Plasmacytoid dendritic cells having immune tolerance, and method for producing same
A technology of dendritic cells and immune tolerance, applied in the direction of blood/immune system cells, biochemical equipment and methods, animal cells, etc., can solve problems such as unclear systemic and standardized differentiation methods of immune tolerance pDC , to achieve the effect of stabilizing supply and inhibiting expression
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[0045] The present invention relates to a method for preparing immune-tolerant plasmacytoid dendritic cells comprising the step of treating immature dendritic cells with a Toll-like receptor agonist.
[0046] In this specification, the term "dendritic cell (DC)" refers to a cell that responds to various antigens together with MHC (major histocompatibility complex) class I complex or MHC class II complex by absorbing the antigen into the cell. The sample is presented to professional antigen presenting cells of T cells. In addition, dendritic cells include immunogenic (immunogenic) and immune tolerance (tolerogenic) antigen-presenting cells, and can be divided into immature dendritic cells (immature dendritic cells; "imDC"), semi-mature dendritic cells, etc. Semimature dendritic cells ("smDC") and mature dendritic cells ("mDC").
[0047] In this specification, the term "immature dendritic cells" are found in the precocious stage of dendritic cells, do not express CD14 which is ...
Embodiment 1
[0088] Example 1: Regulating effect of Toll-like receptor agonists on differentiation of plasmacytoid dendritic cells
[0089]To confirm whether treatment of Toll-like receptor agonists during differentiation of immature dendritic cells induces differentiation into immune-tolerant plasmacytoid dendritic cells, according to figure 1 The design diagram shown is as follows for experiments.
[0090] Specifically, the bone marrow of the thigh of C57BL / 6 mice was sampled using a bone marrow sampling syringe. After washing the sampled bone marrow with phosphate buffer saline (PBS), ammonium chloride was used to remove red blood cells. Separated cells (3×10 6 cell / well) inoculated into a 6-well plate, add 10% FBS (Fetal bovine serum, calf serum), 2mM L-glutamine, 100U / ml penicillin / streptomycin, 50μM mercaptoethanol , 0.1 mM non-essential amino acids, 1 mM sodium pyruvate, and 250 ng / ml of FLT3L in 1 ml of RPMI 1640 and start medium differentiation. On the third day from the ini...
Embodiment 2
[0092] Example 2: Confirmation of Stimulation and Cytokine Secretion Profiles of Isolated Plasmacytoid Dendritic Cells
[0093] In order to confirm the cytokine secretion morphology of the plasmacytoid dendritic cells isolated in Example 1 above, the isolated cells (5×10 5 cells / ml) were inoculated into a 48-well plate, and treated with ODN1826 (1 μg / ml) to generate stimulation. After 24 hours, the obtained supernatant was separated, and the cytokine secretion pattern was confirmed by enzyme-linked immunosorbent assay (ELISA, Enzyme Linked Immunosorbentassay), and the results were shown in Figure 4 . However, for the treatment effect of the Toll-like receptor agonist, the case where the Toll-like receptor agonist was not treated during the differentiation of immature dendritic cells in the above-mentioned Example 1 was represented as a comparative example (pDC).
[0094] Such as Figure 4 As shown, in general, type I interferon (IFN-α and IFN-β) and representative inflam...
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