ANXA1), and anti-inflammatory mediators interleukin-10 (IL-10), evaluating its potential role in pathogenesis disease. After the clinical diagnosis, the patients underwent biopsy for histopathological analysis. Histopathological cuts were submitted to immunofluorescence staining for detection of mediators ANXA1 and IL-10. The results showed a difference in the of leukocytes’s profile in patients with different clinical forms of leprosy, that is essential to allow the differential production of regulatory mediators of the infectious process. The data showed that the histiocytes, CD4 + and CD8 + T cells of patients with clinical forms BV and VV had a higher expression of ANXA1 protein as well as high levels of IL-10. Furthermore, it can be seen a positive correlation in expression of ANXA1 and IL-10 in BB, BV and VV patients, possibly indicating that this protein may contribute to the regulation of this cytokine production during the infectious process induced by M. leprae. In conclusion, ANXA1 protein was characterized as a possible regulator of the infectious process induced by M. leprae, indicating that the presence of this protein could be reducing the pro-inflammatory potential in BB, BV and VV patients taking the spread of the bacterium M. leprae. Otherwise, patients in TT and BT, ANXA1 protein comes in lower levels, which could contribute to more effective activation of pro-inflammatory action of the immune system, favoring the elimination of bacterias.

Analisamos a expressão de IL-10, IFN-<font face=\"symbol\">g e, seus receptores pelas células trofoblásticas de camundongos, citocinas anti e pró-inflamatórias. Cones ectoplacentários aos 7,5 dias de gestação foram cultivados por 48 h e em seguida tratados com 100 U/mL IFN-<font face=\"symbol\">g ou 10 <font face=\"symbol\">hg/mL IL-10. Após 6 h e 14 h, as amostras foram processadas para análise da expressão gênica por RT-PCR e protéica por imunohistoquímica, respectivamente. Grupos controle não receberam tratamento. IFN-<font face=\"symbol\">g aumentou a expressão de IL-10R1 mas não a de IL-10 nas células trofoblásticas. IL-10, ao contrário, aumentou a expressão de IFN-<font face=\"symbol\">g, mas diminuiu IFN-<font face=\"symbol\">gR<font face=\"symbol\">a e não alterou IFN-<font face=\"symbol\">gR<font face=\"symbol\">b. Reações imunohistoquímicas confirmaram os resultados de expressão gênica. Isto sugere que o trofoblasto pode participar da imunidade da interface materno-placentária aumentando a expressão de IFN-<font face=\"symbol\">g em situações em que no meio há aumento de citocinas anti-inflamatórias, o que deve ser o reflexo da necessidade e importância desta citocina para o sucesso da gestação. / Key cytokines such as IL-10 and IFN-<font face=\"symbol\">g, essential for immune response regulation, have also been found locally at maternal-placental interface during mouse pregnancy. Particularly, levels of IL-10 characterize an anti-inflammatory environment associated to the inhibition of T helper-1 lymphocytes (Th1) development and the proliferative stimulation of the B lymphocytes (humoral response). On the other hand, increases in IFN-<font face=\"symbol\">g profile prevent T helper-2 lymphocytes (Th2) activation leading to an inflammatory response that favors a Th1 response. The local production of these cytokines by NK uterine cells and T gd lymphocytes are relevant, but not exclusive. Thus, this study analyzed the potential contribution of the trophoblast in the maintenance of Th1/Th2 balance in the maternal-placental interface, represented, respectively by the expression of IL-10 and IFN-<font face=\"symbol\">g cytokines. The expression of the anti-inflammatory cytokine IL-10 and its receptor was evaluated in the presence of an inflammatory environment mimetized by IFN-<font face=\"symbol\">g addition to the culture medium. On contrary, the expression of the IFN-<font face=\"symbol\">g (and its receptor) was evaluated in the presence of IL-10 characterizing an anti-inflammatory condition. Mouse trophoblast cells were isolated from implantation sites at gestational day 7.5 and cultured in standard conditions. Gene and protein expression were determined by immunohistochemistry and RT-PCR. IL-10 and IFN-<font face=\"symbol\">g and their receptors were expressed in cultured trophoblast cells in the absence or presence of IFN-<font face=\"symbol\">g and IL-10, respectively. IFN-<font face=\"symbol\">g treatment increased IL-10R1 expression but do not alter IL-10 expression. On contrary, in the presence of IL-10 the IFN-<font face=\"symbol\">g expression increased significantly while the expression of its receptor decreased. These results suggest that a proinflammatory environment increases trophoblast responsiveness to IL-10 whereas an anti-inflammatory condition seems to reinforce the importance of IFN-<font face=\"symbol\">g expression at the maternal-placental interface, on the initial periods of gestation.

Submitted by Renata Lopes (renatasil82@gmail.com) on 2016-10-05T11:27:00Z No. of bitstreams: 1 alinespagnolfedoce.pdf: 675452 bytes, checksum: 4344f40e93111c7814a7762ac2f2a71a (MD5) / Approved for entry into archive by Adriana Oliveira (adriana.oliveira@ufjf.edu.br) on 2016-10-05T11:33:55Z (GMT) No. of bitstreams: 1 alinespagnolfedoce.pdf: 675452 bytes, checksum: 4344f40e93111c7814a7762ac2f2a71a (MD5) / Made available in DSpace on 2016-10-05T11:33:55Z (GMT). No. of bitstreams: 1 alinespagnolfedoce.pdf: 675452 bytes, checksum: 4344f40e93111c7814a7762ac2f2a71a (MD5) Previous issue date: 2010-07-29 / CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / A síndrome metabólica (SM) é caracterizada por um grupo de distúrbios que predispõe a doenças cardiovasculares e ao diabetes mellitus tipo 2 (DM2). Obesidade abdominal, resistência à insulina, hipertensão, dislipidemia e um estado pró-inflamatório crônico são os principais componentes desta síndrome. Uma correlação bidirecional entre a SM e a periodontite tem sido sugerida, a partir do estado pró-inflamatório crônico. O objetivo deste estudo foi avaliar a influência da hiperglicemia no desenvolvimento da periodontite crônica e nos níveis séricos de interleucina-10, em indivíduos portadores de SM. Foram avaliados 30 indivíduos distribuídos em três grupos: NSM (não portadores de SM; n= 10); SMNG (portadores de SM normoglicêmicos; n= 10) e SMHG (portadores de SM hiperglicêmicos; n=10). A condição periodontal (profundidade de bolsa (PB), perda de inserção clínica (PIC), índice de placa (IP), sangramento a sondagem (SS) e número de dentes perdidos) e a condição metabólica (exames antropométricos e laboratoriais: glicemia em jejum (GJ), teste oral de intolerância a glicose (TOTG), colesterol HDL e triglicérides (TG)) foram avaliados em cada indivíduo. Os resultados demonstram que os indivíduos do grupo SMHG apresentam porcentagem de sítios com PIC ≥ 3 mm, PIC ≥ 5 mm; PB ≥ 3 mm, IP, SS e número de dentes ausentes mais elevados em relação aos indivíduos do grupo SMNG, esta diferença, porém, não foi estatisticamente significativa. No grupo SMHG, os indivíduos portadores de DM (n= 5) apresentaram porcentagem de sítios com PIC ≥ 5 mm estaticamente superior aos indivíduos pré diabéticos (p= 0,016). A porcentagem de sítios com PIC ≥ 5 mm foi correlacionada positivamente com os níveis de glicemia em jejum. Os indivíduos portadores de SM apresentaram níveis séricos de interleucina-10 mais elevados e diferiram estatisticamente dos indivíduos não portadores de SM (p= 0,003). O presente estudo sugere que a hiperglicemia influencia a gravidade da periodontite crônica e que os níveis aumentados de interleucina-10 estão relacionados com a SM, independente da condição glicêmica. / The metabolic syndrome (MS) is caracterized by a group of disorders that predisposes to cardiovascular diseases and to type 2 diabetes mellitus (T2DM). Abdominal obesity, insulin resistance, hypertension, dyslipidemia and a chronic proinflammatory state are the main components of this syndrome. A bidirectional correlation between MS and periodontitis has been suggested, from the chronic proinflammatory state. The aim of this study was to evaluate the influence of hyperglycemia on the development of chronic periodontitis and serum levels of interleukin-10 in individuals with MS. 30 individuals divided into three groups: NSM (non-MS patients, n = 10); SMNG (normoglycemic patients with MS, n = 10) and SMHG (hyperglycemic patients with MS, n = 10). The periodontal status (pocket depth (PD), clinical attachment loss (CAL), plaque index (PI), bleeding on probing (BOP) and number of missing teeth) and metabolic status (anthropometric and laboratory tests: fasting glucose (FG), oral test of impaired glucose tolerance (OGTT), HDL cholesterol and triglycerides (TG)) were evaluated in each individual. The results show that SMHG group subjects present higher percentage of sites with CAL ≥ 3 mm, CAL ≥ 5 mm, PD ≥ 3 mm, IP, SS and missing teeth than subjetcs in SMNG group, this difference however, was not statistically significant. In group SMHG, subjects with T2DM (n = 5) showed greater percentage of sites with CAL ≥ 5 mm than pre-diabetic subjects (p = 0.016). The percentage of sites with CAL ≥ 5 mm was positively correlated with levels of fasting blood glucose. Individuals with MS showed serum levels of interleukin-10 increased and differed significantly from the NMS group (p = 0.003). The present study suggests that hyperglycemia influences the severity of chronic periodontitis and that increased levels of interleukin-10 are related to the SM, independently on the glycemic status.

Made available in DSpace on 2014-08-20T14:38:00Z (GMT). No. of bitstreams: 1 dissertacao_anelize_felix.pdf: 541679 bytes, checksum: e35d84480d181fc2e13d21300abf6a98 (MD5) Previous issue date: 2010-02-24 / Demodicosis is considered one of the most severe canine skin disease. It is caused by the excessive proliferation of the Demodex canis mite, normal member of the canine skin. Skin lesions caused by the parasite predispose the skin to secondary infections that will further aggravate the patients clinical aspects. The objective of this work was to evaluate the clinical a histopathological evolution of the disease, as well as to study the seric levels of interleukin 10 (IL10) in demodicosis patients. The first study was conducted using 20 animals, 10 demodicosis patients (GD), and 10 control dogs (GC). All these animals were clinically evaluated, and submitted to deep skin scraping in search of Demodex mites. The dogs in the GD group were treated with moxidectin and evaluated in days 0, 20, 40, 60, and 80. Five of these animals were healed and submitted to skin biopsies on days 0 and 80, for the observation of histopathological alterations. A second study used 26 animals, 17 on G1 (demodicosis patients) and 9 on G2 (healthy dogs). All G1 animals were positive for demodicosis on the skin scrape test, and were submitted to clinical evaluation. Blood was harvested from all the animals, with the interleukin 10 dosage being carried out with the comrcial kit Quantikine Canine IL-10 Immunoassay® (R&D Systems) . Results obtained in the first experiment showed considerable clinical and scrape test improvement in GD, but no evolution in the histopathological patern. The GC presented higid skin and negative skin scrape test. In the second experiment, dogs in the G1 group presented hi clinical scores, indicating severe desiase. G1 had a mean IL10 level of 184,38 (+258,9) pg/mL, while the mean for G2 was11,94 (+ 2,27) pg/mL, indicating that hi levels of IL10 may be related to the development of the disease. The results described in this work show that, even when clinically healed, and with negative skin scrape test, demodicosis carriers present persistence of the lesions and the mite in the histological structure of the skin. This work also shows that demodicosis patients tend to have higher IL10 levels than healthy animals. / A demodicose é considerada uma das mais graves dermatopatias que acomete cães. É causada pela proliferação excessiva do ácaro Demodex canis, comensal da pele canina. Devido à lesão causada pelo parasita, a pele torna-se predisposta à instalação de infecções secundárias que agravam o quadro clínico do paciente. Objetivou - se avaliar a evolução clínica e histopatológica, assim como estudar os níveis de interleucina -10 sérica em cães portadores de demodicose. Para a primeira etapa, foram estudados 20 cães, 10 apresentando demodicose (GD) e 10 animais controle (GC). Todos foram avaliados clinicamente e submetidos a raspado cutâneo profundo para pesquisa de ácaros. Os animais do GD foram tratados com moxidectina e avaliados nos dias 0, 20, 40, 60 e 80. Cinco destes animais obtiveram cura e foram submetidos à biópsia cutânea no dia 0 e no dia 80, para análise das alterações histopatológicas. Na segunda etapa, foram utilizados 26 animais, 17 no G1 (portadores de demodicose) e 9 no G2 (cães hígidos). Todos os animais do GD tiveram raspado positivo para Demodex canis e foram avaliados clinicamente. Foi feita coleta de sangue para obtenção de soro em todos os animais sendo realizada dosagem dos níveis de interleucina 10 através do kit comercial Quantikine Canine IL-10 Immunoassay® (R&D Systems). Os resultados obtidos na primeira etapa, demonstraram uma melhora clinica considerável e negativação do raspado no GD, porém em relação ao padrão histopatológico não houve evolução. Quando comparados os dois aspectos, não houve diferença significativa. O GC apresentou pele hígida e raspado cutâneo negativo. Já na segunda etapa, os cães do G1 apresentaram escores clínicos altos, indicando severidade da doença. Os níveis de interleucina 10 no G1 tiveram média de 184,38 pg/ml (+258,9 pg/ml) enquanto o G2 apresentou média igual a 11,94 pg/ml (+ 2,27 pg/ml), indicando que níveis altos de IL10 podem estar relacionados com o desenvolvimento da doença. Os resultados demonstraram que mesmo clinicamente curados e com raspado cutâneo negativo, os cães portadores de demodicose apresentam persistência das lesões e do ácaro na estrutura histológica da pele, e também que cães com demodicose apresentaram níveis de interleucina 10 elevados quando comparados com animais sadios.

The most common human leukemia is B-cell chronic lymphocytic leukemia (B-CLL), which is characterized by a progressive accumulation of abnormal B-lymphocytes in blood, bone marrow and secondary lymphoid organs. Typically disease progression is slow, but as the number of leukemic cells increases, they interfere with the production of other important blood cells, causing the patients to be in an immunosuppressive state. To study the basis of this immunoregulation, we used cells from the transgenic Eμ-TCL1 mouse, which spontaneously develop B-CLL due to a B-cell specific expression of the oncogene, TCL1. Previously we showed that Eμ-TCL1 CLL cells constitutively produce an anti-inflammatory cytokine, IL-10. Here we studied the role of IL-10 in CLL cell survival in vitro and the development of CLL in vivo. We found that neutralization of IL-10 using anti-IL-10 antibodies or blocking the IL-10 receptor (IL-10R) using anti-IL-10R antibodies did not affect the survival of CLL cells in vitro. On the other hand, adoptively transferred Eμ-TCL1 cells grew at a slower rate in IL-10R KO mice vs. wild type (WT) mice. There was a significant reduction in CLL cell engraftment in the spleen, bone marrow, peritoneal cavity and liver of the IL-10R KO compared to WT mice. Further studies revealed that IL-10 could be playing a role in the tumor microenvironment possibly by affecting anti-tumor immunity. This was seen by a reduction in the activation of CD8+ T cells as well as a significantly lower production of IFN-γ by CD4+ T cells purified from CLL-injected WT mice compared to those purified from CLL-injected IL-10R KO mice. Also CLL-primed IL-10R null T cells were more effective than those from similarly CLL-primed wild type mice in controlling CLL growth in immunodeficient recipient mice. These studies demonstrate that CLL cells suppress host anti-tumor immunity via IL-10 production. This led us to investigate possible mechanisms by which IL-10 is produced. We found a novel role of B-cell receptor (BCR) signaling pathway in constitutive IL-10 secretion. Inhibition of Src or Syk family kinases reduces the constitutive IL-10 production by Eμ-TCL1 cells in a dose dependent manner. We identified the transcription factor Sp1 as a novel regulator of IL-10 production by CLL cells and that it is regulated by BCR signaling via the Syk/MAPK pathway.

Les monocytes jouent un rôle procoagulant majeur au cours du Syndrome Coronaire Aigu (SCA). Ils expriment à leur surface le Facteur Tissulaire (FT), initiateur majeur de la génération de la thrombine et sont à l’origine de la génération de icroparticules (MPs) procoagulantes exprimant également le FT. L’inhibition de la génération de thrombine et/ou de la génération des MPs monocytaires est ainsi d’un grand intérêt dans le contexte coronaire. Dans la première partie de ce travail, nous avons effectué une étude fondamentale évaluant, in vitro, l’effet d’anticoagulants utilisés dans le SCA, particulièrement le fondaparinux, ainsi que l’effet de l’interleukine-10 (IL-10), une cytokine anti-inflammatoire dotée des propriétés anti-athéromateuses. Ces deux types de molécules ont été analysées seules ou de façon combinée dans des modèles de génération de thrombine et de microvésiculation monocytaire. Ainsi, nous démontrons 1- que l’IL-10 inhibe lamicrovésiculation monocytaire, 2- que les molécules anticoagulantes inhibent d’avantage la génération de thrombine médiée par la MP monocytaire que celle médiée par le monocyte activé 3- que l’IL-10 potentialise l’effet anticoagulant du fondaparinux. Dans la mesure où la production de l’IL-10 est contrôlée génétiquement, nous avons évalué, dans la seconde partie de ce travail, l’association possible entre 5 polymorphismes génétiques de l’IL-10 et lerisque de pathologie coronaire aigue, et ce, dans une population tunisienne (291 patients/291témoins sains). Nous rapportons une association positive entre les variants polymorphes, IL-10 -592A (Odds ratio : 1,82) et IL-10R3 (Odds ratio : 1,46), et la pathologie coronaire. De façon intéressante, la littérature rapporte que ces deux variants polymorphes sont associés à une synthèse faible d’IL-10. En conclusion, nos résultats ne nous autorisent pas à proposer l’étude systématique des polymorphismes de l’IL-10 dans l’appréciation du risque coronaire aigu chez le patient. En revanche, nos résultats suggèrent que l’IL-10 pourrait constituer une molécule prometteuse dans l’arsenal des thérapeutiques anti-thrombotiques. / Procoagulant monocytes play a major role in the pathogeny of the acute coronary syndrome(ACS). Indeed, they express Tissue Factor (TF), the main trigger of thrombin generation, and generate highly procoagulant TF-bearing microparticles (MPs). It is therefore a major issue to control MPassociated thrombin generation in SCA. This led us to evaluate, in the first part of this work, the effect of anticoagulant molecules used in the management of ACS (including fondaparinux) but also of IL-10, an anti-inflammatory cytokine, which can modulate the progression of atheroma. Both types of molecules were evaluated separately or incombination, in a in vitro model of thrombin generation and monocytic MP generation. Ours results show that: 1- IL-10 inhibits monocytic-MP generation; 2- anticoagulants inhibit more potently MP-induced thrombin generation than activated monocyte-induced thrombin generation; 3- IL-10 potentiates fondaparinux inhibitory effect on thrombin generation.As IL-10 production is genetically controlled, we evaluated the possible influence of 5 well-described IL- 10 polymorphisms on the risk of ACS among Tunisians (291 patients/291 healthy controls). Results show that two polymorphic variants of IL-10 i.e. SNP-592A (Odds ratio: 1,82) and microsatellite IL-10R3 Odds ratio: 1,46) are significantly associated with the risk of SCA. Interestingly, the literature reports that these two polymorphic variants are associated with low levels of IL-10 production. In conclusion: Our results do not allow us to recommend the analysis of IL-10 polymorphisms in the assessment of ACS risk. However, our data suggest that IL-10 is a promising antithrombotic pharmacological agent, in this clinical situation.

Orientador: Márcio Zaffalon Casati / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Odontologia de Piracicaba / Made available in DSpace on 2018-08-27T08:08:59Z (GMT). No. of bitstreams: 1 Monteiro_MabelledeFreitas_M.pdf: 1431989 bytes, checksum: 55bf42377e6ffeb3626cb5a8d6551c1c (MD5) Previous issue date: 2015 / Resumo: O Resumo poderá ser visualizado no texto completo da tese digital / Abstract: The Abstract is available with the full electronic digital document. / Mestrado / Periodontia / Mestra em Clínica Odontológica

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