Monocyte glycolysis determines CD8+ T cell functionality in human Chagas disease
Liliana María Sanmarco, Natalia Eberhardt, Gastón Bergero, Luz Piedad Quebrada Palacio, Pamela Martino Adami, Laura Marina Visconti, Ángel Ramón Minguez, Yolanda Hernández-Vasquez, Eugenio Antonio Carrera Silva, Laura Morelli, Miriam Postan, Maria Pilar Aoki
Liliana María Sanmarco, Natalia Eberhardt, Gastón Bergero, Luz Piedad Quebrada Palacio, Pamela Martino Adami, Laura Marina Visconti, Ángel Ramón Minguez, Yolanda Hernández-Vasquez, Eugenio Antonio Carrera Silva, Laura Morelli, Miriam Postan, Maria Pilar Aoki
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Research Article Immunology Metabolism

Monocyte glycolysis determines CD8+ T cell functionality in human Chagas disease

Abstract

Chagas disease is a lifelong pathology resulting from Trypanosoma cruzi infection. It represents one of the most frequent causes of heart failure and sudden death in Latin America. Herein, we provide evidence that aerobic glycolytic pathway activation in monocytes drives nitric oxide (NO) production, triggering tyrosine nitration (TN) on CD8+ T cells and dysfunction in patients with chronic Chagas disease. Monocytes from patients exhibited a higher frequency of hypoxia-inducible factor 1α and increased expression of its target genes/proteins. Nonclassical monocytes are expanded in patients’ peripheral blood and represent an important source of NO. Monocytes entail CD8+ T cell surface nitration because both the frequency of nonclassical monocytes and that of NO-producing monocytes positively correlated with the percentage of TN+ lymphocytes. Inhibition of glycolysis in in vitro–infected peripheral blood mononuclear cells decreased the inflammatory properties of monocytes/macrophages, diminishing the frequency of IL-1β– and NO-producing cells. In agreement, glycolysis inhibition reduced the percentage of TN+CD8+ T cells, improving their functionality. Altogether, these results clearly show that glycolysis governs oxidative stress on monocytes and modulates monocyte–T cell interplay in human chronic Chagas disease. Understanding the pathological immune mechanisms that sustain an inflammatory environment in human pathology is key to designing improved therapies.

Authors

Liliana María Sanmarco, Natalia Eberhardt, Gastón Bergero, Luz Piedad Quebrada Palacio, Pamela Martino Adami, Laura Marina Visconti, Ángel Ramón Minguez, Yolanda Hernández-Vasquez, Eugenio Antonio Carrera Silva, Laura Morelli, Miriam Postan, Maria Pilar Aoki

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Figure 4

Monocytes from T.cruzi–infected patients show increased glycolysis and altered mitochondrial potential.

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Monocytes from T.cruzi–infected patients show increased glycolysis and a...
(A) Monocytes from control donors (CON) or patients with Chagas disease (CHAG) isolated by CD14+ selection were seeded in a Seahorse XFp analyzer, and real-time extracellular acidification rate (ECAR) was determined during sequential treatments with glucose, oligomycin (ATP synthase inhibitor) and 2-deoxyglucose (2-DG) (glycolysis inhibitor). Data represent mean ± SEM of 8 runs. (B) Bars show glycolytic rate, glycolytic capacity, and glycolytic reserve of monocytes from CON (n = 5) and CHAG (n = 5). To analyze these parameters, first nonglycolytic acidification was subtracted, and then the data were normalized to basal ECAR. (C) Frequency and MFI of Glut-1+ monocytes from CON (n = 3) and CHAG (n = 5). (D) Real-time mitochondrial respiration was analyzed starting from basal respiration and after the addition of oligomycin (complex V inhibition), FCCP (maximal respiration induction), and rotenone/antimycin A mixture (electron transport chain inhibition). Data represent mean ± SEM of 5 experiments. Bars show (E) respiration driving ATP synthesis, respiration driving proton leak, spare respiratory capacity, nonmitochondrial OCR, and (F) Bioenergetic Health Index (BHI) of monocytes from CON (n = 4) and CHAG (n = 5). To analyze these parameters, first nonmitochondrial OCR was subtracted, and then the data were normalized to basal OCR. (G) Representative histogram of total mitochondrial mass analyzed by flow cytometry in monocytes from CON (black line) and CHAG (gray line) labeled with MitoTracker Green. Mitochondrial membrane potential was analyzed in monocytes from CON (black line) and CHAG (gray line) labeled with MitoTracker Orange. Representative dot plot of mitochondrial mass versus mitochondrial membrane potential in CON (n = 3) and CHAG (n = 5) monocytes. Data are presented as mean ± SEM. *P < 0.05; ***P < 0.001 (Student’s t test or Mann-Whitney U test).