Protocols for Recinto et al. "A rewiring of the earliest immune events leading to T-cell mediated disease following intestinal microbial infection  in a PINK1 KO mouse model of Parkinson’s disease"

Sherilyn Junelle Recinto; adam.macdonald; Moein Yaqubi; Alexandra Kazanova

Jul 03, 2024

Protocols for Recinto et al. "A rewiring of the earliest immune events leading to T-cell mediated disease following intestinal microbial infection in a PINK1 KO mouse model of Parkinson’s disease"

DOI

dx.doi.org/10.17504/protocols.io.kxygxy77ol8j/v1

¹Department of Neurology and Neurosurgery, Montreal Neurological Institute-Hospital, McGill University, Montréal, QC, Canada;
²Department of Microbiology and Immunology, McGill University, Montréal, QC, Canada

Lilia Rodriguez

Université de Montréal

DOI: dx.doi.org/10.17504/protocols.io.kxygxy77ol8j/v1

Collection Citation: Sherilyn Junelle Recinto, adam.macdonald, Moein Yaqubi, Alexandra Kazanova 2024. Protocols for Recinto et al. "A rewiring of the earliest immune events leading to T-cell mediated disease following intestinal microbial infection in a PINK1 KO mouse model of Parkinson’s disease". protocols.io https://dx.doi.org/10.17504/protocols.io.kxygxy77ol8j/v1

License: This is an open access collection distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited

Protocol status: Working

We use this collection and it's working

Created: June 20, 2024

Last Modified: July 03, 2024

Collection Integer ID: 102180

Keywords: ASAPCRN

Funders Acknowledgement:

Aligning Science Across Parkinson's

Grant ID: ASAP-000525

Abstract

Our group has developed GI-targeted pathogen-induced PD mouse modeling systems (in PINK1 KO mice with gram negative bacterial infections) and found that T cells are a major player in driving PD-like motor symptoms at late stages following infection. Herein, we now map the initiating immune events at the site of infection at the earliest stages with the goal of shedding light on the earliest mechanisms triggering T cell-mediated pathological processes relevant to PD. Using unbiased single cell sequencing, we demonstrate that myeloid cells are the earliest dysregulated immune cell type in PINK1 KO infected mice (at 1-week post-infection) followed by a dysregulated T cell response shortly after (at 2 weeks post-infection). We find that these myeloid cells have an enhanced proinflammatory profile, are more mature, and develop enhanced capacity for antigen presentation. Using unbiased prediction analysis, our data suggests that cytotoxic T cells and myeloid cells are particularly poised for interacting with each other, and we identify possible direct cell-cell interaction pathways that might be implicated. 

Files

SR2024_Immunophenotyping_flowcytometry.docx

SR2024_gutLP_prep.docx

SR2023_LipocalinELISA.docx

SR2022_10XChromium single cell 3'RNAseq preparation.docx

SR2024_RNAextraction+qPCR.docx

SR2024_Immunophenotyping_flowcytometry.docx

Procedure_Citrobacter rodentium infection model SOP.docx

Public workspaceProtocols for Recinto et al. "A rewiring of the earliest immune events leading to T-cell mediated disease following intestinal microbial infection in a PINK1 KO mouse model of Parkinson’s disease"

Protocols for Recinto et al. "A rewiring of the earliest immune events leading to T-cell mediated disease following intestinal microbial infection in a PINK1 KO mouse model of Parkinson’s disease"