IMMUNOMODULATORY ROLE OF SPHINGOSINE KINASE 1 (SK1) ON TRIIODOTHYRONINE (T3)-MATURED DENDRITIC CELLS (DC) AND THE DRIVEN ADAPTIVE RESPONSE

NEGRETTI BORGA, DANA MARÍA; BLANCO, ANTONELLA; PIRES TEIXERA, MARIANA; ALAMINO, VANINA ALEJANDRA; SOLER, MARÍA FLORENCIA; PUENTES, ELIDA NAHIR; DONADIO, ANA CAROLINA; CLARKE, CHRISTOPHER J. ; MONTESINOS, MARÍA DEL MAR; HANNUN, YUSUF A.; PELLIZAS, CLAUDIA GABRIELA

Mar del Plata

Congreso; Reunión Anual de Sociedades de Biociencia, LXVII Reunión Anual de la Sociedad Argentina de Investigación Clínica (SAIC).; 2022

Our group demonstrated that T3 generates adaptive proinflammatory and cytotoxic responses through DC activation, restraining regulatory signals. This protocol was successfully exploited in T3-stimulated DC (T3-DC)-based antitumor vaccines. T3 effects are mainly triggered by non-genomic mechanisms involving thyroid hormone receptor, Akt and NF-kB. Sphingosine-1-phosphate (S1P) is a bioactive sphingolipid produced by SK1 and 2. Although this pathway is involved in many proinflammatory conditions, little is known about its role in innate immune cells. We aim to evaluate the role of SK1 in T3-DC, and the driven adaptive immunity. Bone marrow DC were obtained from C57BL/6 WT or SK1-KO mice and stimulated (or not) with T3 (10nM). PF-543 (SK1 inhibitor, 100nM) was added, and 30 min later the T3 stimuli (PF-T3-DC). After 30 min, p-Akt/total Akt was analyzed by Western Blot. Allogenic splenocytes isolated from BALB/c mice were co-cultured with T3-DC or PF-T3-DC (exposed to T3 for 18h), for 3 days. Viability and proliferation were evaluated by FACS. Cytokines were measured by FACS and ELISA. Statistical analysis: Two-way ANOVA/Tukey test, and paired t test. p