A rapid and scalable density gradient purification method for Plasmodium sporozoites

Abstract Background Malaria remains a major human health problem, with no licensed vaccine currently available. Malaria infections initiate when infectious Plasmodium sporozoites are transmitted by Anopheline mosquitoes during their blood meal. Investigations of the malaria sporozoite are, therefore...

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Bibliographic Details
Published in:Malaria Journal
Main Authors: Kennedy Mark, Fishbaugher Matthew E, Vaughan Ashley M, Patrapuvich Rapatbhorn, Boonhok Rachasak, Yimamnuaychok Narathatai, Rezakhani Nastaran, Metzger Peter, Ponpuak Marisa, Sattabongkot Jetsumon, Kappe Stefan H, Hume Jen CC, Lindner Scott E
Format: Article in Journal/Newspaper
Language:English
Published: BMC 2012
Subjects:
Sporozoite
Purification
Plasmodium
Falciparum
Vivax
Yoelii
Humanized mouse model
Arctic medicine. Tropical medicine
RC955-962
Infectious and parasitic diseases
RC109-216
Arctic
Human health
Online Access:https://doi.org/10.1186/1475-2875-11-421
https://doaj.org/article/7212a77b5b3e4075b0032434e327a99c
Description
Summary:Abstract Background Malaria remains a major human health problem, with no licensed vaccine currently available. Malaria infections initiate when infectious Plasmodium sporozoites are transmitted by Anopheline mosquitoes during their blood meal. Investigations of the malaria sporozoite are, therefore, of clear medical importance. However, sporozoites can only be produced in and isolated from mosquitoes, and their isolation results in large amounts of accompanying mosquito debris and contaminating microbes. Methods Here is described a discontinuous density gradient purification method for Plasmodium sporozoites that maintains parasite infectivity in vitro and in vivo and greatly reduces mosquito and microbial contaminants. Results This method provides clear advantages over previous approaches: it is rapid, requires no serum components, and can be scaled to purify >10 7 sporozoites with minimal operator involvement. Moreover, it can be effectively applied to both human ( Plasmodium falciparum , Plasmodium vivax ) and rodent ( Plasmodium yoelii ) infective species with excellent recovery rates. Conclusions This novel method effectively purifies viable malaria sporozoites by greatly reducing contaminating mosquito debris and microbial burdens associated with parasite isolation. Large-scale preparations of purified sporozoites will allow for enhanced in vitro infections, proteomics, and biochemical characterizations. In conjunction with aseptic mosquito rearing techniques, this purification technique will also support production of live attenuated sporozoites for vaccination.

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