Hepatocyte Permissiveness to Plasmodium Infection Is Conveyed by a Short and Structurally Conserved Region of the CD81 Large Extracellular Domain

Invasion of hepatocytes by Plasmodium sporozoites is a prerequisite for establishment of a malaria infection, and thus represents an attractive target for anti-malarial interventions. Still, the molecular mechanisms underlying sporozoite invasion are largely unknown. We have previously reported that the tetraspanin CD81, a known receptor for the hepatitis C virus (HCV), is required on hepatocytes for infection by sporozoites of several Plasmodium species. Here we have characterized CD81 molecular determinants required for infection of hepatocytic cells by P. yoelii sporozoites. Using CD9/CD81 chimeras, we have identified in CD81 a 21 amino acid stretch located in a domain structurally conserved in the large extracellular loop of tetraspanins, which is sufficient in an otherwise CD9 background to confer susceptibility to P. yoelii infection. By site-directed mutagenesis, we have demonstrated the key role of a solvent-exposed region around residue D137 within this domain. A mAb that requires this region for optimal binding did not block infection, in contrast to other CD81 mAbs. This study has uncovered a new functionally important region of CD81, independent of HCV E2 envelope protein binding domain, and further suggests that CD81 may not interact directly with a parasite ligand during Plasmodium infection, but instead may regulate the function of a yet unknown partner protein

Gene Disruption of Plasmodium falciparum p52 Results in Attenuation of Malaria Liver Stage Development in Cultured Primary Human Hepatocytes

Difficulties with inducing sterile and long lasting protective immunity against malaria with subunit vaccines has renewed interest in vaccinations with attenuated Plasmodium parasites. Immunizations with sporozoites that are attenuated by radiation (RAS) can induce strong protective immunity both in humans and rodent models of malaria. Recently, in rodent parasites it has been shown that through the deletion of a single gene, sporozoites can also become attenuated in liver stage development and, importantly, immunization with these sporozoites results in immune responses identical to RAS. The promise of vaccination using these genetically attenuated sporozoites (GAS) depends on translating the results in rodent malaria models to human malaria. In this study, we perform the first essential step in this transition by disrupting, p52, in P. falciparum an ortholog of the rodent parasite gene, p36p, which we had previously shown can confer long lasting protective immunity in mice. These P. falciparum P52 deficient sporozoites demonstrate gliding motility, cell traversal and an invasion rate into primary human hepatocytes in vitro that is comparable to wild type sporozoites. However, inside the host hepatocyte development is arrested very soon after invasion. This study reveals, for the first time, that disrupting the equivalent gene in both P. falciparum and rodent malaria Plasmodium species generates parasites that become similarly arrested during liver stage development and these results pave the way for further development of GAS for human use

Examination of model predictions at different horizontal grid resolutions

While fluctuations in meteorological and air quality variables occur on a continuum of spatial scales, the horizontal grid spacing of coupled meteorological and photochemical models sets a lower limit on the spatial scales that they can resolve. However, both computational costs and data requirements increase significantly with increasing grid resolution. Therefore, it is important to examine, for any given application, whether the expected benefit of increased grid resolution justifies the extra costs. In this study, we examine temperature and ozone observations and model predictions for three high ozone episodes that occurred over the northeastern United States during the summer of 1995. In the first set of simulations, the meteorological model RAMS4a was run with three two-way nested grids of 108/36/12 km grid spacing covering the United States and the photochemical model UAM-V was run with two grids of 36/12 km grid spacing covering the eastern United States. In the second set of simulations, RAMS4a was run with four two-way nested grids of 108/36/12/4 km grid spacing and UAM-V was run with three grids of 36/12/4 km grid spacing with the finest resolution covering the northeastern United States. Our analysis focuses on the comparison of model predictions for the finest grid domain of the simulations, namely, the region overlapping the 12 km and 4 km domains. A comparison of 12 km versus 4 km fields shows that the increased grid resolution leads to finer texture in the model predictions; however, comparisons of model predictions with observations do not reveal the expected improvement in the predictions. While high-resolution modeling has scientific merit and potential uses, the currently available monitoring networks, in conjunction with the scarceness of highly resolved spatial input data and the limitations of model formulation, do not allow confirmation of the expected superiority of the high-resolution model predictions. © Springer 2005

Examination of model predictions at different horizontal grid resolutions

While fluctuations in meteorological and air quality variables occur on a continuum of spatial scales, the horizontal grid spacing of coupled meteorological and photochemical models sets a lower limit on the spatial scales that they can resolve. However, both computational costs and data requirements increase significantly with increasing grid resolution. Therefore, it is important to examine, for any given application, whether the expected benefit of increased grid resolution justifies the extra costs. In this study, we examine temperature and ozone observations and model predictions for three high ozone episodes that occurred over the northeastern United States during the summer of 1995. In the first set of simulations, the meteorological model RAMS4a was run with three two-way nested grids of 108/36/12 km grid spacing covering the United States and the photochemical model UAM-V was run with two grids of 36/12 km grid spacing covering the eastern United States. In the second set of simulations, RAMS4a was run with four two-way nested grids of 108/36/12/4 km grid spacing and UAM-V was run with three grids of 36/12/4 km grid spacing with the finest resolution covering the northeastern United States. Our analysis focuses on the comparison of model predictions for the finest grid domain of the simulations, namely, the region overlapping the 12 km and 4 km domains. A comparison of 12 km versus 4 km fields shows that the increased grid resolution leads to finer texture in the model predictions; however, comparisons of model predictions with observations do not reveal the expected improvement in the predictions. While high-resolution modeling has scientific merit and potential uses, the currently available monitoring networks, in conjunction with the scarceness of highly resolved spatial input data and the limitations of model formulation, do not allow confirmation of the expected superiority of the high-resolution model predictions. © Springer 2005

Imaging of Plasmodium liver stages to drive next-generation antimalarial drug discovery

Most malaria drug development focuses on parasite stages detected in red blood cells, even though to achieve eradication, next-generation drugs active against both erythrocytic and exo-erythrocytic forms would be preferable. We applied a multifactorial approach to a set of 1 microM), and identified chemical scaffolds with potent activity against both forms. From this screen, we identified an imidazolopiperazine scaffold series that was highly enriched among compounds active against Plasmodium liver stages. Our orally bioavailable lead imidazolopiperazine confers complete causal prophylactic protection (15 mg/kg) in rodent models of malaria and shows potent in vivo blood-stage therapeutic activity. The open source chemical tools resulting from our effort provide starting points for future drug discovery programs, as well as opportunities for researchers to investigate the biology of exo-erythrocytic form

A pre-emptive strike against malaria's stealthy hepatic forms

10.1038/nrd2960Nature Reviews Drug Discovery811854-864NRDD