Sequestration Properties of Blood Stages of <i>P. falciparum</i> in Humans and <i>P. berghei</i> ANKA in Rodents.

<p>Sequestration Properties of Blood Stages of <i>P. falciparum</i> in Humans and <i>P. berghei</i> ANKA in Rodents.</p

Imaging of transgenic <i>P. berghei</i> ANKA parasites in brains of mice ex vivo.

<p>Matched sets of experiments with <i>P. berghei</i> ANKA infections in ECM-sensitive mice (i.e., wild-type mice) or knock-out mice (i.e., IP10^−/−). Knock-out mice do not develop cerebral pathology and this corresponds to a strong reduction in irbc accumulation as compared to infections in wild-type mice (adapted from <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1001032#ppat.1001032-Nie1" target="_blank">[41]</a>). Similar examples of a lack of irbc accumulation can be observed in the brains of mice treated with antibodies against host molecules (e.g., anti-LTβ mAB and anti-CD25 mAB; see <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1001032#ppat.1001032-Amante1" target="_blank">[42]</a>, <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1001032#ppat.1001032-Randall2" target="_blank">[104]</a>). Parasites express GFP::luciferase fusion protein under the control of the eef1a promoter, see <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1001032#ppat-1001032-box003" target="_blank">Box 3</a>). Also, mice infected with a <i>P. berghei</i> ANKA mutant that has had the gene encoding plasmepsin 4 removed do not develop cerebral complications, and again there is a strong reduction of irbc accumulation in the brain of these infected animals (adapted from <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1001032#ppat.1001032-Spaccapelo1" target="_blank">[43]</a>).</p

Imaging of transgenic <i>P. berghei</i> ANKA parasites in vivo and ex vivo.

<p>CD36-mediated sequestration of schizonts in adipose tissue and lungs (adapted from PNAS, 2005 <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1001032#ppat.1001032-FrankeFayard1" target="_blank">[35]</a>). (A, B) Distribution of transgenic <i>P. berghei</i> ANKA parasites, expressing GFP::luciferase fusion protein (<i>ama-1</i> promoter, see <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1001032#ppat-1001032-box003" target="_blank">Box 3</a>). Parasites are visible in lungs, spleen, and adipose tissue in wild-type mice, and principally in the blood circulation and accumulated in the spleen in CD36 knock-out mice. In wild-type mice infected with a non-sequestering K173 line, schizonts are also mainly found in the peripheral blood circulation and accumulated in the spleen (1: adipose tissue; 2: spleen; 3 liver; 4: lungs; 5: heart; 6: kidney; 7: brain). (C) Sequestration of transgenic <i>P. berghei</i> ANKA parasites in microvasculature of adipose tissue (upper panel with under phase contrast and lower panel with GFP-positive schizonts indicated by arrows).</p

<i>P. berghei</i> ANKA asexual blood stage development and expression of proteins in mature schizonts.

<p>(A) In vivo and in vitro development of rings, trophozoites, and schizonts during one cycle of synchronized development. In mice, rings and trophozoites do not sequester but schizonts disappear from the peripheral circulation (upper graph). In vitro schizogony takes place between 18 and 24 hours after invasion of the red blood cell (lower graph). The arrow indicates a multiply infected red blood cell containing three trophozoite-stage parasites; above this cell is a 20-hour schizont (graphs adapted from <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1001032#ppat.1001032-Mons1" target="_blank">[44]</a> and <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1001032#ppat.1001032-FrankeFayard1" target="_blank">[35]</a>. (B) Live mature schizonts of two transgenic lines expressing two different fluorescently tagged PIR proteins either tagged with GFP (eG; PB200064.00.0) or mCherry (mC; PB200026.00.0). These proteins are exported into the cytoplasm of the erythrocyte nucleus stained with Hoechst (H; blue), red blood cell membrane surface protein stained in mC parasites (TER-FITC; green) (J. Braks and B. Franke-Fayard, unpublished data). (C) Live mature schizonts that express GFP and mCherry in the cytoplasm of the merozoites (J. Braks and B. Franke-Fayard, unpublished data).</p

MOESM1 of Transcriptome analysis of Plasmodium berghei during exo-erythrocytic development

Additional file 1: Table S1. RNA-seq data of different life cycle stages of P. berghei used in this study. Table S2. RNA-seq analysis: Raw sequencing counts per gene in the different life cycle stages. Table S3. Gene ontology (GO) term annotation of genes of the individual communities in the GCN. Table S4. P. berghei geneIDs of genes of the different communities in the GCN. Table S5. Genes preferentially expressed in developing EEF stages compared to developing EF stages. Table S6. Genes preferentially expressed in detached cells (DCs/merosomes) compared to erythrocytic schizonts. Table S7. Genes preferentially expressed in blood schizonts (22 h) compared to EEF stages. Table S8. Genes preferentially expressed in blood schizonts (22 h) compared to all other stages. Table S9. Genes preferentially expressed in detached cells (DCs/merosomes) compared to all other stages

MOESM2 of Transcriptome analysis of Plasmodium berghei during exo-erythrocytic development

Additional file 2: Figure S1. Generation and genotyping of parasites expressing gfp under control of the promoter of PBANKA_1003900 (PBANKA_1003900GFP). Figure S2. Fluorescence-activated cell sorting of infected HeLa cells preserved in RNAlater. Figure S3. RNA expression profiles of 3 housekeeping genes (gapdh, actinI, tubulin1). Figure S4. RNA expression profiles of 5 genes encoding serine-repeat antigens, serine-type proteases (SERA1-5). Figure S5. RNA expression profiles of 5 genes encoding proteins of the parasitophorous vacuole membrane (Exported protein 1, Exported protein 2, UIS3, UIS4). Figure S6. RNA expression profiles of genes encoding 2 sporozoite surface proteins (CSP and TRAP). Figure S7. RNA expression profiles of 4 genes encoding enzymes involved in fatty acid biosynthesis (FabB/F, FabI, FabZ, FabG). Figure S8. RNA expression profiles of 6 genes encoding merozoite surface proteins (MSP). Figure S9. RNA expression profiles of 3 genes whose promoter regions have been used to drive expression of fluorescent/luminescent reporter proteins (HSP70, two genes for EF1Îą)

MOESM3 of Transcriptome analysis of Plasmodium berghei during exo-erythrocytic development

Additional file 3. Time laps: GFP expression under the control of PBANKA_1003900 promoter during EEF stage development

Additional file 2 of Streamlining sporozoite isolation from mosquitoes by leveraging the dynamics of migration to the salivary glands

Additional file 2: Figure S2. Increasing yields predicted by (a) mean oocyst densities (yellow line) correspond with increasing uncertainty (yellow shaded area). (b) In groups with higher mean oocyst densities (e.g., 45), estimating the contribution of time may be difficult because of the possibility of pooling individuals with heavily infected midgut (~ 50 oocysts, left pane) consisting of some oocysts that have contributed sporozoites already (arrows) and some still in the process of doing so (arrowheads), with another individual with low infected midgut (4 oocysts, right pane) where the entire contingent of sporozoites have been released (arrowheads). Images were taken at 400 × magnification at 26 days post-blood meal, from individuals whose salivary glands were combined into the same pool

Additional file 4 of Streamlining sporozoite isolation from mosquitoes by leveraging the dynamics of migration to the salivary glands

Additional file 4: Table S2. Sample sizes and measures of parasite infection in the midguts

ICAM-1 is a key receptor mediating cytoadherence and pathology in the Plasmodium chabaudi malaria model

BACKGROUND: Parasite cytoadherence within the microvasculature of tissues and organs of infected individuals is implicated in the pathogenesis of several malaria syndromes. Multiple host receptors may mediate sequestration. The identity of the host receptor(s), or the parasite ligand(s) responsible for sequestration of Plasmodium species other than Plasmodium falciparum is largely unknown. The rodent malaria parasites may be useful to model interactions of parasite species, which lack the var genes with their respective hosts, as other multigene families are shared between the species. The role of the endothelial receptors ICAM-1 and CD36 in cytoadherence and in the development of pathology was investigated in a Plasmodium chabaudi infection in C57BL/6 mice lacking these receptors. The schizont membrane-associated cytoadherence (SMAC) protein of Plasmodium berghei has been shown to exhibit reduced CD36-associated cytoadherence in P. berghei ANKA-infected mice. METHODS: Parasite tissue sequestration and the development of acute stage pathology in P. chabaudi infections of mice lacking CD36 or ICAM-1, their respective wild type controls, and in infections with mutant P. chabaudi parasites lacking the smac gene were compared. Peripheral blood parasitaemia, red blood cell numbers and weight change were monitored throughout the courses of infection. Imaging of bioluminescent parasites in isolated tissues (spleen, lungs, liver, kidney and gut) was used to measure tissue parasite load. RESULTS: This study shows that neither the lack of CD36 nor the deletion of the smac gene from P. chabaudi significantly impacted on acute-stage pathology or parasite sequestration. By contrast, in the absence of ICAM-1, infected animals experience less anaemia and weight loss, reduced parasite accumulation in both spleen and liver and higher peripheral blood parasitaemia during acute stage malaria. The reduction in parasite tissue sequestration in infections of ICAM-1 null mice is maintained after mosquito transmission. CONCLUSIONS: These results indicate that ICAM-1-mediated cytoadherence is important in the P. chabaudi model of malaria and suggest that for rodent malarias, as for P. falciparum, there may be multiple host and parasite molecules involved in sequestration