Abstract
In this work, the effects of the protozoan Neospora caninum on the bioenergetics, chemical composition, and elemental content of human brain microvascular endothelial cells (hBMECs) were investigated. We showed that N. caninum can impair cell mitochondrial (Mt) function and causes an arrest in host cell cycling at S and G2 phases. These adverse effects were also associated with altered expression of genes involved in Mt energy metabolism, suggesting Mt dysfunction caused by N. caninum infection. Fourier Transform Infrared (FTIR) spectroscopy analysis of hBMECs revealed alterations in the FTIR bands as a function of infection, where infected cells showed alterations in the absorption bands of lipid (2924 cm−1), amide I protein (1649 cm−1), amide II protein (1537 cm−1), nucleic acids and carbohydrates (1092 cm−1, 1047 cm−1, and 939 cm−1). By using quantitative synchrotron radiation X-ray fluorescence (μSR-XRF) imaging and quantification of the trace elements Zn, Cu and Fe, we detected an increase in the levels of Zn and Cu from 3 to 24 h post infection (hpi) in infected cells compared to control cells, but there were no changes in the level of Fe. We also used Affymetrix array technology to investigate the global alteration in gene expression of hBMECs and rat brain microvascular endothelial cells (rBMVECs) in response to N. caninum infection at 24 hpi. The result of transcriptome profiling identified differentially expressed genes involved mainly in immune response, lipid metabolism and apoptosis. These data further our understanding of the molecular events that shape the interaction between N. caninum and blood-brain-barrier endothelial cells.
Highlights
Neospora caninum is an obligate intracellular apicomplexan protozoan parasite
We presented our attempt to correlate the findings obtained from investigation of the viability, bioenergetic, chemical signature, elemental profile, and gene expression of blood brain barrier (BBB) endothelial cells infected by N. caninum
The Fourier Transform Infrared (FTIR) spectroscopic fingerprinting analysis of human brain microvascular endothelial cells (hBMECs) at 3, 24 and 48 hpi showed that N. caninum infection caused significant changes in the chemical composition of infected cells
Summary
Neospora caninum is an obligate intracellular apicomplexan protozoan parasite. An association of this parasite with abortion in cattle [1,2] and neuromuscular disease in dogs has been established based on seroepidemiological and pathological studies [2,3,4,5,6]. The BBB is an active tissue made up of astrocytes, pericytes and microvascular endothelial cells, and selectively controls intracellular and paracellular passage of substances between the CNS and blood [9,10]. These cellular components along with closely packed neurons constitute the neurovascular component, which forms the major functional unit of the BBB [9]. Understanding the molecular changes that occur in the microvascular endothelial cells due to N. caninum infection is important because these cells maintain the functional integrity of the BBB and provide a highly selective barrier that protects the brain against pathogen invasion. Earlier studies have revealed disruption of the bioenergetics of microvascular endothelial cells in response to N. caninum infection [11,12]
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