“Thyroid hormone transport and metabolism in mice dendritic cells: Expression of genes involved.”

GIGENA N; ALAMINO VA; MONTESINOS, MM; NAZAR, M; MASINI-REPISO AM; CREMASCHI, G; PELLIZAS CG

Florianópolis

Congreso; XV Congreso Latinoamericano de Tiroides (LATS).; 2013

Sociedad Latinoamericana de Tiroides (LATS)

TRANSPORT AND METABOLISM OF THYROID HORMONES ON DENDRITIC CELLS: EXPRESSION OF GENES INVOLVED. Nicolás Gigena; Alamino, VA; Montesinos, MM; Nazar, M; Masini-Repiso, AM; Cremaschi, G; Pellizas, CG. CIBICI-CONICET. UNC. ARGENTINA / BIOMED-CONICET. UCA. ARGENTINA Background: We demonstrated that murine DC expressing thyroid hormone receptor (HT) beta 1 (TRβ1) and that physiological levels of triiodothyronine (T3) DC inmature (DCi) stimulate maturation and enhance their capacity to stimulate T Li, leading to a Th1 cytokine response (Mascanfroni et al. FASEB J 2008, 22:1032). These effects were exerted by T3 through a AKT pathway and NF-kB-dependent (Mascanfroni et al. J Biol Chem 2010, 285: 9569). In turn we demonstrate that glucocorticoids are able to counteract the effects immunostimulatory of T3 on DC mature (DCm) and abolish its ability to polarize the adaptive immune response towards a Th1 profile (Montesinos et al. Steroids 2012, 77: 67). However, the molecular mechanisms involved in the transport and metabolism of HT in DC are unknown. Objetives: We further explored: 1) expression of the 3 HT transporters higher specificity Monocarboxylate Transporter 8 (MCT-8), MCT-10 and Organic Anion Transporting Polypeptides 1C1 (OATP1C1) in DC, 2 ) deiodinases expression of iodothyronines in DC: DIO 1, 2 and 3, at different stages of DC maturation, as well as to study the potential effect of T3 on the expression. Methods: DCi were pulsed with T3 (10 nM) or LPS (100 ng / ml) for 18 hours. The presence and levels of mRNA of the gene of interest mentioned above was evaluated through semi-quantitative RT-PCR. For its part, by qRT-PCR in real time (SYBR GREEN) determined the relative expression of mRNA for each gene. The identity of the gene of interest was confirmed by Nested RT-PCR and Genetic Sequencing. Results: 1) A. DC express only MCT-10 mRNA, not MCT-8 and OATP1C1, suggesting that the protein encoded function as HT transporter facilitating their entry into the cell; B. mRNA-MCT-10 expression in DCi is less than in DCm matured with stimuli such as LPS and T3. 2) A. assessed the expression of DIO 1, 2 and 3 in DC. This study revealed the presence of both mRNA Dio2 and DIO3 in DCi, not from DIO1; B. physiological levels of T3 regulate DIO3 expression at the transcriptional level by increasing the levels of its mRNA. Conclusions: This work demonstrated the presence of mRNA-MCT-10, suggesting that the protein coding function as HT transporter in DC. Meanwhile, we observed expression of mRNA-DIO2 and DIO3 in DCi but not DIO1, suggesting that DIO 2 and 3 take part in the activation and inactivation of HT inside the DC. In addition, we showed that active HT: T3, participates in the regulation of DIO 3 expression at the transcriptional level. Our findings provide evidence for the first time the presence of genes which are involved in the transport and metabolism of HT in DC.