A Human Blood-Brain Interface Model to Study Barrier Crossings by Pathogens or Medicines and Their Interactions with the Brain

作者： Anaelle da Costa*1, Christophe Prehaud*1, Florian Bakoa1, Philippe Afonso2, Pierre-Emmanuel Ceccaldi2, Pierre Lafaye3, Monique Lafon1 1Institut Pasteur, CNRS UMR 3569, Unité de Neuroimmunologie Virale, 2Institut Pasteur, Unité d’Epidémiologie et de Pathophysiologie des Virus Oncogènes, Université Sorbonne Paris Cité/Paris Diderot, 3Institut Pasteur, PFIA, DTPS

简介：

Overview

This protocol outlines the establishment of a human blood-brain barrier (BBB) minibrain model using polyester porous membrane culture inserts to investigate the transport of biomolecules and infectious agents into the central nervous system. The study aims to evaluate the physiological effects on adjacent brain cells, providing insights into the interactions between the BBB and various pathogens.

Key Study Components

Area of Science

• Neuroscience
• Cell Biology
• Infectious Disease

Background

• The blood-brain barrier is vital for protecting the brain while also posing challenges for drug delivery and pathogen access.
• The minibrain model facilitates interactions between human neurons and the BBB.
• This approach allows for assessing permeability and physiological impacts of various molecules.

Purpose of Study

• To evaluate the transport of biomolecules and viruses across the BBB.
• To understand the effects of pathogens on neuronal cells.
• To develop a reliable model for future drug testing and neurobiological studies.

Methods Used

• The study utilized a cell culture system involving polyester membrane inserts within a 12-well plate.
• The biological model included human cerebral microvascular endothelial cells in conjunction with human neurons and astrocytes.
• Critical steps included cell cultivation to confluency, application of viral agents, and measurement of endothelial permeability.
• The set-up was incubated, and various assays were performed at specified intervals to evaluate results.

Main Results

• The study confirmed that certain viruses can cross the BBB and multiply within brain cells.
• Cell-penetrating molecules demonstrated the ability to regenerate axons and promote neuroprotection.
• Validation experiments showed a direct correlation between viral load and specific biomarker expression.

Conclusions

• This study enables enhanced understanding of BBB dynamics and interactions with pathogens.
• The findings support the development of therapeutic strategies targeting the neurovascular interface.
• Implications extend to potential interventions for diseases affecting the brain.

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