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DC (dendritic cell) culture method, TLR (toll-like receptor) agonist and application of TLR agonist

A culture method and agonist technology, applied in the field of immune cells, can solve the problems of weak and slow antigen presentation ability

Inactive Publication Date: 2016-09-21
SHEN ZHEN ISTEM REGENERATIVE MEDICINE SCI TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, there are many expansion schemes for DC cells, such as the combination of cytokines IL-1β, IL-6, TNF-α, PGE-2, etc., or the combination of polyinosinic acid and bacterial lipopolysaccharide to promote DC cell maturation. DC maturation-promoting agents can only slowly promote DC cell maturation, and secrete pro-inflammatory cytokines at a low level, with weak antigen presentation ability

Method used

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  • DC (dendritic cell) culture method, TLR (toll-like receptor) agonist and application of TLR agonist
  • DC (dendritic cell) culture method, TLR (toll-like receptor) agonist and application of TLR agonist
  • DC (dendritic cell) culture method, TLR (toll-like receptor) agonist and application of TLR agonist

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Embodiment 1

[0052] 1. Preparation of TLR agonists

[0053] The TLR receptor involved in this embodiment is TLR7 / 8; the poly-T oligodeoxynucleotide (PolydT) used is purchased directly from Invitrogen; the imidazoquinoline derivative (CL075) used is purchased directly from Invitrogen , the chemical structural formula is as follows:

[0054]

[0055] (1) Preparation of CL075 aqueous solution: Dissolve 500μg CL075 powder in 1000μl water to make a stock solution with a concentration of 500μg / ml, then divide into 10 tubes and store at -20℃ for later use;

[0056] (2) Preparation of PolydT aqueous solution: Dissolve 100nmol PolydT liquid in 2ml of water to make a stock solution with a concentration of 50μmol / L, then divide into 10 tubes and store at -20°C for later use;

[0057] (3) Preparation of TLR agonist: Take 0.03ml of CL075 aqueous solution, 1.5ml of PolydT aqueous solution and 8.47ml of water and blend uniformly to obtain about 10ml of TLR agonist.

[0058] 2. Acquisition of immatur...

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Abstract

The invention discloses a DC (dendritic cell) culture method, a TLR (toll-like receptor) agonist and application of the TLR agonist. The TLR excitant comprises polymer T oligodeoxynucleotide, imidazoquinoline derivatives and solvents, wherein the concentration of the T oligodeoxynucleotide is 1 to 90 mu. mol / L; the concentration of the imidazoquinoline derivatives is 0.1 to 900 mu. g / ml; the structure formula is as shown in the description. The TLR agonist can promote the fast maturing of DCs; the DC antigen presentation capability is enhanced.

Description

technical field [0001] The invention relates to the technical field of immune cells, in particular to a method for culturing DC cells, a TLR agonist and applications thereof. Background technique [0002] TLR (Toll-like Receptors, Toll-like receptors) is an important class of protein molecules involved in nonspecific immunity, and it is also a bridge connecting nonspecific immunity and specific immunity. TLRs are single transmembrane non-catalytic proteins that recognize molecules with conserved structures derived from microorganisms. When microorganisms break through the body's physical barriers, such as skin, mucous membranes, etc., TLR can recognize them and activate the body to generate immune cell responses. Specifically, TLR is a type I transmembrane receptor, which consists of a cytoplasmic Toll-IL-1 receptor domain (Toll-IL-1 receptor domain, TIR domain), a transmembrane domain and an extracellular domain. The TIR domain provides a key point that can interact with ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0784
CPCC12N5/0639C12N2501/05C12N2501/056C12N2501/22C12N2501/2304
Inventor 曾宪卓唐熙
Owner SHEN ZHEN ISTEM REGENERATIVE MEDICINE SCI TECH CO LTD
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