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Avaliação imunológica de idosos no pré e pós-operatório de correção de valvulopatia cardíaca / Immune evalution of elderly subjects submitted to valvulopathy correction surgeryIrina Ewers 19 March 2009 (has links)
Sabe-se que o sistema imune, através de um fenômeno denominado imunossenescência, gradativamente diminui a sua capacidade de resposta durante a vida. Este fato pode tornar o indivíduo mais suscetível a infecções e outras patologias. Neste contexto, seria útil procurar por fatores que alterassem esta evolução natural, principalmente os capazes de acelerar este processo. Por esta razão, nós procuramos por diferenças nos parâmetros imunológicos entre o antes e o depois da cirurgia de valva cardíaca em idosos com mais de 65 anos. Nossos resultados não apontaram, no pós-operatório, para uma diminuição da capacidade imune, uma vez que os testes cutâneos de hipersensibilidade para o PPD, tricofitina e candidina não se alteraram. Quando a resposta linfoproliferativa foi avaliada in vitro, também não apresentou diferença. Por outro lado, nós observamos um aumento na porcentagem de células T CD3 +, T CD4 + e monócitos no sangue periférico, quando comparamos os períodos. Sendo que os marcadores de ativação cellular CD25 +, CD69 + e o CD95 também se apresentaram elevados. Quanto a secreção de citocinas, nossos resultados apontam para um amento de IL-4 e IL-8. Inversamente, concentrações reduzidas de IL-2, IL-12 e IFN- foram detectadas no sobrenadante de PBMCs quando estimuladas in vitro. Em suma, nossos dados demonstram que a cirurgia de valva cardiaca é capaz de alterar vários parâmetros da resposta immune, com um aumenrto da porcentagem de células, quanto da expressão de marcadores de ativação celular e secreção de citocinas / It is known that the immune system, through a phenomenon called immunosenescence, undergoes functional changes during life which may culminate in a diminished capacity of response, turning the subject more susceptible to infections and other pathologies. In this context, it is useful to search for factors that alter this natural evolution, mainly able to delay this process. For this reason, we assessed different immunologic parameters before and after cardiac valve surgery in 65 year-old patients. Our results did not point to a postoperative immunedeficiency-like state, once that the cutaneous tests to PPD, candidin and tricophytin remained positive for most of the subjects. When the proliferative response was assessed in vitro, there were also no differences. On the other hand, we observed a post-surgical increase in the percentage of T CD3 +, T CD4 + cells and in monocytes from peripheral blood when we compare both periods. Moreover, it is important to highlight that activation markers, such as CD25, CD69 and CD95 were also presented in higher levels. According to the cytokine secretion, our results appointed to a greater secretion of IL-4 and IL-8 postoperative. Conversely, reduced concentrations of IL-2, IL-12 and IFN- were detected in supernatant of PBMCs when stimulated in vitro. In summary, our data reveal that the cardiac valve surgery with extra corporeal procedure and anesthesia is able to alter several parameters of the immune response, with an increased percentage of the major assessed cells, as well as in the expression of activation markers and cytokine secretion
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Respostas comportamentais e metabólicas à ativação do sistema imune por injeção de lipopolissacarídeo (LPS) em Scinax gr. perpusillus (Anura: Hylidae) / Behavioral and metabolic responses to immune system activation by lipopolysaccharide (LPS) in Scinax gr. perpusillus (Anura: Hylidae)Letícia Regina do Amaral Braga 26 November 2013 (has links)
Parasitas e agentes infecciosos devem representar uma importante pressão seletiva atuante sobre a evolução de aspectos da história de vida dos hospedeiros. Os desafios impostos aos hospedeiros pelos patógenos resultam em uma séria de respostas fisiológicas e comportamentais, respostas estas pouco conhecidas para a maioria das espécies de hospedeiros ectotermos, como anfíbios anuros. O objetivo deste estudo foi quantificar os efeitos da ativação do sistema imune via injeção de LPS (lipopolissacarídeo), em duas diferentes concentrações, sobre o comportamento alimentar, taxa metabólica de repouso, atividade locomotora espontânea e comportamento anti-predatório de machos adultos da perereca das bromélias: Scinax gr. perpusillus . O tratamento com LPS provocou redução na taxa de alimentos ingeridos, confirmando o efeito anorexigênico deste componente, e aumento na atividade locomotora espontânea, porém não apresentou efeito sobre a taxa metabólica de repouso ou sobre o comportamento anti-predatório das pererecas / Parasites and infectious agents should represent important selective pressure acting on the evolution of aspects of life history of the hosts. The challenges posed by pathogens to hosts result in a series of physiological and behavioral responses, which are not well known to most ecthoterm hosts species, such as anuran amphibians. The aim of this study was to quantify the effects of activation of the immune system via injection of LPS (lipopolysaccharide) in two different concentrations, on feeding behavior, resting metabolic rates, spontaneous locomotor activity, and antipredatory behavior of adult males of the bromeliad tree-frog: Scinax gr. perpusillus . Treatment with LPS caused a reduction in the rate of food intake confirming the anorexigenic effect of this component and an increase in spontaneous locomotor activity, but had no effect on the resting metabolic rate or in the the antipredator behavior of frogs
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O papel modulador do gene Aire (autoimmune regulator) sobre redes de expressão gênica em células tímicas epiteliais medulares / Promiscuous gene expression in medullary thymic epithelial cells is connected in network where the Aire gene is an upstream modulatorClaudia Macedo 28 March 2008 (has links)
A expressão de antígenos restritos a tecidos (TRAs do inglês tissue restricted antigens) no timo pelas células epiteliais medulares (mTECs de medullary thymic epithelial cells) é essencial para a tolerância central das células T. Devido à sua heterogeneidade em termos de representação de autoantígenos, esse fenômeno foi denominado como expressão gênica promíscua (PGE de promiscuous gene expression), no qual o gene Aire (de autoimmune regulator) desempenha um papel como principal regulador transcricional positivo sobre um grande conjunto de TRAs dependentes de Aire. A proteína Aire tem a capacidade de interagir com seqüências específicas de DNA desempenhando um papel como regulador direto. Neste estudo utilizamos o método dos cDNA microarrays para acessar a PGE em células mTEC CD80+ murinas cultivadas in vitro. O agrupamento hierárquico dos dados permitiu a observação de que os genes de TRAs foram diferencialmente expressos. Para testar essa hipótese, inicialmente silenciamos o gene Aire pelo método de RNA interferente (RNAi) nas células mTEC. O agrupamento hierárquico dos dados de cDNA microarray mostrou um conjunto de genes de TRAs dependentes de Aire, os quais foram reprimidos após o silenciamento deste último. Redes gênicas reconstruídas desses dados permitiram a identificação de um nó gênico (Gucy2d) estabelecendo regulação positiva sobre genes downstream nas células mTEC normais. Entretanto, sob efeito do silenciamento de Aire, Gucy2d passou a ser um repressor. Esses resultados evidenciaram que genes da PGE estão conectados em rede, que um nó gênico pode atuar como intermediário no seu controle e que Aire na rede PGE desempenha seu controle como regulador upstream. / The expression of tissue restricted antigens (TRAs) in thymus by medullary thymic epithelial cells (mTECs) is essential for the central selftolerance of T cells. Due to heterogeneity of autoantigen representation this phenomenon has been termed promiscuous gene expression (PGE), in which the autoimmune regulator (Aire) gene plays a role as main positive transcriptional regulator on a large set of Aire-dependent TRAs. Aire protein is able in binding to specific DNA sequence motifs and plays a role as a direct regulator. Here we used the cDNA microarray method to access PGE in murine CD80+ mTECs cultured in vitro. Hierarchical clustering of the data allowed observation that TRA genes were differentially expressed. To further investigate the control of PGE, we hypothesize that TRA genes establish networks contributing it selves to modulate their transcriptional levels. Aire in this case plays a role as upstream positive modulator. To test this hypothesis, initially we silenced Aire by gene knockdown (RNA interference) in mTECs. Hierarchical clustering of cDNA microarray data showed a set of Airedependent TRAs genes, which were down regulated after Aire silencing. Gene networks reconstructed from these data allowed the identification of a gene node (Gucy2d) establishing positive regulation upon downstream genes in normal mTECs. Nevertheless, under silencing of Aire, Gucy2d has become a repressor. These finding evidentiate that, genes features in PGE are connected in network; a gene node may act as intermediate in their control and that Aire in PGE network plays a role as an upstream regulator.
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Avaliação da expressão de indoleamina 2, 3 dioxigenase - IDO nos leucócitos presentes no tumor ascítico de Ehrlich perante o bloqueio da via de ativação linfocitária B7+CTLA-4 / Influence of the lymphocyte activation pathway B7 + CTLA-4 on the expression of indoleamina 2, 3 dioxygenase - IDO in leucocytes present in Ehrlich ascites tumorCarla Simone Soares 02 August 2017 (has links)
O tumor de Ehrlich (TE) foi descrito inicialmente, como adenocarcinoma mamário de camundongos, desenvolvendo-se na forma ascítica ou sólida a partir de sucessivos transplantes no peritônio ou tecido subcutâneo destes animais. O tumor ascítico de Ehrlich (TAE) tem sido utilizado como um tumor transplantável para o desenvolvimento de pesquisas relacionadas à oncologia. Estudos tem demonstrado que o desenvolvimento de TAE resulta no estímulo de células citotóxicas, como os linfócitos T e células Natural Killer (NK), mediadas principalmente por macrófagos. Os macrófagos e as células dendríticas (DCs), podem induzir a síntese da enzima indoleamina 2,3 dioxigenase (IDO) em tecidos tumorais, via ligação das moléculas co-estimuladoras B7-1 e B7-2, presentes nestas células, com a molécula CTLA-4 (antígeno 4 associado a linfócitos T citotóxicos), presente em linfócitos T reguladores CD4+ CD25+. A IDO é uma enzima que degrada o aminoácido essencial triptofano, processo que além de levar à privação do mesmo no microambiente celular, gera metabólitos que impedem a ativação e proliferação de linfócitos T e, consequentemente, mecanismos a eles associados como os de rejeição podem ser seriamente comprometidos. Mediante o exposto e pela presença de células imunológicas que expressam IDO no microambiente do tumor ascítico de Ehrlich, este trabalho teve como objetivo verificar a expressão da IDO após o bloqueio da interação B7/CTLA-4 por meio da citometria de fluxo. De acordo com as análises realizadas, os resultados demostraram que houve redução de 4,9% para 2,53% na expressão da enzima IDO. Em face dos resultados, parece plausível sugerir que o bloqueio desta via de ligação foi eficaz na redução dos níveis de atividade da IDO, o que poderia restaurar a capacidade de resposta dos linfócitos T contra as células tumorais. Nesta perspectiva sobre a IDO como mediadora no controle do escape imune feito pelas células tumorais, tais resultados podem colaborar para modulação desta enzima no microambiente tumoral. / The Ehrlich tumor (TE) was first described, as breast adenocarcinoma of mice. The TE develops in ascitic or solid form from successive transplantations in the peritoneum or subcutaneous tissue of these animals. The Ehrlich ascites tumor (TAE) has been used as a transplantable tumor for the development of research related to oncology. Studies have shown that the development of TAE results in stimulation of cytotoxic cells, such as T lymphocytes and Natural Killer cells (NK), mediated mainly by macrophages. Macrophages and dendritic cells (DCs) in tumor tissues, via co-stimulation of molecules B7-1 and B7-2, present in these cells, toghether with the CTLA-4 (Cytotoxic T-Lymphocyte-Associated antigen 4) molecule, present on regulatory T CD4 + CD25 + lymphocytes , may induce the synthesis of indoleamine 2, 3 dioxygenase (IDO). IDO is an enzyme that catabolizes the essential amino acid tryptophan, impairing activation and proliferation of T lymphocytes and consequently, compromising mechanisms associated with them such as rejection. Considering the presence of immune cells in the tumor microenvironment Ehrlich ascites that express IDO, this study aimed to verify the expression of IDO after the blockade of interaction B7/CTLA-4 by flow cytometry. Results demonstrated that there was a reduction of 4.9% to 2.53%in the expression of IDO. Given the results, it seems plausible to suggest that blocking this binding via was effective in reducing the levels of expression of IDO, which could restore the responsiveness of T cells against tumor cells. In this perspective on the IDO as a mediator in the control of immune escape made by tumor cells, these results may collaborate for modulation of this enzyme in the microenvironment.
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Activation of TLR5 by Salmonella-derived flagellinsMetcalfe, Hannah Jane January 2012 (has links)
No description available.
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The Effects of Long Term Moderate Ethanol Intake on the Immune Response in RatsFollin, Cynthia A. (Cynthia Ann) 08 1900 (has links)
Using a rat model, the effects of a single dose or six to twelve months of daily oral administration of ethanol on the immune system were determined. The rats were challenged with sheep red blood cells after the various dosing periods to elicit an immune response. Immune system responsiveness was determined by means of white blood cell counts and differentials, antibody titers, and T-cell numbers. No deleterious effects of the ethanol on the immune response were seen, while the female alcohol-fed rats showed a significant increase in T-Cell numbers, white blood cell counts, and lymphocytes over the sham group.
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Psychological, Social, and Immunological Outcomes following Marital SeparationHasselmo, Karen Elizabeth, Hasselmo, Karen Elizabeth January 2017 (has links)
Close relationships play an integral role in human health (Coan & Sbarra, 2015). It follows, then, that the dissolution of an important relationship can have a variety of negative health consequences (Amato, 2010; Kitson & Morgan, 1990; Sbarra, Law, & Portley, 2011), and social loss confers vulnerability to a range of morbidities and early mortality. Disrupted marital status is one of the strongest sociodemographic predictors of stress-induced physical illness (Somers, 1979) and marital disruption has long been reported as one of life's most stressful events (Bloom, Asher, & White, 1978). Robust evidence links divorce or separation to poorer health outcomes; however, the exact mechanisms through which relationship dissolution influences our health so profoundly are not yet fully elucidated (Sbarra, Hasselmo, & Bourassa, 2015). The current study investigated how changes in psychological responses to divorce are associated with changes in immune responding in recently-separated adults (N = 55). I followed participants over an average of five months, collecting psychological distress measures at three visits, each one month apart, and immune measures at two visits, five months apart. To assess how variability in social engagement is associated with immunological responses following the end of a marriage, I incorporated naturalistic, observational data using a new methodology. I found that an objectively derived composite of social behaviors including (a) time spent with others; (b) time spent socializing/entertaining; (c) time spent in substantive conversation; and (d) time spent receiving positive support predicted concurrent immune outcomes over and above the effects of psychological distress and/or loneliness, and that psychological distress may exert indirect influence on immune outcomes through social integration. Furthermore, attachment style revealed differential longitudinal associations between social integration and immune outcomes. This research expands current knowledge on the immune-relevant outcomes of divorce and separation, and includes new methodology for naturalistically-derived measures of social engagement in determining how this common life stressor is associated with health over time.
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Evaluation of botanical extracts with immune enhancing and /or anti-HIV activity in vitroBrink, Mnandi 10 November 2011 (has links)
M.Sc. / To successfully intervene in the HIV/AIDS pandemic, knowledge of the pathogenesis of the disease and factors that stimulate or inhibit viral replication are crucial. Plants are expected to produce antiviral compounds since viruses form one of the major groups of plant pathogens. The objective in this project was to investigate the effects of 6 plant extracts on immune responses as well as evaluate their potential anti-HIV activity. Plant species tested were: Hypoxis hemerocal/idea, Elephantorrhiza elephantina, Spirulina platensis, Echinacea purpurea, Echinacea pal/ida and Cannabis sativa. Extracts were prepared via 24 hour extraction or 12 hour reflux in H20, methanol and ethanol in a 1:5 ratio. The crude extracts were analysed by TLC and HPLC and shown to consist of complex related mixtures of compounds. Using LC-MS, partial identification of methanol extracts revealed the following expected compounds: 9- octadecenoic acid (E)- in Hypoxis hemerocallidea, 4H-1-benzopyran-4-one,2-(3,4- dihydroxyphenyl)-7 -(13-D-glucopyranosyloxy)-5-hydroxy- in E/ephantorrhiza e/ephantina, ethanol,2-butoxy-,phosphate in Spirulina platensis, 2-propenoic acid,3-(3,4-dihydroxyphenyl)- in Echinacea purpurea, 2-propenoic acid,3-(3,4-dihydroxyphenyl)- in Echinacea pal/ida and ~-9- tetrahydrocannabivarin in Cannabis sativa tincture. Viability assays using tetrazolium salts (XTT) gave a qualitative picture of events allowing us to assess host cell responses and extract toxicity. Extracts exhibited intrinsic absorbances at some visible wavelengths but did not interfere at the wavelengths used in this viability assay. Having analysed cell viability it was thought prudent to report on the type of cell death induced by either HIV or the extracts, so Annexin-V (indicator of apoptosis) and PI (indicator of necrosis) detected by flow cytometry was employed. Results obtained revealed that cells were driven towards necrosis rather than apoptosis. None of the extracts showed significant in vitro toxicity in CEMss, CEMNKR. U937, Jurkat and PM1 cells or ex vivo in PBMC at a concentration range of 1000f..lg/ml-4f.!g/ml. Viability assays were also an indirect indication of HIV's effect on the cells. As for the effect of extracts on the immune system, IL-2 secretion was stimulated by most of the extracts. The effect of plant extracts on HIV activity was also investigated by looking at core protein levels (p24 was generally decreased by methanol extracts), reverse transcriptase activity (no detectable influence) and envelope glycoprotein levels (gp120 levels were only marginally reduced). It appears that Echinacea purpurea, Echinacea pal/ida and Spirulina platensis have immune enhancing abilities, while Hypoxis hemerocallidea, Elephantorrhiza e/ephantina and Cannabis sativa have dual purposes by enhancing both immunity and inhibiting HIV activity.
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Multiple epitope immunogens (MEI) mimic the variability of the V3 loop of HIV-1 subtype C.Hewer, Raymond 09 May 2008 (has links)
M.Sc. / Therapeutic and preventative treatment are continually being sought to cease or curtail the worldwide HIV-1 epidemic. At present, therapeutic drug strategies such as highly active anti-retroviral therapy (HAART) have been particularly successful in slowing disease progression and reducing the incidence of AIDS and AIDS related mortality (Detels et al., 1998; Mocroft et al., 1998; Palella et al., 1998). However, the high costs, intricate dosing regimens and limited availability of the HAART drugs (Butera, 2000) has restricted its efficacy in developing and third world countries. As such, available and future drugs will remain inaccessible to the regions that are profoundly affected by the epidemic. An effective vaccine presents a viable solution to the HIV-1 epidemic in these countries. Approximately 70 vaccines are presently in various stages of clinical trials, the majority of which are subtype B specific (Johnston and Flores, 2001). This prevents their use in the predominantly subtype C infected sub-Saharan region of Africa, which accounts for 50% of the global HIV / AIDS population and includes South Africa, statistically the country with the highest number of people living with HIV / AIDS of any country in the world (UNAIDS, 2002). Presently there is no HIV-1 vaccine, regardless of subtype, in clinical use. This owes to several difficulties that hinder the progression of vaccine development, including the lack of predictive animal models, the establishment of viral latency and the difficulty involved in overcoming HIV-1 genetic diversity (Klein, 1999). The expansive HIV-1 genetic variation exhibited by HIV-1 is attributed to a high number of errors made by the reverse transcriptase (RT) enzyme (Coffin, 1992) and the absence of RT proofreading mechanisms during HIV-1 replication (Roberts et al., 1988; Bebenek et al., 1989). The HIV-1 nucleotide sequence drift is most frequently observed in the envelope (env) gene and expressed in env gene products (Shafer et al., 1999). Expression of the variable genome results in the production of progeny strains that are not identical to the parent strain (i.e. HIV-1 exists as a quasispecies within each seropositive individual and between individuals) and contributes to the diverse collection of viral strains in global circulation that vary across and within subtypes. Thus, for an HIV-1 vaccine to be efficient and truly functional it would be required to target this observed hypervariability and be effective against a multitude of currently circulating strains, exhibit cross-clade specificity and remain viable despite the emergence of variant strains. In this study we describe the design, synthesis and immunological ability of a multiple epitope immunogen (MEI) that mimics the hypervariability observed within the third variable (V3) loop of the envelope gp120 region of HIV-1 subtype C. Conjugation to a multiple antigenic peptide (MAP) produces a four -branched (b4) tetrameric peptide construct, designated MEIV3b4. This construct was characterized by theoretical and analytical techniques, tested in a variety of immunological assays and assessed for its potential as a candidate vaccine component. The construct was comparatively analysed through evaluation of three comparison peptides, two of which are hypervariable and based on the V3 region, the other representing a conserved region of HIV-1 envelope. The V3 peptides, named b-MEI-s and poly-L-MEI, differ from the MEIV3b4 construct in that they are less variable and less branched or conjugated to a traditional carrier rather than to a MAP system, respectively. The conserved peptide, designated CCD4 allowed for comparative evaluation between conserved and variable peptides as potential vaccine components. / Dr. Debra Meyer
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Proteomic analysis of the humoral antifungal immune response of the soft tick,Ornithodoros savignyi Audouin (1827)Stopforth, Elaine 18 February 2010 (has links)
Ticks are blood feeding ectoparasites that ingest large volumes of vertebrate blood. They are the most important arthropods that are capable of transmitting pathogens which cause disease in humans and domestic animals. Ticks are exposed to various microorganisms during feeding as well as in their habitat. They therefore must have a very good immune system to recognize and destroy these microorganisms. In the present study a micro-broth dilution assay was used to determine whether antifungal activity was present in different tick tissue extracts with or without challenge. The midguts gave the highest inhibition of yeast growth, followed by the salivary glands and then the hemolymph. This was seen with unchallenged tick tissue extracts, as well as tissue extracts collected after yeast challenge (2 hours). Thus all of the tick tissue extracts that was analyzed in this study had antifungal activity. Proteomics was used to determine whether proteins were differentially expressed in the hemolymph plasma, after a fungal challenge. 2DE was used since proteins are not only separated by molecular mass, but also by their charge. The proteins that were separated on the 2D-gels ranged between 17.5-76 kDa and not all proteins present on the 1D-gels (14-97 kDa) could be seen on the 2D-gels. Ticks were challenged for 2 hours to define the proteins that play a role in the short term innate immune response during a fungal infection. Various proteins were differentially expressed in the hemolymph samples that were collected 2 hours after ticks were injected with saline, â-1,3-glucan or yeast (or 72 hours). Injury and fungal challenge play a role in producing proteins that might play a role in the fungal response of the tick. Five spots that were statistically significant in the hemolymph collected 2 hours after ticks were injected with yeast cells were analyzed with MS/MS. No matches were found with MASCOT database searching or with EST searching. This can be due to the limited information that is available on the soft ticks, as only hard tick ESTs heve been published. It was also attempted to identify hemolymph proteins that might play a role in the recognition of fungi. Hemolymph was incubated with live Candida albicans cells and eluted with buffer. Three protein bands (97, 88 and 26 kDa) were found to be present whether ticks were challenged or unchallenged. These proteins were subjected to MS/MS analysis and database searching was performed revealing no matches to other known proteins. The antifungal response was found to be present in the soft tick O. savignyi and might play a vital function in the innate immune response during a fungal infection. These proteins may serve as lead molecules that could be used in the development of novel antifungal drugs, as well as in vaccine development. Copyright / Dissertation (MSc)--University of Pretoria, 2010. / Biochemistry / unrestricted
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