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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

Pending Publication Date: 2019-10-15
QURATIS INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In particular, although many studies demonstrating the important role of pDCs in various autoimmune or infectious diseases have been conducted, systematic and standardized differentiation methods for the preparation of immune-tolerant pDCs are still unclear

Method used

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  • Plasmacytoid dendritic cells having immune tolerance, and method for producing same
  • Plasmacytoid dendritic cells having immune tolerance, and method for producing same
  • Plasmacytoid dendritic cells having immune tolerance, and method for producing same

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Embodiment approach

[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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Abstract

The present invention relates to a method for producing plasmacytoid dendritic cells having immune tolerance, plasmacytoid dendritic cells having immune tolerance produced by the method, and a cell therapeutic agent comprising the cells. The present invention can induce a high yield of plasmacytoid dendritic cells having immune tolerance from immature dendritic cells through a simple and easy process, thereby enabling a stable supply of the plasmacytoid dendritic cells having immune tolerance in large quantities.

Description

technical field [0001] The present invention relates to a method for preparing immune-tolerant plasmacytoid dendritic cells, immune-tolerant plasmacytoid dendritic cells prepared according to the method, and a cell therapy agent comprising the same. Background technique [0002] In the immune system, immune response and immune tolerance coordinate and maintain balance. In the case of an excessive immune reaction, rheumatoid arthritis, autoimmune disease and sepsis of type 1 diabetes, and allergic diseases can occur, which have a great impact on human survival and quality of life. [0003] Dendritic cells were previously thought to play an important role only in inducing immune responses, but recently their research has become more active as it has become known that they also contribute significantly to immune tolerance responses. [0004] Dendritic cells are cells that play a central role in the regulation of innate immunity and acquired immunity due to acquired causal sele...

Claims

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Application Information

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IPC IPC(8): C12N5/0784A61K35/15
CPCA61K35/15A61K38/18C12N5/064C12N5/0639C12N2506/11C12N2502/1114C12N2501/999C12N2501/26C12N2501/998
Inventor 慎城宰金愚植金洪敏
Owner QURATIS INC
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