The Autoimmune Regulator (
Aire) protein coordinates the negative selection of developing thymocytes by inducing the expression of hundreds of tissue-specific
antigens within the thymic medulla, which is also a primary site of the expression of the immune checkpoint
HLA-G molecule. Considering the immunomodulatory properties of Aire and
HLA-G, and considering that the role of the constitutive thymus expression of
HLA-G has not been elucidated, we studied the effect of AIRE
cDNA transfection on
HLA-G expression in 4D6 thymic cells and in the
HLA-G-positive JEG-3
choriocarcinoma cells. Aire promoted the transactivation of
HLA-G gene by increasing the overall transcription, inducing the transcription of at least G1 and G2/G4
isoforms, and incrementing the occurrence and distribution of intracellular
HLA-G protein solely in 4D6 thymic cells.
Luciferase-based assays and
chromatin immunoprecipitation experiments performed in 4D6 cells revealed that Aire targeted at least two regions within the 5'-untranslated regulatory region (5'-URR) extending 1·4 kb from the first ATG
initiation codon. The interaction occurs independently of three putative Aire-binding sites. These results indicate that the Aire-induced upregulation of
HLA-G in thymic cells is likely to act through the interaction of Aire with specific
HLA-G 5'-URR
DNA-binding factors. Such a multimeric transcriptional complex might operate in the thymus during the process of promiscuous gene expression.