Immunomodulatory role of sphingosine kinase 1 (SK1) on triiodothyronine (T3)-matured dendritic cells (DC) and the driven adaptive response.

NEGRETTI BORGA DANA; BLANCO ANTONELLA; MARIANA TEXEIRA; ALAMINO VANINA; FLORENCIA SOLER, MARÍA; PUENTES ELIDA; DONADIO ANA CAROLINA; CLARKE CHRISTOPHER; MARIA DEL MAR MONTESINOS; YUSUF HANNUN; PELLIZAS CLAUDIA GABRIELA

Congreso; Reunión Annual de Sociedades de Biociencias SAIC, SAI&FAIC, SAFIS.; 2022

SAIC, SAI&FAIC, SAFIS.

Our group demonstrated that T3 generates adaptive proinflammatoryand cytotoxic responses through DC activation, restraining regulatorysignals. This protocol was successfully exploited in T3-stimulatedDC (T3-DC)-based antitumor vaccines. T3 effects are mainlytriggered by non-genomic mechanisms involving thyroid hormonereceptor, Akt and NF-kB. Sphingosine-1-phosphate (S1P) is a bioactivesphingolipid produced by SK1 and 2. Although this pathwayis involved in many proinflammatory conditions, little is known aboutits role in innate immune cells. We aim to evaluate the role of SK1in T3-DC, and the driven adaptive immunity. Bone marrow DC wereobtained from C57BL/6 WT or SK1-KO mice and stimulated (or not)with T3 (10nM). PF-543 (SK1 inhibitor, 100nM) was added, and 30min later the T3 stimuli (PF-T3-DC). After 30 min, p-Akt/total Aktwas analyzed by Western Blot. Allogenic splenocytes isolated fromBALB/c mice were co-cultured with T3-DC or PF-T3-DC (exposedto T3 for 18h), for 3 days. Viability and proliferation were evaluatedby FACS. Cytokines were measured by FACS and ELISA. Statisticalanalysis: Two-way ANOVA/Tukey test, and paired t test. p