Project description:During the progress through its complex life cycle, the malaria parasite requires strict control of gene expression but the mechanism controlling this process remain poorly understood and unexplored for therapeutic purposes. A group of the apicomplexa-specific putative transcription factors with AP2 DNA binding domain (apiAP2) has been identified as the major regulators of the parasite transcriptome at multiple stages. The targets and the role of the majority of the members of the family however are unknown.
Project description:The transmission of the malaria parasite between mosquitoes and mammals requires translational repression to ensure that only the proper proteins are expressed and correct folded at the right time, it will need for the next developmental stage.Due to their essential role for malaria transmission, gametocytes represent prime targets for transmission-blocking strategies intended to prevent spread of the deadly disease. In this study, we generate HspJ62 gene knockout line (ΔHspJ62) that is gametocyte non-producing lines. Transcriptional profiling of wild type Plasmodium berghei and genetic KO Hspj62 genes at single time points during erythrocytic parasite development.
Project description:During the malaria infection, Plasmodium parasites invade the host’s red blood cells where they can differentiate into two different life forms. The majority will replicate asexually and infect new erythrocytes. A small percentage, however, will transform into gametocytes – a specialized sexual stage able to survive and develop when taken up by Anopheles mosquito. As the gametocytes ensure the parasite’s transmission to a new host, their generation is an attractive target for new antimalarial interventions. The molecular mechanisms controlling gametocytogenesis, however, remain largely unknown due to the technical challenges: the early gametocytes are morphologically indistinguishable from asexual parasites and present in very low numbers during the infection. Recently, AP2-G - a transcription factor from an apicomplexa-specific apiAP2 family – was described as indispensable for gametocyte commitment in both human malaria parasite Plasmodium falciparum and rodent malaria model Plasmodium berghei. Therefore, we have decided to test whether the overexpression of this factor alone could increase gametocyte production and enable the investigation of uncharacterised, earliest stages of gametocyte development. To this end, we have engineered PBGAMi - a Plasmodium berghei line, in which all parasites were ap2-g deficient by default but able to overexpress it when induced with rapamycin. While the control parasites (PBGAMi R-), as expected, differentiated into asexual forms (schizonts) only, almost all rapamycin-treated parasites (PBGAMi R+) transformed into gametocytes. We used the generated line to perform RNA-seq analysis of the R- and R+ populations at different time points of their development and identify the changes arising between them, mapping the sequence of events leading to the formation of gametocytes.
Project description:In malaria, T cells play a dual role by both restricting parasite growth and mediating immunopathology such as the deadly neuroinflammation called cerebral malaria. During experimental cerebral malaria (ECM), IFN produced by CD4 T cells promotes CD8 T cell sequestration in brain capillaries, resulting in endothelial damage, oedema and death. However the antigen-presenting cells controlling the development of CD4 T cell responses, as well as the antigens recognized by these CD4 T cells, are unknown. Here we used mass spectrometry to characterize the MHC II immunopeptidome presented by dendritic cells during blood stage malaria in C57BL/6 mice. We identified 14 MHC II ligands derived from 13 conserved Plasmodium berghei proteins that we validated in vivo. This work profiles the first MHC II immunopeptidome in a mouse model of blood stage malaria.