Anti-Thyroid Peroxidase monoclonal antibody, clone FQS6480

Exosomes have been intensively studied in autoimmune diseases, and circulating exosomes and microvesicles have also been explored in autoimmune thyroiditis (AITD). However, the role of thyroid cell-derived exosomes in immune responses is unclear. We showed that IFN-gamma-treated Nthy-ori 3-1 cell-derived exosomes (IFN-gamma-Exo) harbored TPO, HSP60 and MHC-II and activated dendritic cells (DCs) in vitro. Compared with Exo-targeted DCs (DCExo), IFN-gamma-Exo-targeted DCs (DCIFN-gamma-Exo) promoted the expression and release of proinflammatory cytokines, such as IFN-gamma, IL-17A and IL-22, from CD4(+) T lymphocytes and inhibited the expression and release of antiinflammatory cytokines, such as IL-4, IL-10 and TGF-beta 1; however, IFN-gamma-Exo did not have this effect compared with Nthy-ori 3-1 cell-derived exosomes (Exo). DCIFN-gamma-Exo stimulates the expression and release of cytokines from CD4(+) T lymphocytes more efficiently than FN-gamma-Exo. Thus, DC(IFN-gamma-Exo )may effectively induce CD4(+) T lymphocyte-mediated immune responses and play a role in the occurrence and development of AITD.

Platelet function tests utilizing agonists or patient serum are generally performed to assess platelet activation ex vivo. However, inter-individual differences in platelet reactivity and donor requirements make it difficult to standardize these tests. Here, we established a megakaryoblastic cell line for the conventional assessment of platelet activation. We first compared intracellular signaling pathways using CD32 crosslinking in several megakaryoblastic cell lines, including CMK, UT-7/TPO, and MEG-01 cells. We confirmed that CD32 was abundantly expressed on the cell surface, and that intracellular calcium mobilization and tyrosine phosphorylation occurred after CD32 crosslinking. We next employed GCaMP6s, a highly sensitive calcium indicator, to facilitate the detection of calcium mobilization by transducing CMK and MEG-01 cells with a plasmid harboring GCaMP6s under the control of the human elongation factor-1 alpha promoter. Cells that stably expressed GCaMP6s emitted enhanced green fluorescent protein fluorescence in response to intracellular calcium mobilization following agonist stimulation in the absence of pretreatment. In summary, we have established megakaryoblastic cell lines that mimic platelets by mobilizing intracellular calcium in response to several agonists. These cell lines can potentially be utilized in high-throughput screening assays for the discovery of new antiplatelet drugs or diagnosis of disorders caused by platelet-activating substances.