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Isolation and Analysis of Brain-sequestered Leukocytes from Plasmodium berghei ANKA-infected Mice
Published on: January 2, 2013
Investigation of Plasma-Derived Lipidome Profiles in Experimental Cerebral Malaria in a Mouse Model Study
Amani M Batarseh1,2, Fatemeh Vafaee3,4,5, Elham Hosseini-Beheshti6
1Sydney Knowledge Hub, BCAL Dx Ltd., The University of Sydney, Merewether Building, Sydney, NSW 2006, Australia.
Abstract:
Cerebral malaria (CM), a fatal complication of Plasmodium infection that affects children, especially under the age of five, in sub-Saharan Africa and adults in South-East Asia, results from incompletely understood pathogenetic mechanisms. Increased release of circulating miRNA, proteins, lipids and extracellular vesicles has been found in CM patients and experimental mouse models. We compared lipid profiles derived from the plasma of CBA mice infected with Plasmodium berghei ANKA (PbA), which causes CM, to those from Plasmodium yoelii (Py), which does not. We previously showed that platelet-free plasma (18k fractions enriched from plasma) contains a high number of extracellular vesicles (EVs). Here, we found that this fraction produced at the time of CM differed dramatically from those of non-CM mice, despite identical levels of parasitaemia. Using high-resolution liquid chromatography-mass spectrometry (LCMS), we identified over 300 lipid species within 12 lipid classes. We identified 45 and 75 lipid species, mostly including glycerolipids and phospholipids, with significantly altered concentrations in PbA-infected mice compared to Py-infected and uninfected mice, respectively. Total lysophosphatidylethanolamine (LPE) levels were significantly lower in PbA infection compared to Py infection and controls. These results suggest that experimental CM could be characterised by specific changes in the lipid composition of the 18k fraction containing circulating EVs and can be considered an appropriate model to study the role of lipids in the pathophysiology of CM.
Insights
Cerebral malaria (CM) is linked to distinct lipid changes in circulating extracellular vesicles (EVs). This study identifies specific lipid alterations in experimental CM, offering a new model for understanding disease pathophysiology.
Area of Science:
- Biochemistry
- Immunology
- Pathophysiology
Background:
- Cerebral malaria (CM) is a severe complication of Plasmodium infection with poorly understood mechanisms.
- Increased extracellular vesicles (EVs) and their cargo (miRNAs, proteins, lipids) are observed in CM patients and models.
- Platelet-free plasma fractions (18k) are enriched in EVs and reflect disease state.
Purpose of the Study:
- To compare lipid profiles in plasma EVs from mice infected with CM-inducing Plasmodium berghei ANKA (PbA) versus non-CM-inducing Plasmodium yoelii (Py).
- To identify specific lipid alterations associated with experimental CM.
- To evaluate the 18k plasma EV fraction as a model for studying CM lipid pathophysiology.
Main Methods:
- Infection of CBA mice with PbA (causes CM) or Py (does not cause CM).
- Isolation of platelet-free plasma 18k fractions enriched in EVs.
- High-resolution liquid chromatography-mass spectrometry (LCMS) for lipidomic analysis.
- Comparison of lipid profiles between PbA-infected, Py-infected, and uninfected mice.
Main Results:
- The 18k EV-enriched plasma fraction composition differed significantly between CM and non-CM mice, despite similar parasitemia.
- LCMS identified over 300 lipid species; 45 and 75 species showed altered concentrations in PbA-infected mice compared to Py-infected and uninfected controls, respectively.
- Specific lipid classes, including glycerolipids and phospholipids, were significantly altered.
- Total lysophosphatidylethanolamine (LPE) levels were significantly lower in PbA infection.
Conclusions:
- Experimental CM is characterized by specific changes in the lipid composition of circulating EVs.
- The 18k plasma EV fraction is a suitable model for investigating the role of lipids in CM pathogenesis.
- Lipidomic profiling of EVs may provide insights into CM mechanisms and potential biomarkers.
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