Thesis (M.S.)--Youngstown State University, 2008. / Includes bibliographical references (leaves 73-74). Also available via the World Wide Web in PDF format.

Thesis (Ph. D.)--University of Hong Kong, 2008. / Also available online.

Thesis (Ph. D.)--University of Hong Kong, 2008. / Also available in print.

Thesis (doctoral)--Uppsala University, 1983. / Includes bibliographical references (p. 31-35).

Thesis (Ph. D.)--Ohio State University, 2006. / Title from first page of PDF file. Includes bibliographical references (p. 185-219).

Ο ρόλος της φαγοκυτταρικής ικανότητας των μονοκυττάρων και των πολυμορφοπυρήνων στην σήψη δεν έχει μελετηθεί επαρκώς. Η παρούσα έρευνα είχε ως σκοπό να εκτιμήσει την επίδραση της φαγοκυτταρικής δραστηριότητας των μακροφάγων στην τελική έκβαση ασθενών με βαριά σήψη. 31 ασθενείς και 30 υγιή άτομα πήραν μέρος στην μελέτη. Η φαγοκυττάρωση των πολυμορφοπυρήνων και των μονοκυττάρων εκτιμήθηκε τις πρώτες 24 ώρες μετά την εισαγωγή του ασθενούς ενώ τα αποτελέσματα συσχετίσθηκαν με την έκφραση του CD64 στα πολυμορφοπύρηνα και στα μονοκύτταρα, την έκφραση του CD14 στα μονοκύτταρα, το SAPSII score και την επιβίωση των ασθενών. Η ελαττωμένη φαγοκυττάρωση των ουδετεροφίλων τις πρώτες 24 ώρες μετά την εισαγωγή ήταν αρνητικός προγνωστικός δείκτης για την επιβίωση. Η αυξημένη έκφραση τoυ CD64 τόσο στα PMN όσο και στα μονοκύτταρα επηρέασε θετικά την πρόγνωση των ασθενών. Σε πολυπαραγοντική ανάλυση η φαγοκυτταρική δραστηριότητα των πολυμορφοπυρήνων ήταν ο μοναδικός ανεξάρτητος προγνωστικός παράγοντας για τη επιβίωση. Ασθενείς με φαγοκυτταρική δραστηριότητα PMN<37% παρουσίασαν μικρότερη έκφραση του CD64 στα πολυμορφοπύρηνα και στα μονοκύτταρα και κακή πρόγνωση ενώ ασθενείς με φαγοκυτταρική δραστηριότητα PMN>37% παρουσίασαν μεγαλύτερη έκφραση CD64 στα ουδετερόφιλα και στα μονοκύτταρα και καλή πρόγνωση. Η ελαττωμένη φαγοκυτταρική ικανότητα των ουδετεροφίλων πιθανά αντιπροσωπεύει μία κατάσταση ελαττωμένης δραστηριότητας παρόμοια με εκείνη των μονοκυττάρων κατα την διάρκεια του CARS (Compensatory Anti-inflammatory response syndrome, σύνδρομο αντισταθμιστικής αντιφλεγμονώδους απάντησης). Η χημειοτακτική δραστηριότητα των πολυμορφοπυρήνων στους επιβιώσαντες ασθενείς ήταν σημαντικά αυξημένη σε σχέση με τους ασθενείς που απεβίωσαν ενώ αντίθετα δεν παρουσιάσθηκε στατιστικά σημαντική διαφορά στην χημειοτακτική ικανότητα των ουδετεροφίλων ανάμεσα στην ημέρα της εισαγωγής και την ημέρα του εξιτηρίου. H χορήγηση του αυξητικού παράγοντα G-CSF στους ογκολογικούς ασθενείς με εμπύρετο ουδετεροπενία δεν επηρέασε την φαγοκυτταρική δραστηριότητα των πολυμορφοπυρήνων σε σημαντικό βαθμό. / The role of the effector function of monocytes and neutrophils in sepsis has been poorly investigated. The present study assessed the phagocytic activity of monocytes and neutrophils and evaluated its predictive significance in septic patients 31 patients with severe sepsis and 30 healthy individuals were enrolled in the study. The phagocytic activity of monocytes and neutrophils was evaluated and the results were correlated to the expression of CD64 on neutrophils and monocytes, CD14 antigen on monocytes, the SAPS score and the patients’ survival. The phagocytic activity of polymorphonuclears (PMN) in 24 hours after admission was decreased in all patients. Patients with PMN phagocytic activity <40% had lower expression of CD64 on monocytes and PMN and worse outcome while those with phagocytic activity>40% had higher expression of CD64 on monocytes and PMN and better outcome. In multivariate analysis the phagocytic activity of PMN was the only independent predictor factor for patients’ survival. The phagocytic activity of neutrophils in septic patients is a significant parameter of the final outcome. The upregulation of PMN CD64 expression is prerequisite for their increased phagocytic function but does not reflect it. The chemotactic functionof neutrophils was significantly increased in survivors compared to non survivors.In contrast, no statistical significance of chemotactic activity of PMNs was detected between the admission day and the day of discharge. The administration Of G-CSF in cancer patients with febrile neutropenia did not increase the phagocytic activity of neutrophils.

Made available in DSpace on 2015-12-06T23:45:12Z (GMT). No. of bitstreams: 0 Previous issue date: 2008 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / Paracoccidioidomicose (PCM) é uma doença granulomatosa sistêmica causada pelo fungo dimórfico térmico Paracoccidioides brasiliensis. O principal antígeno desse fungo é uma glicoproteína de 43-KDa. Gp43 possui diferentes funções: participa do mecanismo de evasão durante a instalação da infecção primária e estimula formação de granuloma in vitro, apresentando epítopos reconhecidos por células T que induzem resposta protetora contra o P. brasiliensis. Aqui, investigamos quais epítopos da gp43 poderiam diminuir a função fagocítica de macrófagos e a reação inflamatória correspondente. Diferentes peptídeos da gp43 foram adicionados a culturas de macrófagos derivados de medula óssea. Posteriormente, foram desafiados com zymosan e os índices fagocíticos foram determinados. A expressão dos peptídeos na superfície da molécula foi determinada por análises gráficas usando o módulo Protean; DNAstar Inc. Dois peptídeos que diminuíram índices fagocíticos e estavam expressos na superfície da gp43, P4 e P23, foram selecionados para ensaios posteriores. Mostramos que ambos, P4 e P23 inibiram a liberação de NO por macrófagos estimulados com zymosan enquanto aumentaram liberação de H2O2. A liberação de TNF-α no sobrenadante de cultura de testes fagocíticos in vitro também foi inibida na presença dos peptídeos. Ensaios in vivo com Mycobacterium bovis - bacillus Calmette-Guérin (BCG) demonstraram que estes peptídeos também apresentaram propriedades antiinflamatórias não-específicas a PCM. / Paracoccidioidomycosis (PCM) is a systemic granulomatous disease caused by Paracoccidioides brasiliensis, a thermal dimorphic fungus. Its major antigen is a 43-kDa glycoprotein. Gp43 embodies different functions: it participates in the evasion mechanisms during the installation of primary infection and stimulates granuloma-like formation in vitro, presenting T-cell epitopes that induce protective response against the fungus. Here, we investigated which epitopes from gp43 could inhibit both, macrophage functions and inflammatory reaction. Different gp43 peptides, spanning the entire sequence of the molecule, were added to cultures of bone marrow-derived macrophages. After challenge with zymosan, phagocytic indexes were measured. Peptides expressed on the molecule surface were determined by graphic analysis using the Protean module; DNAstar Inc. Two peptides which decreased phagocytic index and were expressed in surface of molecule, P4 e P23, were selected for further studies. It was shown that both, P4 e P23 inhibited the release of NO by zymosan stimulated macrophages while enhancing release of H2O2. The release of TNF-α in culture supernatants from in vitro phagocytic tests was also inhibited in their presence. In vivo assays with Mycobacterium bovis - bacillus Calmette-Guérin (BCG) demonstrated that these peptides also presented non-specific anti-inflammatory property. / BV UNIFESP: Teses e dissertações

Made available in DSpace on 2015-12-10T14:22:52Z (GMT). No. of bitstreams: 0 Previous issue date: 2015-05-15. Added 1 bitstream(s) on 2015-12-10T14:29:05Z : No. of bitstreams: 1 000851662_20160515.pdf: 427931 bytes, checksum: 4ef8d142c5c5234191158ab7c2efea52 (MD5) Bitstreams deleted on 2016-05-16T14:18:42Z: 000851662_20160515.pdf,. Added 1 bitstream(s) on 2016-05-16T14:19:23Z : No. of bitstreams: 1 000851662.pdf: 1696866 bytes, checksum: d81719395199b25212968ab82e23a88e (MD5) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) / A fagocitose de células apoptóticas é um processo dinâmico importante para a homeostase dos tecidos após injúria. Os macrófagos além de atuarem na remoção de células apoptóticas, também colaboram na defesa contra microrganismos. Existem ao menos duas populações de macrófagos classificadas como macrófagos M1 (pró-inflamatórios) e macrófagos M2 (anti-inflamatórios). Estes leucócitos diferem quanto ao fenótipo e funções efetoras dependendo, primordialmente, do microambiente e dos microrganismos com que interagem no tecido. Em uma situação de injúria pulmonar estéril (inalação de agentes tóxicos), há acúmulo de células apoptóticas sem a presença de agente microbiano. Por outro lado, durante uma infecção pulmonar bacteriana, ocorre intensa migração de células para o local da infecção na tentativa de conter a proliferação bacteriana, resultando em intenso acúmulo de células apoptóticas infectadas neste tecido. Portanto, nesse trabalho foi avaliado, in vitro e in vivo, o efeito da fagocitose de células apoptóticas infectadas (AC-Sp) ou estéreis (AC) na polarização de macrófagos M1/M2, bem como suas funções efetoras, e o efeito da PGE2 nesse contexto de eferocitose. Macrófagos M0 co-cultivados com AC ou AC-Sp apresentam um fenótipo intermediário entre M1 e M2, com secreção de altos níveis de IL-6, TNF-α, PGE2, TGF-β e NO e a expressão de genes relacionados ao perfil M1, iNOS, e ao perfil M2, Arginase 1. Além disso, macrófagos M0 cultivados com AC ou AC-Sp, apresentam supressão da função microbicida quando desafiados com S. pneumoniae. A presença de PGE2 dirige a polarização de macrófagos M0 a um perfil M2 e a presença de inibidores de COX promove a reversão do fenótipo de polarização destes macrófagos. Os resultados in vivo demonstram que a instilação de AC ou AC-Sp promove distintos microambientes no pulmão destes animais, que influenciam na resolução da infecção..... / The phagocytosis of apoptotic cells is dynamic and crucial for homeostasis after injury. Macrophages act on the clearance of apoptotic cells and collaborate with defense against microorganisms. There are at least two distinct macrophage populations classified as M1 macrophages (pro-inflammatory) and M2 macrophages (anti-inflammatory). Both cells differ on the state of polarization and effectors function depending primarily on the microenvironment and microorganisms that interact into the tissue. In a situation of sterile lung injury (inhalation of toxic agents), there are accumulation of apoptotic cells without the presence of pathogen. On the other hand, during a bacterial pulmonary infection there is intense cell migration to the infection site in an attempt to impair bacterial growth, resulting in a large accumulation of infected-apoptotic cells in the tissue. Therefore, this study has evaluated in vitro and in vivo, the effect of the phagocytosis of infected-apoptotic cells (AC-Sp) or sterile cells (AC) on the polarization of M1/M2 macrophages, as well as in their effector functions and the effect of PGE2 in the context of efferocytosis. M0 macrophages co-cultured with AC or AC-Sp show an intermediate phenotype between M1 and M2 with high secretion levels of IL-6, TNF-α, PGE2, NO and TGF-β, and gene expression profile related to M1, iNOS, and M2, arginase 1. Furthermore, M0 macrophages cultured with AC or AC-Sp show strong suppression of the macrophage microbicidal function when challenged with S. pneumonia. The presence of PGE2 drives the polarization of M0 macrophages to a M2 profile, and in the presence of COX inhibitors, it reverses the polarization of this macrophage phenotype. The in vivo results demonstrate that the instillation of AC or AC-Sp promotes distinct microenvironments in the lungs of these animals, which influence the resolution of the infection. All these results suggest that the presence of AC or AC-Sp promotes, ...

A melatonina é uma indolamina sintetizada principalmente pela glândula pineal, cuja função está associada à marcação do escuro. Além deste papel cronobiótico, a melatonina também tem papel na defesa e é sintetizada por outros sítios podendo exercer ação parácrina e autócrina, como em células imunocompetentes. Concentrações substancialmente elevadas de melatonina são encontradas no liquido cefalorraquidiano (LCR) que tem sido vinculada à síntese de melatonina por células do sistema nervoso central (SNC), como as microglias. Sabendo-se que estas células são os macrófagos residentes no SNC e que a síntese de melatonina por estes fagócitos já é comprovada, nosso trabalho teve por objetivo avaliar se microglias cerebelares sintetizam essa indolamina e se esta atua potencializando a fagocitose destas células. Nossos resultados mostram que o bloqueio dos receptores de melatonina com o antagonista luzindol, diminuiu tanto a fagocitose induzida por melatonina exógena, quanto à fagocitose basal, indicando que há síntese de melatonina por microglias cerebelares que, por sua vez, age na fagocitose. Esses resultados são relevantes e indicam que a melatonina sintetizada pela microglia, pode estar relacionada com a homeostase do ambiente neural. Sendo assim, nossos dados podem contribuir com estudos que estabeleçam novas estratégias terapêuticas para doenças neuroinflamatórias. / Melatonin is a indolamine synthesized primarily by the pineal gland, whose function is associated with the marking of the dark phase. Beyond this chronobiotic function, melatonin also plays a role in defense and is synthesized by other sites. It may exert paracrine and autocrine action, like in immunocompetent cells. High substantially concentrations of melatonin are found in the cerebrospinal fluid (CSF) that has been linked to the synthesis of melatonin by the central nervous system cells (CNS), such as microglia. Knowing that these cells are the resident macrophages in the CNS and that melatonin synthesis by these phagocytes is proven, our study aims to assess whether cerebellar microglia synthesize this indolamine and whther this acts enhancing phagocytosis of these cells. Our results show that blocking the melatonin receptors with the antagonist, luzindole, both the exogenous melatonin-induced phagocytosis and the basal phagocytosis decreased, indicating that there is melatonin synthesis by cerebellar microglia which acts on phagocytosis. These results are significant and indicate that melatonin synthesized by microglia may be related to the neural environment homeostasis. In this way, our data can contribute, for example, in studies to establish new therapeutic strategies for neuroinflammatory diseases.

Trichophyton rubrum, importante agente de dermatofitoses, é um fungo queratinofílico, capaz de parasitar tecidos como pele e unha. É o principal responsável pelas dermatofitoses crônicas e refratárias ao tratamento e como é uma espécie antropofílica encontra-se muito bem adaptado ao parasitismo humano. Por tratar-se de uma micose cutânea, torna-se necessário o estudo dos fenômenos que ocorrem durante o encontro deste fungo com uma das principais células do sistema imunológico que primeiramente reconhecem o antígeno. Assim sendo, o objetivo deste trabalho é estudar a interação de T.rubrum com macrófagos, para aumentar o conhecimento dos mecanismos envolvidos na resposta imunológica nesta importante patologia. Para isso, foram realizados ensaios de fagocitose de conídios de T.rubrum, seguidos da análise da expressão de moléculas de superfície celular, dosagem de citocinas e viabilidade de macrófagos. Verificamos que o exoantígeno de T.rubrum provocou diminuição da fagocitose de conídios e partículas de zymosan pelos macrófagos. Entretanto, o exoantígeno não interferiu na expressão de moléculas de superfície celular e não foi capaz de estimular os macrófagos a secretar TNF-&#945;, IL-12, IL-10 e óxido nítrico. Já os conídios fagocitados por macrófagos, provocaram diminuição significativa na expressão de suas moléculas de superfície, tais como MHC classe II, CD80 e CD54. Após fagocitose de conídios, os macrófagos foram capazes de secretar uma grande quantidade de TNF-&#945; e IL-10 e após 8 horas de cultivo, os conídios internalizados iniciaram processo de formação de hifa, provocando lise e a conseqüente morte destas células. Por estes achados e pelos estudos prévios já realizados com o T.rubrum, pensamos que a persistência desta infecção fúngica possa estar relacionada com a ação inibitória do fungo sobre os macrófagos, levando à cronicidade observada nestas lesões. / Trichophyton rubrum is the most common pathogen causing dermatophytosis, accounting for approximately 80% of the reported cases of onychomycosis. Since 90% of the chronic dermatophyte infections are caused by T. rubrum, it is likely that this pathogen must have evolved mechanisms that evade or suppress cell-mediated immunity. Several reports have highlighted the participation of phagocytes in the immune defense against fungi; however, few studies have addressed the role of these cells in dermatophytosis. In this study, we investigated the interactions of resident and peritoneal macrophages with T. rubrum. We show here that the interaction of T. rubrum conidia with resident macrophages results in the production of TNF-&#945; and IL-10 but not IL-12 and nitric oxide. Infected macrophages down-regulated the expression of co-stimulatory molecules (CD80 and CD54). We also show that phagocytosis of T. rubrum conidia is inhibited by the addition of fungal exoantigens or mannan. Cytotoxicity assays indicated that after 8 h of conidia ingestion macrophage viability decreased drastically. Electron microscopy revealed that the ingested conidia grow and differentiate into hyphae inside macrophages leading to rupture of the macrophage membrane.