Measuring ATP Hydrolysis in T Cell Subsets

Sorted CD39⁺ Tconv, CD39⁻ Tconv and Treg cells from B16F10-OVA tumor-bearing Foxp3-EGFP mice were incubated with 5 µM of exogenous ATP (Sigma-Aldrich) as previously described.^{19 (link)} Cell-free medium was then analyzed for residual ATP with ATP bioluminescent assay kit (INVITROGEN). Bioluminescent activity was measured with a Synergy HTX Multi-Mode Reader.

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Bossio S.N., Abrate C., Tosello Boari J., Rodriguez C., Canale F.P., Ramello M.C., Brunotto V., Richer W., Rocha D., Sedlik C., Vincent-Salomon A., Borcoman E., Del Castillo A., Gruppi A., Fernandez E., Acosta Rodríguez E.V., Piaggio E, & Montes C.L. (2023). CD39+ conventional CD4+ T cells with exhaustion traits and cytotoxic potential infiltrate tumors and expand upon CTLA-4 blockade. Oncoimmunology, 12(1), 2246319.

Publication 2023

exogenous ATP ATP bioluminescent assay kit Synergy HTX Multi-Mode Reader

Corresponding Organization :

Other organizations : Research Centre in Biological Chemistry of Córdoba, Consejo Nacional de Investigaciones Científicas y Técnicas, Universidad Nacional de Córdoba, Université Paris Sciences et Lettres, Inserm, Institut Curie

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Variable analysis

independent variables

Incubation of sorted CD39+ Tconv, CD39- Tconv, and Treg cells from B16F10-OVA tumor-bearing Foxp3-EGFP mice with 5 µM of exogenous ATP (Sigma-Aldrich)

dependent variables

Residual ATP in the cell-free medium, measured using an ATP bioluminescent assay kit (INVITROGEN)

control variables

Incubation conditions (e.g., temperature, duration)
Assay kit and equipment (Synergy HTX Multi-Mode Reader)

controls

Positive control: None specified
Negative control: None specified

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