These reports suggest that some protozoans acquire host sterols, raising the prospect that sterol supply to them may be a potential therapeutic target in these pathogens. An association between cholesterol and malaria parasites, as reported in a patient-based study, showed that high density lipoprotein HDL levels in serum decreased markedly in patients with Plasmodium vivax infections Lambrecht et al.
While malaria parasites could obtain sterols in various ways, blood-stage Plasmodium species possibly take up lipoproteins as a sterol source. However, the known major lipoprotein receptors, including class B scavenger receptors SR-Bs , are generally scarce on mature erythrocyte surfaces Edelman et al. Additionally, no candidate genes have been identified in the P.
These findings indicate that foreign lipoprotein receptors are required for lipoprotein delivery to pRBCs during parasite growth. In this study, we explored possibilities for the lipoprotein receptors on the pRBCs and found that host exosomes delivered CD36 selectively to the pRBC membranes.
Health was monitored every 12 h, and blood sampling was performed from tail veins every 48 h. Parasitemia was estimated by Giemsa-stained thin blood smears.
Parasitemia was estimated from thin blood smears stained with Giemsa. The experiments using human erythrocytes were performed under the guidelines of the ethics committee of The University of Tokyo and The reaction was quenched using 0. Erythrocytes were washed in 0. The images were captured by Zeiss LSM, a confocal microscopy system.
After washing in DMEM containing 0. Erythrocytes were washed with RPMI media containing 0. To minimize platelet contamination, mouse whole blood collected with EDTA was first centrifuged under a 1. After suspension in PBS 2. CMK cells, suspended at 1. The culture medium was harvested, passed through a 0.
The DiI-labelled exosomes were isolated by sequential ultracentrifugation steps as described above. Erythrocytes were washed three times with RPMI media containing 0. Smears were prepared from them for further analyses. DiI-labelled exosomes 0. The 7-week-old recipient male Kusabira-Orange mice were X-ray irradiated total 9. At 5 weeks post-transplantation, KuO fluorescence was confirmed to be absent from the peripheral blood cells, including the platelets.
Biotin-conjugated goat anti-rabbit IgG heavy and light chains ab, Abcam were reacted against the primary Abs , dilution. Mouse whole blood smears were air-dried and fixed in methanol. Nuclear staining using bisBenzimide H MilliporeSigma was employed where necessary.
Rabbit anti- P. Washing in each step was performed with PBS without any detergent. ELISAs were performed as duplicate assays. DiO fluorescence was detected on the FL-1 channel. The signal distribution of fluorescence shown in Figure 1A that ranges from cell membrane to parasites may suggest a relation to CD36, as CD36 reportedly internalizes holo-particles of HDL Brundert et al.
HDL-uptake by Plasmodium -infected erythrocytes. Phospholipid and ester components were concomitantly observed around parasite, but HDL was not taken up by the nRBCs arrows. Synchronized ring-stage P. D Chemical structure of BLT The BLT-1 effects on parasitized erythrocyte may be due to off-target effects of BLT-1, but it also raises a possibility that other molecules may be influenced by the BLT-1 action in parasitized erythrocytes.
We collected blood from post-infection days 5— Up to day 5, most parasites we observed were ring forms, and very low levels of immunoreactive CD36 were observed. CD36 co-localized with apical membrane antigen 1 AMA-1 Supplementary Figure 3B , a Plasmodium integral membrane protein, suggesting that CD36 may be involved in sterol uptake from erythrocytes to the internalized parasites.
The CD36 signals pink were weak in the pRBCs on days 5 and 8 ring stage , but were abundantly detected on days 11 and 14 schizonts. C Immunocytochemical analysis of pRBCs. CD41 pink signals were absent on days 5 and 8 ring stage , but a small signal emanated from the internal parasites on day 8. CDspecific fluorescence became abundant on days 11 and 14 schizont stage. That CD36 is scarcely distributed on nRBCs provides clues about the specific CD36 translocation pathway for targeting parasitized cells.
Among the hematopoietic cells, platelets and leukocytes particularly those from the monocyte series are known CDpositive cells; thus, CD36 should originate from these cells. CD41 signals also originated from the parasites within Pf -infected erythrocytes. CD41 on each parasite was especially evident in schizonts Supplementary Figure 3E. These data support our hypothesis that molecules from platelets can be transferred to pRBCs, but with an unknown mode of transfer.
Alexandre Siqueira. Francisco De Assis Dias. Gustavo Moreira. Diego Alves. Beatriz Cristina. Rodolfo Afonso. Gustavo Henrique Almeida. Daniela Corado. Lincoln Vinicius. Bianca Farias. Paulo Vicente Correa. Eugenio Manuel. Something went wrong - we were not able to load your agreed MSA pricing. Please try refreshing the page. No reviews have been left for this product. Be the first to leave a review! Looking for support assistance?
Davor Solter Kannagi, R. SSEA-4 can be used as a marker of human embryonic stem cells, human embryonic carcinoma cells and human embryonic germ cells.
Monoclonal antibodies to this target have been widely used in the characterization of pluripotent stem cells. Mouse pluripotent stem cells are not recognised by anti-SSEA-4 antibodies but do express the antigen upon differentiation. Stage specific embryonic antigen 4 antibody. See Abreview. Epub Jul Fig 6. As can be seen from the histograms, in non-differentiating conditions i. However, upon differentiation addition of retinoic acid , the antibody lost the ability to recognise the cells.
As expected, staining localised to the cell surface green. Nuclei are stained blue using Hoechst. Mouse on Mouse staining protocol. Datasheets and documents. Language Select language. References Submit a review Submit a question.
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