Building a Metabolic Profile for Human Innate-Like T Cells using Spectral Flow Cytometry

Abstract

Obesity is a growing epidemic often leading to metabolic disease, and immune dysregulation is an associated pathology. This dysregulation could be targeted therapeutically to minimise some of the negative effects of obesity, and mitigate pre-diabetes pathogenesis. In this context, innate-like T cells appear to be a particularly suitable target as these T lymphocytes are responsive to their metabolic environment, and are highly enriched in adipose tissue, and are therefore in an ideal position to exert immunometabolic effects on the adipose tissue micro-environment. This project aimed to establish a novel methodology utilising spectral flow cytometry to study the immunometabolic regulation and function of innate-like T cells. Metabolic spectral flow cytometry enables single-cell quantification of functional and metabolic parameters, which is necessary to establish innate-like T cells as viable therapeutic targets, as well as increasing understanding of these cells in general. As access to adipose tissue was limited as a result of the COVID-19 pandemic, this thesis focusses on peripheral blood mononuclear cell (PBMC)-derived cells, and presents the necessary steps to develop a working flow cytometry panel, with the goal of using it to explore innate-like T cell metabolism in both PBMC and adipose tissue-derived cells, from humans with and without metabolic disease. Validation has been conducted on PBMC from 10 healthy participants, with HbA1c measured as an indicator of metabolic health. Preliminary results are presented and discussed.