Unravelling malaria parasite rhythms : the development of a flow cytometric assay to quantify and stage Plasmodium Chabaudi parasites

Deodatus, A. (Arne) (2017) Unravelling malaria parasite rhythms : the development of a flow cytometric assay to quantify and stage Plasmodium Chabaudi parasites. thesis, Medicine.

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Abstract

BACKGROUND – Malaria, caused by Plasmodium parasites, remains a primary cause of death in endemic areas. Plasmodium replicates asexually in the blood and parasites within each replication cycle develop synchronously. Disrupting replication rhythms is costly to parasites, opening up novel approaches for treatment. First, a better understanding of parasite rhythms is required but current methods to study rhythms are time consuming and subjective. We aimed to develop a flow cytometric method to overcome this. METHODS – We used dyes (Hoechst 33342) and fluorescently labelled P. chabaudi parasites expressing GFP or mCherry to identify and stage infected red blood cells. We compared flow cytometry and microscopy, and used fluorescence activated cell sorting to investigate developmental stage markers. Finally, we evaluated the new method in an experiment testing how parasite rhythms are established. RESULTS – Microscopic and flow cytometric parasite counts correlated closely, and GFP and mCherry fluorescence are effective parasite stage markers when incorporated in the parasite genome, whereas Hoechst is not. The method can quantify and stage infections of two different parasite strains within the same host. Furthermore, parasites with mismatched rhythms change the length of their asexual cycle to match their rhythm to the host. CONCLUSION – We provide a powerful new flow cytometric method to quantify parasite dynamics and rhythms in a number of scenarios. Implementing the methods revealed plasticity in the duration of the asexual cycle that allows parasites to most efficiently adjust their timing to that of the host.

Item Type:	Thesis (Thesis)
Supervisor name:	faculty Supervisor: and Lisman, Prof. Ton
Supervisor name:	Local Supervisor: and Reece, Prof. Sarah
Faculty:	Medical Sciences
Date Deposited:	25 Jun 2020 10:41
Last Modified:	25 Jun 2020 10:41
URI:	https://umcg.studenttheses.ub.rug.nl/id/eprint/252

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