[35] To determine whether Notch activation was affected in Ts65Dn thymocytes, expression of selleck inhibitor the Notch target gene Hes-1 was measured in total thymus by quantitative PCR. Expression of Hes-1 was decreased 25% compared with euploid controls (Fig. 8a).

Similar changes were also observed in Lin− bone marrow cells (Fig. 8b). As an additional potential mechanism to down-regulate IL-7Rα levels, changes in miRNA expression levels were measured in Ts65Dn mice. Tissue samples from individuals with Down syndrome have increased expression of miRNAs encoded by the triplicated chromosome[36] and sequence analysis in the Ts65Dn mice indicated that the same miRNAs (miR-155, miR-125b, let-7c, miR-802 and miR-99a) are also encoded by the triplicated portion of MMU-16. Both miR-155 and miR-125b are known to be expressed in haematopoietic cells,[37] and analysis of the 3′-untranslated region of the IL-7Rα gene using TargetScan,[38] indicated that it contains consensus recognition sites for both miR-155 and miR-125b. Furthermore, B cells from transgenic mice over-expressing miR-155

had down-regulated IL-7Rα mRNA levels.[39] A significant increase in both miR-125b and miR-155 was observed in total thymocytes, Saracatinib mouse as well as in immature, DN thymocytes from Ts65Dn mice (Fig. 8c). Expression of miR-125b and miR-155 was also analysed in the bone marrow. The miR-155 expression was increased in both lineage-negative and total bone marrow samples in Ts65Dn mice in comparison to euploid mice, whereas

miR-125b expression was increased only in lineage-negative cells and not total bone Meloxicam marrow (Fig. 8d). Hence, decreased Notch activation and increases in miRNA may also contribute to the decreased levels of IL-7Rα expression in haematopoietic progenitors in the thymus and bone marrow. Although deficient immune responses and premature aging of the adaptive immune system has been reported for many years in DS, there is still controversy whether DS represents a model of immunosenescence or exhibits inherent immunodeficiency. Furthermore, underlying mechanisms that may affect lymphoid development and function have not been examined in depth. Older literature proposed changes in samples from individuals with DS, including altered thymic architecture and expression of adhesion molecules and inflammatory cytokines,[11, 40] whereas recent reports have focused upon defects in thymic gene expression[41] and thymic emigrants in human DS.[13, 14] Using the Ts65Dn mouse model to further define the changes in T-cell lineage development in DS, the data suggest that decreases in IL-7Rα expression in immature lymphoid cells lead to impaired thymic development. These data are consistent with previous observations in bone marrow progenitors,[12] and suggest a potential mechanism for immune alterations in DS that lead to a premature aging phenotype and senescence of peripheral lymphocytes. Similar to data in humans[12] and mice,[10] the Ts65Dn thymus was significantly smaller and hypocellular.