Modelling T-cell immunity against hepatitis C virus with liver organoids in a microfluidic coculture system

Area of Science:

• Immunology
• Hepatology
• Biotechnology

Background:

• Hepatitis C virus (HCV) infection affects millions globally, necessitating better research models due to limited vaccines and animal models.
• Existing in vitro models often exhibit aberrant immune and proliferative responses, hindering accurate study of the human immune system's interaction with HCV.
• Studying the adaptive immune response to HCV requires advanced in vitro systems that utilize primary human cells.

Purpose of the Study:

• To develop and validate a novel microfluidic coculture system for studying human adaptive immune responses to Hepatitis C virus (HCV).
• To enable molecular investigation of T cell-mediated immunity against HCV using primary human cells in a controlled in vitro environment.

Main Methods:

• A microfluidic chip was used to coculture 3D human liver organoids with primary human CD8+ T cells.
• Organoids were embedded in extracellular matrix, and T cells were cultured in suspension, maintaining HLA matching.
• Automated phase contrast and immunofluorescence imaging were utilized to observe T cell invasion and organoid morphology.

Main Results:

• The microfluidic coculture system successfully supported T cell-mediated killing of liver organoids.
• Targeted killing was observed when organoids were exposed to a peptide specific for HCV non-structural protein 3 (NS3) and cognate CD8+ T cells.
• The system demonstrated the capacity to study adaptive immune responses to HCV in vitro.

Conclusions:

• The developed microfluidic coculture system provides a novel platform for investigating adaptive immune responses to HCV using primary human cells.
• This system offers a valuable tool for molecular studies of T cell interactions with liver cells in the context of HCV infection.
• The model holds potential for advancing research into HCV immunity and developing new therapeutic strategies.