Quick Project Snapshot
Do circulating microvesicles from patients with multiple sclerosis (MS) disrupt the blood-brain barrier (BBB)? (For Honors and MBiomedSc)
Breakdown of the blood brain barrier (BBB) precedes clinical symptoms of new lesions of MS and it is possible that high numbers of microvesicles in multiple sclerosis (MS) plasma are related to episodes of disruption of the BBB. The integrity of BBB will be studied using an in vitro model examining lymphocyte transmigration across confluent monolayers of cultured endothelial cells. Human umbilical vein endothelial cells (HUVECs) are grown to confluent monolayers in tissue culture plates and peripheral blood lymphocytes added to each well and incubated for 2-4 h. The HUVEC layer is washed 5 times with saline media, then fixed and examined by phase-contrast microscopy. Cells beneath the monolayer appear phase dark while adherent cells above appear phase light. The number of adherent and migrated cells are counted to give an index of efficiency of migration. To assess if microvesicles impair the integrity of the endothelial monolayer, the migration assay will be performed both in the absence and presence of plasma containing known concentrations of platelet derived microvesicles. Meanwhile, the lysosomal β-hesosaminidase activity will be measured in platelet poor plasma from 20 MS patients and 20 controls using a standard colourimetric assay. The microvesicle counts, β-hesosaminidase activity and the impact on lymphocytes transendothelial migration will be analysed in correlation. Results could provide evidence for a mechanism by which peripheral blood leukocytes infiltrate to brain in MS.
Techniques involved include cell culture, ultra-centrifugation, flow cytometry, fluorescent microscopy and biochemistry.
Innate Phagocytosis Laboratory
The Innate Phagocytosis Laboratory led by Dr. Ben J. Gu and Prof. James S. Wiley has great interests in discovering the role of innate phagocytosis in neurodegeneration, neurodevelopment, infection and cancer. This Lab has a long history of studies on the P2X7 receptor, an ion channel activated by ATP. They discovered the non-functional P2X7 and identified a number of genetic variations affecting P2X7 pore formation and pro-inflammatory responses. Recently they have found that P2X7 is in fact a scavenger receptor in the absence of ATP or serum. Further study of P2X7 mediated innate phagocytosis opens a new research field for its role in broad biological aspects, including:
- Neurodegeneration: This lab explores genetic, cell biological and animal studies to identify a common mechanism underlying a number of neurological disorders including Alzheimer’s disease, age-related macular degeneration and multiple sclerosis.
- Neurodevelopment: This lab has revealed that P2X7 mediated phagocytosis probably contributes to the early development of central nervous system by removing apoptotic neuronal cells.
- Infection: Innate immunity can be promoted by neutralizing certain glycoprotein in serum or by compounds promoting innate phagocytosis. This lab is currently developing a rabbit model of bacteraemia/sepsis.
- Cancer: The non-functional P2X7 is found to be expressed on the surface of various tumor cells, but not on normal cells. The antibody against this non-functional P2X7 receptor is currently undergoing a clinical trial for basal cell carcinoma conducted by Biosceptre Pty Ltd. This lab has great interests in investigating the biological nature of non-functional P2X7, P2X7 mediated phagocytosis and oncogenesis.
All Projects by this LabSearch the P2X7 related biomarkers for Alzheimer’s diseaseThe nature of the P2X4 receptorThe role of P2X7 in a mouse model of oligodendrocyte apoptosisInvestigation of P2X7 mediated phagocytosis - potential therapeutic target for cancer treatmentIdentify the transcriptional regulatory factors of the P2X7 receptorHow does the brain remove the excess number of neurons during development and agingIdentification of serum glycoproteins altering innate immunityDo circulating microvesicles from patients with multiple sclerosis (MS) disrupt the blood-brain barrier (BBB)? (For Honors and MBiomedSc)The Role of Innate Phagocytosis in the Pathogenesis and Treatment of Alzheimer's Disease (for PhD only)
The Florey's Epilepsy division is a world-leading centre for epilepsy research. The division has major groups at both the Florey’s Austin and Parkville campus. The group studies mechanisms that cause epilepsy from the level of cells to the function of the whole brain. We use technologies including advanced MRI and cutting edge cellular physiology techniques to allow us to understand genetic and acquired mechanisms that give rise to epilepsy. Together with our colleagues from The University of Melbourne and across Australia we are working towards finding a cure for epilepsy.