1. PfCap380 as a marker for Plasmodium falciparum oocyst development in vivo and in vitro
- Author
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Samrita Dungel, Yaxian Zhou, Leslie S. Itsara, Ashley M. Vaughan, Anil K. Ghosh, Stefan H. I. Kappe, Matthew Fishbaugher, Julie Do, Will Betz, Thao Nguyen, Mary Jane Navarro, and Erika L. Flannery
- Subjects
Genetic Markers ,0301 basic medicine ,lcsh:Arctic medicine. Tropical medicine ,lcsh:RC955-962 ,animal diseases ,030231 tropical medicine ,Plasmodium falciparum ,Microbiology ,lcsh:Infectious and parasitic diseases ,03 medical and health sciences ,Oocyst ,0302 clinical medicine ,Malaria vaccines ,Limit of Detection ,In vivo ,parasitic diseases ,Gametocyte ,Humans ,Plasmodium berghei ,lcsh:RC109-216 ,Malaria, Falciparum ,Ookinete ,Anopheles stephensi ,Capsule ,biology ,Research ,fungi ,Oocysts ,DNA, Protozoan ,Culturing system ,biology.organism_classification ,In vitro ,Molecular Typing ,Circumsporozoite protein ,030104 developmental biology ,Infectious Diseases ,PfCap380 ,Polyclonal antibodies ,biology.protein ,Parasitology - Abstract
Background Despite the importance of the Plasmodium berghei oocyst capsule protein (PbCap380) in parasite survival, very little is known about the orthologous Plasmodium falciparum capsule protein (PfCap380). The goal of this work was to study the growth of P. falciparum oocysts using PfCap380 as a developmental marker. Methods To study P. falciparum oocyst development using both in vivo (mosquito-derived) and in vitro (culture-derived) growth conditions, antibodies (polyclonal antisera) were raised against PfCap380. For studies on in vivo oocysts, mature P. falciparum gametocytes were fed to Anopheles stephensi mosquitoes. For studies on in vitro parasites, P. falciparum gametocytes were induced and matured for subsequent ookinete production. Ookinetes were purified and then tested for binding affinity to basal lamina components and transformation into early oocysts, which were grown on reconstituted basal lamia coated wells with novel oocyst media. To monitor in vivo oocyst development, immunofluorescence assays (IFA) were performed using anti-PfCap380 antisera on Pf-infected mosquito midguts. IFA were also performed on culture-derived oocysts to follow in vitro oocyst development. Results The anti-PfCap380 antisera allowed detection of early midgut oocysts starting at 2 days after gametocyte infection, while circumsporozoite protein was definitively observed on day 6. For in vitro culture, significant transformation of gametocytes to ookinetes (24%) and of ookinetes to early oocysts (85%) was observed. After screening several basal lamina components, collagen IV provided greatest binding of ookinetes and transformation into early oocysts. Finally, PfCap380 expression was observed on the surface of culture-derived oocysts but not on gametocytes or ookinetes. Conclusions This study presents developmental monitoring of P. falciparum oocysts produced in vivo and in vitro. The anti-PfCap380 antisera serves as an important reagent for developmental studies of oocysts from the mosquito midgut and also from oocyst culture using in vitro methodology. The present data demonstrate that PfCap380 is a useful marker to follow the development and maturation of in vivo and in vitro produced oocysts as early as 2 days after zygote formation. Further in vitro studies focused on oocyst and sporozoite maturation will support the manufacturing of whole sporozoites for malaria vaccines. Electronic supplementary material The online version of this article (10.1186/s12936-018-2277-6) contains supplementary material, which is available to authorized users.
- Published
- 2018
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