HIF1A-MEDIATED GLYCOLYSIS PROMOTES THE MIGRATORY CAPACITY OF DENDRITIC CELLS IN TUBERCULOSIS

MARIANO MAIO; JOSÉ LUIS MARÍN FRANCO; ROSA MUSELLA; GEANNCARLO LUGO VILLARINO; CHRISTEL VEROLLET; ZOI VAHLAS; MELANIE GENOULA; LORENA CIALLELLA; OLIVIER NEYROLLES; LUCIANA BALBOA; JOAQUINA BARROS; FEDERICO FUENTES; DOMINGO PALMERO; MARÍA DEL CARMEN SASIAIN

sao paulo

Congreso; XLVI congress of the brazilian society of immunology; 2022

IntroductionMycobacterium tuberculosis (Mtb) is a highly successful pathogen that interferes with dendritic cells (DCs) functions impairing the onset and development of adaptive immunity, favoring Tuberculosis (TB). Since several studies are now revealing the importance of metabolic pathways involved in DCs functions, we wondered how the metabolic pathways influence DCs activation in response to Mtb. MethodsBuffy coats from healthy donors were prepared at the Garrahan Hospital (Buenos Aires), according to institutional guidelines (CEIANM-664/07). Peripheral blood samples from patients with TB were provided by the Muñiz Hospital. Written informed consent was obtained in accordance with the guidelines of the hospital´s ethics committee (protocol number: 2110/21). Monocyte-derived DCs were stimulated with equivalent doses of either irradiated Mtb or viable Mtb. After 24h, the metabolic profile of DCs was determined by Seahorse and SCENITH technologies, lactate release and glucose consumption measured by colorimetric assays and HIF-1α and LDHA expression by qPCR and flow cytometry. Also, mitochondrial morphology was determined by transmission electron microscopy. When indicated blocking antibodies against TLR2 or TLR4 were added. Besides, HIF-1α activity was modulated by using either PX-478, a HIF-1α inhibitor, or DMOG, a HIF-1α activator. To assess the participation of glycolysis, the drug GSK28378 or sodium oxamate were used. Finally, the capacity of DCs to migrate was evaluated by chemotactic assays towards CCL21 and 3D migration through fibrillar collagen. ResultsMtb-stimulated DCs displayed a predominant glycolytic profile in a TLR2-dependent manner. Additionally, Mtb-triggered HIF-1α-mediated glycolysis was required for DCs to adopt a migratory phenotype, since either HIF-1α activity or glycolysis inhibition abolished the trafficking properties. Besides, the stabilization of HIF-1α promoted the migration towards CCL21 and the 3D-migration of cells throughout the collagen matrix in tolerogenic DCs induced by dexamethasone. Finally, the stabilization of HIF-1α promotes the chemotactic activity in DCs derived from TB patients. ConclusionsOur data provide novel insights into immunometabolic pathways involved in the trafficking of Mtb-activated DCs to the lymph nodes that could be targeted to generate sterilizing vaccine-induced immunity against TB.