Using spectral flow cytometry and BD Horizon RealYellow™ and BD Horizon RealBlue™ Reagents to dissect the expression of metabolic proteins in resting and activated T cells
Year: 2023
Type: conference-abstract
Abstract: In response to activation, T cells undergo metabolic adaptations to meet the energetic and biosynthetic demands required for their rapid cell growth and proliferation. T cell receptor (TCR) and CD28 costimulation trigger the initial metabolic changes that support T cell activation including increase in glucose uptake. PI3K signaling is critical for upregulation of Glut1, the main glucose receptor in T cells. In this study, we describe an assay for detection of Glut1 and key metabolic enzymes using fluorochrome-conjugated antibodies that have been validated for flow cytometry. Our analyses showed a broad expression of Glut1 and other metabolic markers across human immune cell populations. In T cells, Glut1 was expressed in resting T cells and upregulated in response to anti-CD3/CD28 stimulation. Glut-1high cells were found among CD69+ activated T cells, which also showed higher expression levels of glucose-6-phosphate dehydrogenase (G6PD) compared to the CD69− cells. Notably, the PI3K inhibitor Ly294002 abrogated the upregulation of Glut1 and G6PD expression in activated CD69+ T cells. Ly294002 cell treatment did not affect the expression of hexokinase 1 (HK1), CD69, PD-1 or CD25, suggesting that PI3K inhibition impacted the expression of select metabolic markers and not overall T cell activation. These results support literature evidence showing the importance of the glycolytic pathway on T cell activation while demonstrating optimized experimental conditions for resolution of multiple metabolic markers. For Research Use Only. Not for use in diagnostic or therapeutic procedures. BD, BD Horizon RealBlue and BD Horizon Real Yellow are trademarks of Becton, Dickinson and Company or its affiliates. © 2023 BD. All rights reserved.
Source: The Journal of Immunology
Language: English
Topic: T-cell and B-cell Immunology
Subfield: Immunology
Field: Immunology and Microbiology
Domain: Life Sciences
Open Access status: bronze
OpenAlex ID: W4385687185