<p>Patients with systemic lupus erythematosus (SLE) have an activated type I interferon (IFN) system with an ongoing IFN-α synthesis. This may be caused by circulating immune complexes, consisting of anti-DNA antibodies (Abs) and DNA, with IFN-α inducing capacity. Produced IFN-α may be crucial in the pathogenesis, because this cytokine can break tolerance and promote autoimmunity.</p><p>In the present thesis, possible mechanisms of the IFN-α production in SLE were studied. To investigate whether IFN-α inducing material could be derived from apoptotic cells, IgG from SLE patients (SLE-IgG) were combined with apoptotic cells. This combination induced high IFN-α production in normal peripheral blood mononuclear cells (PBMC). The IFN-α induction was associated to presence of anti-RNP Abs, but not to anti-dsDNA Abs, indicating that two inducers could be active in SLE, one containing DNA and the other RNA.</p><p>Apoptotic cells and SLE-IgG exclusively activated the natural interferon producing cells (NIPC) and the IFN-α response was enhanced by type I IFN and inhibited by IL-10 and TNF-α. The IFN-α induction was dependent on FcγRII, because blocking this receptor reduced IFN-α production and NIPC were found to express FcγRIIa.</p><p>To further elucidate the role of different autoantibodies in the IFN-α induction, sera from patients with Sjögren´s syndrome (SS), containing autoantibodies to RNA binding proteins (SSA, SSB, RNP and/or Sm) were investigated. The combination of SS or SLE sera and apoptotic or necrotic cell material induced high IFN-α production in PBMC. RNA, but not DNA, was required for IFN-α induction, indicating that RNA and Abs to RNA-binding proteins form potent IFN-α inducing complexes.</p><p>The findings in this thesis can explain central mechanisms for the activation of NIPC in SLE, and perhaps also other autoimmune diseases. This activation is mediated by interferogenic immune complexes, and modulating the NIPC activation may be a novel therapeutic approach in SLE.</p>

In sera remitted for anti-nuclear antibody (ANA) analysis, the supplement of a sensitive anti-SSA/Ro ELISA to the conventional ANA screening by immunofluorescence (IF) revealed that one fourth of the individuals with IF-ANA negative, but SSA/Ro ELISA positive sera, had systemic lupus erythematosus (SLE) or cutaneous LE. Consequently, adding a sensitive anti-SSA/Ro ELISA to the ANA screening is valuable for the serological detection of ANA negative SLE/LE patients. SLE patients often have measurable interferon-alpha (IFN-α) levels in serum, and IFN-α treatment of patients with non-autoimmune diseases can induce SLE. Thus, the type I IFN system seems to be important in SLE and was therefore investigated. Initially, a decreased IFN-α producing capacity, due to a 70-fold reduction in the number of circulating natural IFN-α producing cells (NIPC), was noted in peripheral blood mononuclear cells (PBMC) from SLE patients. SLE-sera contained an endogenous IFN-α inducing factor (SLE-IIF), consisting of IgG and DNA in the form of small immune complexes (300-1000 kD). The SLE-IIF selectively activated NIPC and was more common in sera from patients with active disease compared to individuals with inactive disease. IFN-α producing cells could be detected by immunohistochemistry in both lesional and unaffected skin from SLE patients, and IFN-α gene transcription could be verified by in situ hybridisation in some of the skin biopsies. A reduced number of NIPC, detected by expression of the blood dendritic cell antigen (BDCA)-2, was noted among SLE-PBMC. The IFN-α production triggered by SLE-IIF in SLE-PBMC was inhibited by monoclonal antibodies (mAbs) to BDCA-2 and markedly decreased by anti-BDCA-4 mAbs. The observations in the present thesis may explain the ongoing IFN-α production in SLE patients, indicate an important role for the activated type I IFN system in the pathogenesis, and suggest that direct targeting of SLE-NIPC may constitute a new therapeutic principle in SLE.

Patients with systemic lupus erythematosus (SLE) have an activated type I interferon (IFN) system with an ongoing IFN-α synthesis. This may be caused by circulating immune complexes, consisting of anti-DNA antibodies (Abs) and DNA, with IFN-α inducing capacity. Produced IFN-α may be crucial in the pathogenesis, because this cytokine can break tolerance and promote autoimmunity. In the present thesis, possible mechanisms of the IFN-α production in SLE were studied. To investigate whether IFN-α inducing material could be derived from apoptotic cells, IgG from SLE patients (SLE-IgG) were combined with apoptotic cells. This combination induced high IFN-α production in normal peripheral blood mononuclear cells (PBMC). The IFN-α induction was associated to presence of anti-RNP Abs, but not to anti-dsDNA Abs, indicating that two inducers could be active in SLE, one containing DNA and the other RNA. Apoptotic cells and SLE-IgG exclusively activated the natural interferon producing cells (NIPC) and the IFN-α response was enhanced by type I IFN and inhibited by IL-10 and TNF-α. The IFN-α induction was dependent on FcγRII, because blocking this receptor reduced IFN-α production and NIPC were found to express FcγRIIa. To further elucidate the role of different autoantibodies in the IFN-α induction, sera from patients with Sjögren´s syndrome (SS), containing autoantibodies to RNA binding proteins (SSA, SSB, RNP and/or Sm) were investigated. The combination of SS or SLE sera and apoptotic or necrotic cell material induced high IFN-α production in PBMC. RNA, but not DNA, was required for IFN-α induction, indicating that RNA and Abs to RNA-binding proteins form potent IFN-α inducing complexes. The findings in this thesis can explain central mechanisms for the activation of NIPC in SLE, and perhaps also other autoimmune diseases. This activation is mediated by interferogenic immune complexes, and modulating the NIPC activation may be a novel therapeutic approach in SLE.

Le virus de l’hépatite C (VHC) est un problème mondial. La majorité des personnes infectées (70-85%) développent une infection chronique qui cause des complications hépatiques. Le seul régime thérapeutique approuvé pour le VHC est l'interféron alpha (IFN-α). Ce traitement a un taux de réussite de 50-80% selon le génotype de virus et le moment de l'initiation de la thérapie. Les facteurs régissant la réponse au traitement ne sont pas bien définis. Des études antérieures ont suggéré un rôle potentiel de la réponse immunitaire de l'hôte au succès de la thérapie, toutefois, ces résultats sont controversés. Nous avons émis l'hypothèse que la réponse immunitaire de l’hôte sera plus efficace chez les patients qui commencent la thérapie tôt pendant la phase aiguë de l'infection. En revanche, la réponse immunitaire sera épuisée lorsque le traitement est initié pendant la phase chronique. L'objectif principal de ce mémoire est d’étudier les facteurs immunologiques qui régissent la réponse à la thérapie, et de déterminer si la contribution de la réponse immunitaire de l'hôte peut être influencée par la période de l'infection. Nos résultats démontrent l'efficacité de la restauration de la réponse immunitaire spécifique au VHC lorsque la thérapie par l'interféron est initiée tôt. Ceci est démontré par le sauvetage des cellules T efficaces spécifiques au VHC efficace similaires à celles observées chez les individus qui ont résolu spontanément, suggérant ainsi qu'elles jouent un rôle actif dans la réponse au traitement. Toutefois, cette réponse n'a pas été restaurée chez les patients traités au cours de la phase chronique. Ces résultats ont des implications importantes dans la compréhension des mécanismes sous-jacents à la réponse aux traitements actuels et au développement des nouvelles thérapies. / Hepatitis C virus (HCV) is a major public health problem worldwide. Only 15-30% of infected individuals clear the virus spontaneously, while the majority develops chronic infection that causes liver complications. The only approved therapy for HCV is interferon alpha (IFN-α) based. This therapy has a 50-80% success rate depending on the infecting virus genotype and the timing of initiation of therapy. Factors governing the response to therapy are not well defined. Previous studies have suggested a role for the host immune response in the success of therapy. However, these results were controversial. We hypothesized that host immunity has an effective role in the success of IFN-α therapy when initiated early during the acute phase of HCV infection, while late initiation during the chronic phase minimizes this role. The main objective of this thesis was to dissect the immunological factors governing the differential response to IFN-α therapy, and to determine if the contribution of the immune response to success of therapy might be influenced by the period of infection. Our results demonstrate restoration of efficient HCV-specific immune responses when therapy is initiated early during the acute phase. This is demonstrated by the rescue of functional HCV-specific T cells similar to those observed in spontaneously resolved individuals, suggesting that they may play an active role in response to therapy. However, such responses were not restored following late therapy suggesting irreversible damage to the host’s defence system with chronicity. These findings have important implications in understanding the mechanisms underlying response to current treatments and development of novel therapies.

Diss. (sammanfattning) Stockholm : Karolinska institutet, 2005. / Härtill 5 uppsatser.

Made available in DSpace on 2015-07-22T20:49:40Z (GMT). No. of bitstreams: 0 Previous issue date: 2011-07-27 / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / Introdução: Pacientes com Hepatite C frequentemente tendem a produzir autoanticorpos. No teste fator antinúcleo em células HEp-2 (ANA-HEp-20), esses autoanticorpos geram diversos padrões de imunofluorescência, sendo o nuclear pontilhado fino o mais frequente deles. Recentemente, tem sido descrito um novo padrão de ANA-HEp-2 em pacientes com HCV, denominado padrão Rods e Rings (R&R), caracterizado por anéis e bastões. Objetivos: Avaliar as características clínicas, virológicas e padrão de resposta terapêutica dos pacientes que apresentam autoanticorpos que geram o padrão R&R, bem como proceder a uma avaliação preliminar dos aspectos celulares e moleculares desse novo sistema de autoantígenos. Métodos: Amostras de soro coletadas de 1998 até 2008 de 597 pacientes foram submetidas ao teste de ANA-HEp-2 em lâminas Euroimmun ou INOVA e classificados como R&R positivos quando apresentaram fluorescência sob forma de bastões de 3 a 10μm de comprimento e anéis de 2 a 5μm de diâmetro no citoplasma das células HEp-2. Entre os pacientes testados, 342 tinham HCV e 200 tinham outras doenças hepáticas crônicas ou autoimunes reumáticas, além de 55 pacientes co-infectados com HCV+HIV. As informações clínicas, virológicas e terapêuticas foram coletadas de bancos de dados atrelados às amostras de soro dos pacientes. Células HEp-2, 3T3 e MH22A foram cultivadas normalmente e/ou submetidas à tratamentos in vitro (tripsina e ribavirina) e com dois métodos alternativos de fixação, para estudo das estruturas do R&R por imunofluorescência indireta simples ou com técnicas de dupla-marcação com amostras R&R-positivas e anticorpos antitubulina-alfa e anti-CTP-sintase. Resultados: Dos 342 pacientes com HCV, 51 (15%) apresentaram o padrão R&R, enquanto que dos 200 pacientes com outras doenças hepáticas ou autoimunes, apenas um apresentou o padrão R&R (p<0.0001). Dos pacientes com HCV, 174 eram tratados e 168 não tratados. Dos 174 tratados, 49 (28%) apresentaram o padrão R&R contra apenas dois (1%) dos não tratados (p<0.0001). De 134 tratados e com informação adequada sobre a medicação utilizada, 108 tomavam interferon-α e ribavirina e 23 tomavam apenas interferon-α. Quarenta e um (38%) dos 103 que tomavam interferon-α e ribavirina apresentaram o padrão R&R contra nenhum (0%) dos 23 que tomavam apenas interferon-α (p= 0.0001). Quanto aos outros padrões de ANA-HEp-2, dos 23 pacientes que tomavam apenas interferon-α, 12 (52%) foram positivos enquanto apenas 27 (25%) dos que tomavam interferon-α e ribavirina foram positivos (p= 0.010). 9% dos pacientes co-infectados com HCV+HIV apresentaram o padrão R&R. Não encontramos relação entre a presença do padrão R&R e o genótipo do vírus, a carga viral e os dados demográficos dos pacientes com HCV. A porcentagem de respondedores ao tratamento foi ligeiramente menor nos pacientes que apresentaram o R&R, porem sem atingir nível de significância estatística (p=0,150). Lâminas ANA-HEp-2 de algumas marcas comerciais que não Euroimmun e INOVA e aquelas elaboradas no próprio laboratório não apresentaram as estruturas do R&R. Quando tratadas in vitro com ribavirina ou tripsina, as células HEp-2 ou 3T3 e MH22A de camundongo cultivadas expressaram vários anéis e bastões reconhecidos pelas amostras de soro R&R-positivas. Não observamos colocalização das estruturas R&R com microtúbulos e observamos fraca colocalização dos anéis e bastões com CTP-sintase. Conclusões: Autoanticorpos associados ao padrão R&R ocorreram em íntima associação ao uso de interferon-α e ribavirina em pacientes com hepatite HCV, independentemente de serem portadores do HIV. Não houve associação às características demográficas dos pacientes, ao perfil de resposta terapêutica, ao genótipo do HCV ou à carga viral. As estruturas em anéis e bastões associadas ao padrão R&R não ocorrem nas condições normais avaliadas, podendo ser induzidas in vitro pela exposição à ribavirina ou à tripsina. Há algum grau de conservação filogenética dos autoantígenos associados ao padrão R&R. Evidências preliminares indicam a presença da enzima CTP-sintase nas estruturas em anéis e bastões reconhecidas pelos autoanticorpos humanos. / Introduction: Patients with Hepatitis C frequently produce autoantibodies. In the antinuclear antibody assay on HEp-2 cells (ANA-HEp-2) these autoantibodies generate several immunofluorescence patterns, and the nuclear fine speckled pattern is the most common. A novel ANA-HEp-2 pattern has been recently reported, characterized by the presence of rods and rings in the cytoplasm. Objectives: To study the clinical and virological features, as well as the profile of therapeutic response of patients presenting autoantibodies generating the rods and rings (R&R) ANA-HEp-2 pattern and to perform a preliminary analysis of the cellular and molecular aspects of this novel autoantigen system. Methods: Serum samples obtained from 1988 to 2008 from 597 patients were processed in the ANA-HEp-2 assay on Euroimmun or INOVA slides and classified as R&R-positive when presenting immunofluorescence as 3-10μm long rods and 2-5μm diameter rings in the cytoplasm of HEp-2 cells. Among the tested patients, 342 had HCV, 200 had other chronic liver diseases or rheumatic autoimmune diseases, and 55 had HCV and HIV. Clinical, virological, and treatment information was obtained from the clinical data bank. Human HEp-2, and murine 3T3, and MH22A cell lines were cultured as usual and under special stimuli (exposure to trypsin or ribavirin), and prepared with alternative fixation protocols for processing in single or double indirect immunofluorescence with human anti-R&R serum and antibodies to tubulin and to CTP-synthase. Results: Among the 342 HCV patients, 51 (15%) presented the R&R pattern as opposed to only one among the 200 patients with other liver diseases and autoimmune rheumatic diseases (p<0.001). Among the HCV patients, 174 had been treated and 168 had received no treatment. Among the 174 treated patients, 49 (28%) presented the R&R pattern as opposed to only two (1%) of the 168 non-treated HCV patients (p<0.001). Among 134 HCV treated patients with detailed information on the treatment protocol, 108 used interferon-α and ribavirin and 23 used only interferon-α. Forty-one (38%) of the R&R pattern as opposed to none of the 23 patients receiving only interferon-α (p=0.0001). In contrast, 12 (52%) of the 23 patients receiving only interferon-α presented other non-R&R ANA-HEp-2 patterns as opposed to only 27 (25%) of the 108 patients under interferon-α and ribavirin (p=0.01). 9% of the patients with HCV and HIV presented the R&R pattern. There was no association between the occurrence of the R&R pattern and HCV genotype, viral load, and demographic features. The frequency of sustained virologic response was slightly lower in the R&R-positive patients but the difference did not reach statistical significance (p=0.15). ANA-HEp-2 slides from brands other than Euroimmun and INOVA, as well as in-house produced slides did not express the R&R structures. When treated in vitro with ribavirin or trypsin, HEp-2 cells and murine 3T3, and MH22A cell lines expressed prominent R&R structures recognized by human HCV serum samples. There was partial weak colocalization of CTP-synthase in the R&R structures but no colocalizadion of microtubule. Conclusions: autoantibodies associated with the R&R pattern were strongly associated with the use of interferon-α and ribavirin in patients with HCV, independently of co-infection with HIV. There was no association with demographic characteristics, the profile of therapeutic response, HCV genotype, and viral load. The rods and rings structures associated with the R&R pattern did not occur under normal conditions, but could be induced in vitro by exposure to ribavirin or trypsin. There is some degree of phylogenetic conservation of the autoantigens associated to the R&R pattern. Preliminary evidence indicates the presence of CTP-synthase in the cytoplasmic rods and rings structures recognized by human autoantibodies from HCV patients. / FAPESP: 2009/03796-5 / FAPESP: 2010/50710-6 / TEDE / BV UNIFESP: Teses e dissertações

Submitted by Priscila Nascimento (pnascimento@icict.fiocruz.br) on 2012-11-21T16:17:01Z No. of bitstreams: 1 carla-franca-wolanski-de-almeida.pdf: 1099876 bytes, checksum: e454fb4f1e8e4f2a5e837e602abcf47e (MD5) / Made available in DSpace on 2012-11-21T16:17:01Z (GMT). No. of bitstreams: 1 carla-franca-wolanski-de-almeida.pdf: 1099876 bytes, checksum: e454fb4f1e8e4f2a5e837e602abcf47e (MD5) Previous issue date: 2009 / Fundação Oswaldo Cruz. Instituto de Tecnologia em Imunobiológicos. Rio de Janeiro, RJ, Brasil / Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Rio de Janeiro, RJ, Brasil. / Controle em processo é parte das ações para garantia da qualidade dos produtos a fim de que eles atendam aos requisitos mínimos estabelecidos para seu uso. Os resultados obtidos nos ensaios orientam na tomada de decisões, bem como em ajustes ao processo de produção. Muitas vezes tais resultados são condições imperativas para as etapas subsequentes. O monitoramento do processo produtivo se dá a partir do estabelecimento dos pontos críticos de controle, que precisam ser verificados a cada lote, pois interferem diretamente na qualidade e rendimentos da produção. Portanto, a confiabilidade e agilidade deresposta são fundamentais para correta tomada de decisão e/ou ajuste. O objetivo deste trabalho é propor um modelo de laboratório de controle em processo nos aspectos gerenciais e principalmente nos aspectos técnicos aplicados ao processo de produção do biofármaco Interferon alfa 2b humano recombinante. Este produto, oferecido no portfólio de Bio-Manguinhos encontra-se em processode transferência de tecnologia com o Instituto cubano CIGB. Com base nos dados da literatura foram estabelecidas as principais etapas do processo produtivo a para expressão de proteínas heterólogas, em sistema recombinante utilizando-se a bactéria Escherichia coli. Os pontos críticos de controle foram estabelecidos utilizando-se a metodologia de análise de riscos HACCP (Hazard Analisys Critical Control Point). Para as análises de controle foram propostas metodologias analíticas que proporcionem a consistência de resultados e que possam garantir a qualidade final do processo de manufatura do produto. O modelo proposto também enfatiza a conduta de ações rotineiras de forma que qualquer desvio possa ser imediatamente identificado e suas causas apuradas além de executar as análises com segurança. Os critérios gerenciais e técnicos abordados foram avaliados criticamente, utilizando-se os procedimentos do sistema de garantia da qualidade vigentes no Instituto, bem como na estrutura do laboratório de controle em processodestinado à vacina contra Hib, que foi adotado como premissa básica, fruto da transferência de tecnologia bem sucedida em Bio-Manguinhos. A proposta elaborada poderá ser utilizada em outros laboratórios destinados ao controle em processo de novos produtos em Bio-Manguinhos e enfatiza os critérios técnicos como condicionais para confiabilidade de medição. / In-process control is a part of the actions to guarantee the quality for final products in order to attend the minimum requirements established for their using. The results obtained from analytical assays guide on decision-making as well as production process adjustments. Often, these results are conditioned to the follow the subsequent stages. The process monitoring occurs from the establishment of critical control points to verify in each batch because theydeterminate product quality and process yield. Therefore, the reliability and speedof response are critical for correct decision-making and adjustment. The objective of this work is to propose a model in process control laboratory in management aspects and especially the technical aspects applied to the production of human recombinant interferon alpha 2b biopharmaceutical, object of this study. Bio-Manguinhos supplies this product in its portfolio, which production technology is being transferred from the CIGB, a Cuban Institution. Based on literature data the main steps were established for a production process of heterologous proteins expressed using a recombinant system with the Escherichia colibacteria. The critical control points were established using the risk assessment methodology HACCP, Hazard analysis Critical Control, HACCP. Analytical methodologies were proposed to improve results consistency and assure the final quality of production process. The proposed model emphasizes the routine actions, so that, any deviation could be immediately identified, their causes verified and activities performed reliably. The management and technical criteria described have been critically evaluated using the Bio-Manguinhos institute quality assurance procedures as well as compared to the structure of the in-process control laboratory for Hib vaccine. This laboratory, adopted as basic premise, resulted from a successful technology transfer in Bio-Manguinhos. The proposal drawn up can be used in other laboratories for in- process control in new products in Bio-Manguinhos and emphasizes the technical criteria for conditional measurement reliability.

Submitted by Priscila Nascimento (pnascimento@icict.fiocruz.br) on 2012-11-26T11:51:30Z No. of bitstreams: 1 luciana-de-santos-almeida.pdf: 1492447 bytes, checksum: fb4ac7dc61ce8aa49ecd3124c10c4ec4 (MD5) / Made available in DSpace on 2012-11-26T11:51:30Z (GMT). No. of bitstreams: 1 luciana-de-santos-almeida.pdf: 1492447 bytes, checksum: fb4ac7dc61ce8aa49ecd3124c10c4ec4 (MD5) Previous issue date: 2009 / Fundação Oswaldo Cruz. Instituto de Tecnologia em Imunobiológicos. Rio de Janeiro, RJ, Brasil / Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Rio de Janeiro, RJ, Brasil. / A Era da Engenharia Genética ou Biotecnologia Moderna, que teve início nos anos 1970, encontra diversas aplicações em vários segmentos de atividades, dentre eles a saúde e alimentos. Na área de saúde têm -se o destaque para a indústria de biofármacos, que tem tido um crescimento acelerado nos últimos anos, pelo fato desses novos medicamentos serem voltados para a terapêutica de doenças que aflige uma parcela considerável da população mundial. Como exemplo pode-se citar o interferon alfa 2b humano recombinante, que faz parte do Programa de Medicamentos de Dispensação Excepcional do Ministério da Saúde, e está incluído no Protocolo Clínico para Hepatite C. Como forma de atender à demanda da população brasileira quanto às necessidades desse biofármaco, Bio-Manguinhos assinou em 2004 um Contrato de Transferência de Tecnologia como o Centro de Engenharia Genética e Biotecnologia de Cuba (CIGB), com o propósito de nacionalizar a produção do referido medicamento. Este trabalho teve como objetivo realizar uma pesquisa qualitativa para elencar os Requerimentos Fundamentais para Transferência, Registro Sanitário, Estabelecimento e Parâmetros de Estabilidade de Bancos de Células de E. coli que expressa o interferon alfa 2b humano recombinante. Foi elaborado um questionário, que serviu como fonte de coleta de dados, que foi enviado por e-mail a diversas Agências Regulatórias e pesquisadores da academia científica. Esse questionário teve como objetivo elencar os principais testes/requisitos que os Bancos de Células devem apresentar. O índice geral de retorno dos questionários foi igual a 39,3%, sendo que o índice de respostas dos pesquisadores foi igual a 62,5% e das Agências Regulatórias igual a 8,3%. O índice de marcação dos itens ficou em torno de 80% e ficou estabelecido que aqueles itens que tiveram um índice maior que 50% seriam os requerimentos indispensáveis a serem descritos. Não foi possível validar o trabalho pelo índice de retorno dos questionários por parte das Agências Regulatórias. Porém, acredita-se que aquilo que foi descrito pode servir como um passo inicial na elaboração de uma norma específica relativa ao Controle de Banco de Células de procariotos, pois representa a opinião de experientes pesquisadores da comunidade científica. E ainda contribuir nos requisitos específicos que serão futuramente estabelecidos pela ANVISA, relativos ao registro de um produto biogenérico/biossimilar. / The Age of Genetic Engineering and Modern Biotechnology, which began in the 1970s, has had several applications in various business activities, among of which are health and food. In health area the emphasis has been on the biopharmaceutical industry that has had a rapid growth in recent years, because these new drugs are aimed towards the therapy of diseases that afflict a significant part of the world’s population. As an example, it can be mentioned the recombinant human interferon-alpha 2b, which is part of the Drugs of Exceptional Dispensation Program of the Ministry of Health, also is it in the Clinical Protocol for Hepatitis C. In order to meet the demand of the Brazilian population for this drug, in 2004, Bio-Manguinhos signed an Agreement on Technology Transfer with the Center for Genetic and Biotechnology Engineering of Cuba (CIGB), so that this medicine could be produced at national level. This study aimed at doing a qualitative research to list the Fundamental Requirements for Transfer, Registration, Establishment and Stability Parameters of Banks of E. coli Cells expressing recombinant human interferon-alpha 2b. A questionnaire was compiled for data collection. It was sent by e-mail to a number of regulatory agencies and researchers of the scientific academy. This questionnaire aimed at listing the main tests / requirements that banks must submit Cells. The overall rate of return of questionnaires was 39.3%, and the rate of responses from researchers was 62.5% and from regulatory agencies was 8.3 %. The rate of items marked was around 80%, then it was agreed that those items that had a rate higher than 50% would be the essential requirements to be described. The work was not validated due to the low rate of return of questionnaires from the Regulatory Agencies. However, it is believed that what was described can serve as the first step in developing a specific standard relating to control the Banks of Prokaryotes Cells, since it represents the expertise opinion of researchers in the scientific community. Yet, it contributes to the specific requirements that shall eventually be established by ANVISA for the registration of a biogeneric / biosimilar product.

No description available.

INTRODUÇÃO: A mielopatia associada ao HTLV-1/paraparesia espástica tropical (HAM/TSP) é a manifestação neurológica mais descrita da infecção pelo HTLV-1. Embora o risco de seu desenvolvimento ao longo da vida do portador dessa retrovirose seja pequeno, 2% a 5%, tal afecção pode causar um importante comprometimento funcional e prejuízo na qualidade de vida. Dadas as exigências atuais por melhores cuidados de saúde, os profissionais de saúde frequentemente empregam diferentes terapêuticas disponíveis para tratar pacientes com HAM/TSP, dentre os quais se destacam as drogas imunomoduladoras, visando reduzir a incapacidade e melhorar a qualidade de vida dos pacientes acometidos. Porém, ainda não há consenso sobre qual seria a melhor intervenção e, quando indicado, qual o melhor imunomodulador para o tratamento da HAM/TSP. MÉTODOS: Com o propósito de buscar-se evidência da eficácia dos medicamentos imunomoduladores no tratamento da HAM/TSP, foi conduzida uma revisão sistemática de ensaios clínicos randomizados e controlados nas bases de dados MEDLINE, LILACS, Embase e Registro Cochrane de Ensaios Clínicos Controlados. Selecionaram-se estudos relevantes, sem restrição de idioma e publicados até em 31/01/13. Além disso, foi realizada busca manual das referências bibliográficas dos estudos selecionados. RESULTADOS: Três ensaios clínicos randomizados e controlados que preencheram os critérios de inclusão foram analisados: dois deles abordando a eficácia do interferon alfa e um de zidovudina associada à lamivudina. CONCLUSÕES: Os pacientes que receberam 3,0 milhões de UI de interferon alfa, pela via intramuscular, uma vez por dia, durante 28 dias consecutivos, apresentaram melhora temporária do comprometimento funcional e do distúrbio urinário, quando comparados aos que receberam 0,3 ou 1,0 milhão de UI de interferon alfa, e apresentaram efeitos adversos toleráveis e temporários. A intensidade da melhora e o tempo do benefício dependeram do grau de comprometimento funcional e urinário prévio ao estudo. Os pacientes que receberam 300 mg de zidovudina e 150 mg de lamivudina, pela via oral, duas vezes por dia, durante 24 semanas, não apresentaram melhora significativa do comprometimento funcional, da dor em membros inferiores ou do distúrbio urinário, quando comparados aos pacientes que receberam placebo. A literatura revisada mostrou-se escassa e heterogênea no que se refere às drogas utilizadas e à dosagem, bem como nos desfechos clínicos avaliados. Em função disso, não se realizou metanálise. Conclui-se que não há evidência definitiva de eficácia do uso de imunomoduladores para o tratamento da HAM/TSP / BACKGROUND: HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP) is the most frequently described neurological manifestation of this retroviral infection. Even through the lifetime risk of HTLV-1 carries developing HAM/TSP is low (2% to 5%) this condition may cause significant disability and compromise quality of life. Given current demands for better health care, health professionals often use different available therapies to treat patients with HAM/TSP, and, particularly, immunomodulating drugs to reduce disability and improve quality of life. However, so far there is no consensus about the best intervention, and, when recommended, the best immunomodulating drug for the treatment of HAM/TSP. METHODS: To establish whether there is evidence of the efficacy of immunomodulating drugs in the treatment of HAM/TSP. We conducted a systematic review of randomized, double-blind, controlled trials, searching MEDLINE, LILACS, EMBASE and the Cochrane Central Register of Controlled Trials. We selected relevant studies published until January 31st, 2013, regardless of language. In addition, a manual search of the references of selected studies was conducted. RESULTS: Three randomized controlled clinical trials that met our inclusion criteria were analyzed: two of them address the efficacy of interferon alfa and another of zidovudine plus lamivudine. CONCLUSIONS: Patients who received 3.0 million IU of interferon alfa, intramuscularly, once a day, for 28 consecutive days, showed temporary improvement of functional impairment and urinary disorder, as compared to those who received 0.3 or 1.0 million IU of interferon alfa, and exhibited with tolerable and temporary side effects. The intensity and duration of improvement depended on the degree of functional impairment and urinary disorder prior to the study. Patients who received 300 mg zidovudine plus 150 mg lamivudine, orally, twice a day, for 24 weeks showed no significant improvement in functional impairment, in lower limbs pain or in urinary disorder when compared to patients who received placebo. The literature we reviewed is scarce and heterogeneous in terms of used drugs and in evaluated clinical outcomes. Therefore, a meta-analysis was not performed. We concluded that there is no definitive evidence of the efficacy of imunnomodulating drugs in the treatment of HAM/TSP