Extracellular vesicles released by triiodothyronine-stimulated dendritic cells induce a pro-inflammatory profile of allogenic murine splenocytes

SOLER MARÍA FLORENCIA; DONADIO ANA CAROLINA; ALAMINO VANINA ALEJANDRA; BLANCO ANTONELLA; CLAUDIA GABRIELA PELLIZAS; NEGRETTI BORGA DANA; BRAVO MIANA ROCÍO; MARIA DEL MAR MONTESINOS

Buenos Aires. virtual

Congreso; 19th International Congress of Endocrinology; 2021

International Society of Endocrinology

Introduction: The relationship between the neuroendocrine and immune systems is essential for homeostaticmaintenance. We reported that physiological levels of triiodothyronine (T3) activate mice dendritic cells (DCs) throughthyroid hormone receptors (TRβ), inducing Th1 and Th17 proinflammatory responses and restraining tolerogenic signals.These results were exploited with success in an antitumor DC-based vaccination protocol in vivo. Besides, we describedthat in vitro, nano-sized extracellular vesicles (EVs) released by T3-stimulated DCs (T3-DCs) activate syngeneic DCs,contributing to paracrine DC communication. However, the immune modulatory role of T3-DCs derived EVs on T cells isstill unknown.Objectives: To evaluate the immune response induced by T3-DC released EVs in allogenic splenocytes.Methods: Bone marrow DCs obtained from C57BL/6 WT mice were stimulated (or not, control: C) with T3 (10nM) for18h. DC-secreted EVs (DCs-EVs) were isolated by differential ultracentrifugation of the supernatant (2,000g: 2K;10,000g: 10K; and 100,000g: 100K). Allogenic splenocytes were obtained from BALB/c mice and stimulated with DC-EVsfor 6 days. Proliferation, intracellular and secreted cytokine production were analyzed by flow cytometry and ELISAassays, respectively. Statistical analysis: Sidak´s multiple comparisons test. P