Evelyn Gitau

Modern molecular typing techniques now permit to identify rapidly var genes encoding dominant expressed PfEMP-1 variants in a given isolate and allow for the first time to define cellular immune responses to that variant. As a post doc in Britta Urban's group, I use the recombinant DBLα tags expressed from acute malaria patient samples to determine the proportion of CD4+ T cells that secrete IFNg, IL10, IL4 and IL2 by 9-colour FACS during an acute infection and after convalescence. Ultimately, we want to define if and how the phenotypic and antigenic properties of a given parasite isolate shape the T-cell and B-cell responses that underlie the production of short-lived and long-lived antibodies against PfEMP-1.

Recently we have begun to investigate the potential role of biomarkers both as indicators of disease classification and also as a means of better understanding the patho-physiology. It is accepted that the pathogenesis of severe malaria will be associated with "compounds" originating from host or pathogen as part of the disease process that can be used to characterise the disease state of the individual. We have used proteomics to identify potential biomarker proteins in cerebral spinal fluid (CSF) and plasma from children with CM, acute bacterial meningitis (ABM) and other encephalopathies. The working hypothesis is that studying the plasma and CSF proteome may provide insights into the biochemical perturbations induced by the parasite including affects on specific neuronal populations, and clarify the basic molecular basis of the pathophysiological changes observed in patients. The initial studies utilizing various proteomic strategies including two dimensional gel electrophoresis (2D SDS PAGE) and various liquid chromatography platforms coupled to tandem mass spectrometry, yielded potential protein biomarkers that could be used in the diagnosis and treatment of disease. In total over 500 hundred proteins both from the host and pathogen were definitively identified with unique patterns of proteins being found with potential diagnostic value for each of the three conditions. We are now carrying out downstream data analysis to validate the potential biomarkers identified as well as create a peptide library of potential antigens.