T cell responses towards PINK1 and ⍺-synuclein are elevated in prodromal Parkinson's disease - Fluorospot data

Antigen-specific T cell reactivity in PBMCs were studied using the Fluorospot assay

PBMCs were co-cultured with alpha-synuclein, (⍺-syn) PINK1, or Epstein-Barr virus peptide pools and subsequently expanded in vitro over a two-week period. Cultures were supplemented with IL-2 (10 U/mL) every 4 days. After two weeks, expanded cells were harvested and were re-stimulated in triplicates with the same peptide pools, dimethylsulfoxide (DMSO; as a negative control) and phytohemagglutinin (PHA; as a positive control) and then processed for a dual-color (interferon-gamma and IL-5) Fluorospot assay (Mabtech). Spot forming cells (SFC) were captured using an IRIS Fluorospot reader (Mabtech) and results were analyzed using Excel (Microsoft). To be considered positive, antigen-specific T cells had >100 SFC per million PBMC, p-value <0.05 and >2 fold increase when compared to the DMSO control.

Note: “ND” refers to “no data”, as some samples either did not have the adequate amount of cells for testing or did not pass our SFC quality control criteria. The "Sum of ⍺-syn and PINK1 IFNγ SFC/10^6 cultured cells" and "Sum of ⍺-syn and PINK1 IL-5 SFC/10^6 cultured cells" was only calculated for donors screen with both PINK1 and ⍺-syn.

In the Unified Parkinson’s Disease Rating Scale (UPDRS) part III and the Montreal Cognitive Assessment (MoCA) tests columns, donors not tested were assigned "Not tested".