Eimeria tenella é umas das principais espécies que causam a coccidiose aviária. Para se estudar o perfil de expressão gênico quantitativo em estágios infectantes bibliotecas de LongSAGE foram geradas a partir de merozoítos e esporozoítos. Mais de 35.000 tags foram obtidas, das quais, 9.516 eram únicas. Para a identificação e anotação de genes diferencialmente expressos, as tags foram extraídas, contadas e analisadas estatisticamente por um pacote desenvolvido pelo nosso grupo, SAGE Analysis. Um total de 197 seqüências foram reconstruídas e anotadas automaticamente. Foi observado uma expressão estágio-específica e perfil transcricional distinto entre os estágios. Em merozoítos, foram encontradas proteínas envolvidas na tradução e manutenção da conformação protéica e em esporozoítos, os resultados positivos foram relacionados à cromatina, transporte e atividade catalítica. Para validação da técnica, a expressão diferencial de um pequeno conjunto de genes foi quantificada por RT-qPCR. Os resultados demonstraram uma boa correlação entre estas duas plataformas. / Eimeria tenella is one of the most important causing agents of poultry coccidiosis. To study the quantitative gene expression profile in zoite stages of LongSage libraries were generated from merozoites and sporozoites. More than 35.000 tags were obtained, whose 9.516 were unique. For identification and annotation of differential expressed genes, tags were extracted, counted and submitted to statistical analysis by Sage Analysis, software developed by our group. A total of 197 tags were reconstructed and automatic annotated. Stage-specific expression genes and distinct transcriptional profile were observed between these stages. In merozoites the results were related to protein translation and folding, and in sporozoites the proteins were involved to chromatin structure, transport and catalytic activity. To LongSAGE validation, differential expression was quantified using RT-qPCR to a small group of genes. Good correlation was observed between these platforms.

Eimeria tenella é umas das principais espécies que causam a coccidiose aviária. Para se estudar o perfil de expressão gênico quantitativo em estágios infectantes bibliotecas de LongSAGE foram geradas a partir de merozoítos e esporozoítos. Mais de 35.000 tags foram obtidas, das quais, 9.516 eram únicas. Para a identificação e anotação de genes diferencialmente expressos, as tags foram extraídas, contadas e analisadas estatisticamente por um pacote desenvolvido pelo nosso grupo, SAGE Analysis. Um total de 197 seqüências foram reconstruídas e anotadas automaticamente. Foi observado uma expressão estágio-específica e perfil transcricional distinto entre os estágios. Em merozoítos, foram encontradas proteínas envolvidas na tradução e manutenção da conformação protéica e em esporozoítos, os resultados positivos foram relacionados à cromatina, transporte e atividade catalítica. Para validação da técnica, a expressão diferencial de um pequeno conjunto de genes foi quantificada por RT-qPCR. Os resultados demonstraram uma boa correlação entre estas duas plataformas. / Eimeria tenella is one of the most important causing agents of poultry coccidiosis. To study the quantitative gene expression profile in zoite stages of LongSage libraries were generated from merozoites and sporozoites. More than 35.000 tags were obtained, whose 9.516 were unique. For identification and annotation of differential expressed genes, tags were extracted, counted and submitted to statistical analysis by Sage Analysis, software developed by our group. A total of 197 tags were reconstructed and automatic annotated. Stage-specific expression genes and distinct transcriptional profile were observed between these stages. In merozoites the results were related to protein translation and folding, and in sporozoites the proteins were involved to chromatin structure, transport and catalytic activity. To LongSAGE validation, differential expression was quantified using RT-qPCR to a small group of genes. Good correlation was observed between these platforms.

Immunity to asexual blood-stage of Plasmodium falciparum malaria is believed to be associated with protective antibodies of certain immunoglobulin classes and subclasses. This thesis addressed the importance of antibodies in relation to malaria infection and their effective interactions with Fc gamma receptor IIa (FcyRIIa) polymorphisms. Our data indicate that the frequency of FcyRIIa-R/R131 genotype was statistically significantly higher in Sudanese patients with severe malaria, while the FcyRIIa-H/H131 genotype showed a significant association with mild malaria. The levels of IgG1 and IgG3 subclass antibodies were statistically higher in the mild malaria patients. The Fulani ethnic group in West Africa has been shown to be relatively resistant to malaria. We investigated the possible impact of FcyRIIa polymorphisms in the Fulani and non-Fulani in Mali and Sudan, and analysed their malaria-reactive IgG subclass profiles. The FcyRIIa-H/H131 genotype and H131-allele were found to be prevalent in the Fulani while R131-allele was prevalent in non-Fulani. The Fulani had higher serum levels of IgG1-3, with higher proportion of IgG2 than the non-Fulani. Most clinico-epidemiology studies have been in areas with holo- and hyper-malaria endemicity. We have analysed antibody responses to a panel of six blood-stage antigens in relation to clinical malaria outcome in mesoendemic Sudan. Our results revealed a linear association with anti-AMA-1 IgG1 antibodies and reduced risk of severe malaria while a non-linear relationship with IgG3 antibodies was observed for MSP-2, MSP-3 and GLURP. In the combined final model, the highest levels of IgG1 subclass antibodies to AMA-1, GLURP-R0, and the highest levels of IgG3 subclass antibodies reactive to 3D7 MSP-2 were independently associated with a reduced risk of clinical malaria. Taken together, these data suggest a possible association between FcyRIIa-R/H131 and anti-malarial antibody responses in the clinical outcome of malaria.