PI3K–Akt signaling controls PFKFB3 expression during human T-lymphocyte activation
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Lymphocyte activation is associated with rapid increase of both the glycolytic activator fructose 2,6-bisphosphate (Fru-2,6-P2) and the enzyme responsible for its synthesis, 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase (PFK-2/FBPase-2). PFKFB3 gene, which encodes for the most abundant PFK-2 isoenzyme in proliferating tissues, has been found overexpressed during cell activation in several models, including immune cells. However, there is limited knowledge on the pathways underlying PFKFB3 regulation in human T-lymphocytes, and the role of this gene in human immune response. The aim of this work is to elucidate the molecular mechanisms of PFKFB3 induction during human T-lymphocyte activation by mitotic agents. The results obtained showed PFKFB3 induction during human T-lymphocyte activation by mitogens such as phytohemagglutinin (PHA). PFKFB3 increase occurred concomitantly with GLUT-1, HK-II, and PCNA upregulation, showing that mitotic agents induce a metabolic reprograming process that is required for T-cell proliferation. PI3K–Akt pathway inhibitors, Akti-1/2 and LY294002, reduced PFKFB3 gene induction by PHA, as well as Fru-2,6-P2 and lactate production. Moreover, both inhibitors blocked activation and proliferation in response to PHA, showing the importance of PI3K/Akt signaling pathway in the antigen response of T-lymphocytes. These results provide a link between metabolism and T-cell antigen receptor signaling in human lymphocyte biology that can help to better understand the importance of modulating both pathways to target complex diseases involving the activation of the immune system.
KeywordsPFKFB3 PI3K–Akt Mitogens Lymphocytes Glycolysis Metabolism
Activation-induced cell death
Carboxyfluorescein succinimidyl ester
Fetal bovine serum
Glucose transporter 1
Interleukin-2 receptor alpha chain
Mean fluorescence intensity
Mammalian target of rapamycin
Mammalian target of rapamycin complex 1
Peripheral blood mononuclear cells
Proliferating cell nuclear antigen
S6 ribosomal protein
T-cell antigen receptor
We are grateful to B. Torrejón from the Microscopy Unit of Centres Científics i Tecnològics of the Universitat de Barcelona (CCiT-UB) for excellent technical assistance and advice; A. M. Cosialls for technical advice in blood cells cultures; J. L. Rosa and F. Viñals for providing reagents; P. Giménez-Bonafé for English revision and E. Adanero for technical assistance.
RB and AM conceived and designed the experiments; HS, CA, PF, and AV performed the experiments; HS, CA, AM, PF, AV, and EC analyzed the data; NLL, AR, and AN critically commented and revised the work; and HS, RB, and AM wrote the manuscript.
This work was supported by Instituto de Salud Carlos III—FIS [PI13/0096] and FIS [PI17/00412]—and Fondo Europeo de Desarrollo Regional (FEDER), and by Astellas European Foundation Award (13th European Society of Transplantation). HS was recipient of a fellowship from Secretaria d’Universitats i Recerca del Departament d’Economia i Coneixement de la Generalitat de Catalunya and AR was recipient of a fellowship from the University of Barcelona.
Compliance with ethical standards
Conflict of interest
The authors declare no conflict of interest.
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