Mitogen-activated protein kinase pathway controls autoimmune regulator (AIRE) gene expression in granulo-monocyte colony stimulating factor (GM-CSF)-stimulated myelomonocytic leukemia OTC-4 cells

Autoimmune Regulator (AIRE) Gene Is Expressed in Human Activated CD4+ T‐Cells and Regulated by Mitogen‐Activated Protein Kinase Pathway

“…As shown in Fig. 2a, the AIRE gene was expressed in dendritic cells as previously reported (13) express the AIRE gene tempted us to examine the expression of the AIRE gene in antigen-and interleukin-2-stimulated T-cells, since the expression of the AIRE gene could be induced in GM-CSF-stimulated myelomonocytic OTC-4 cells (18).…”

“…Since the MAPK pathway controls AIRE gene expression in granulo-monocyte colony stimulating factor (GM-CSF)-stimulated OTC-4 cells and EBV-B cells (18), and both GM-CSF and IL-2 were known to mediate a similar cytokine signaling pathway (7,29), we next studied the role of the MAPK pathway in controlling the expression of the AIRE gene in IL-2T cells. As shown in Fig.…”

“…Instead, we used the CD4 ϩ Tcell isolation kit, which is an indirect magnetic labeling system for the isolation of untouched CD4 ϩ T-cells depleting non-CD4 ϩ cells with a cocktail of biotin-conjugated antibodies and anti-biotin microbeads. In addition, we screened 10 lots of fetal bovine serum to induce positive AIRE gene expression in the GM-CSFstimulated OTC-4 system (18,22) and a suitable lot of fetal bovine serum, lot no. C06177-500, MultiSer (ThermoTrace, Ltd., Melborne, Australia), was used throughout this study.…”

“…The myelomonocytic leukemia OTC-4 cell line was established from the peripheral blood of a patient with myelomonocytic leukemia (22). An EBV-transformed MDK-3B-cell line was established in our laboratory as described previously (18). Cells were grown in RPMI 1640 medium (Gibco) containing 10% fetal bovine serum (MultiSer, lot no.…”

Lymphotoxin Pathway and Aire Influences on Thymic Medullary Epithelial Cells Are Unconnected

“…Boehm et al (8) concluded that LT␤R signals are required for proper MEC differentiation and organization, while the second group, led by Fu (9,19,20), contended that signals through LT␤R regulate aire and PTA gene transcription and not thymus epithelium composition/structure. The latter view seems to have gained widespread acceptance (21)(22)(23)(24)(25)(26)(27)(28). In this study, we directly addressed these conflicting conclusions and further examined the role of the LT pathway in the thymus and its relationship to aire and PTA gene transcription, using a combination of histologic, flow cytometric, and gene expression-profiling approaches.…”