Mitochondrial Antigen Presentation (MitAP) to 2CZ CD8+ T cell hybridoma

By Alexandra Kazanova, PhD
View Published Protocol

Output Details

Description
This protocol details the Mitochondrial Antigen Presentation (MitAP) to 2CZ CD8+ T cell hybridoma. Previously, quantification of the number of IL-2 producing 2cz hybridoma cells in an ELISPOT assay was used as a readout for MitAP (Matheoud et al., 2016). Here, we adapted the assay for the detection of activation-induced markers (AIM) on 2cz cells with flow cytometry.
Identifier (DOI)
10.17504/protocols.io.5qpvok9j7l4o/v1
Tags
  • Activity Assay
  • Antigen-presenting cells (APCs)
  • Antigen-specific response
  • Mitochondrial
Team
Team Desjardins
Program
Collaborative Research Network
Theme
Neuro-Immune Interactions

Meet the Authors

  • Alexandra Kazanova, PhD

    Key Personnel: Team Desjardins, 

    McGill University

Related Research

Modeling gene-environment interactions in Parkinson’s Disease: Helicobacter pylori infection of Pink1-/- mice induces CD8 T cell-dependent motor and cognitive dysfunction.

Here, we demonstrate in a mouse model deficient in the PD-associated gene Pink, that infection with the human PD-associated gastric bacterium Helicobacter pylori leads to development of motor and cognitive signs resembling prodromal features of PD.

View Article

Lysosomal GCase (glucocerebrosidase) activity assay

Method to measure lysosomal GBA1 enzyme activity using 4-MUG substrate at low pH to minimize non-lysosomal GCase interference. GBA1 activity higher in purified lysosomes than whole cell extract, confirmed by CBE inhibitor abolishing 4-MUG hydrolysis.

View Protocol

Ex Vivo Mouse Brain Slice Whole-Cell Patch-Clamp Electrophysiology

This protocol outlines steps for performing ex vivo slice whole-cell patch-clamp electrophysiology experiments, including voltage clamp and current clamp recordings as well as optogenetic stimulation in the Ding lab.

View Protocol
View More Outputs