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Polypeptide and nanometer particles thereof for promoting dendritic cells to take in antigen peptides and applications thereof

A technology of dendritic cells and nanoparticles, applied in the fields of biological sciences and drug carriers, can solve the problems of narrow non-toxic concentration range of cytotoxicity, shorten the active period of contrast agent, accelerate the biodegradation of contrast agent, etc., and achieve the goal of inhibiting tumor growth The effects of enhanced capacity, excellent physical and chemical properties, and convenience for large-scale production

Active Publication Date: 2014-07-09
HUAZHONG UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the current DC labeling methods have certain limitations: (1) The labeling efficiency is low: most of them last for more than 6 hours; (2) Most of the existing transfection reagents are highly positively charged and have potential cell Toxicity and a narrow non-toxic concentration range; (3) Probes stay in lysosomes for a long time to accelerate the biodegradation of contrast agents; (4) Due to the low ability of mature DC to phagocytize antigens, most of the labeling work Both are carried out before the immaturity of DC, but the maturation-promoting incubation time after labeling greatly shortens the effective activity period of the contrast agent

Method used

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  • Polypeptide and nanometer particles thereof for promoting dendritic cells to take in antigen peptides and applications thereof
  • Polypeptide and nanometer particles thereof for promoting dendritic cells to take in antigen peptides and applications thereof
  • Polypeptide and nanometer particles thereof for promoting dendritic cells to take in antigen peptides and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] A polypeptide that promotes the uptake of antigenic peptides by dendritic cells. The peptide is composed of α-helical polypeptides, linking sequences and antigenic peptides in series in the form of covalent bonds. The amino acid sequence of the α-helical polypeptide is: FAEKFKEAVKDYFAKFWD, the amino acid sequence of the connecting sequence is GSG, and the amino acid sequence of the antigenic peptide is: KVPRNQDWL. The amino acid sequence of the polypeptide that promotes the uptake of antigen peptides by dendritic cells is described in SEQ ID NO.1 in the sequence listing.

[0037] Preparation of nanoparticles carrying melittin, the steps are:

[0038] (1) will contain 3μmol DMPC

[0039] (1,2-dimyristoyl-sn-glycero-3-phosphocholine), 0.1 μmol cholesteryl oleate (Cholesteryl oleate, CO for short), 0.3 μmol DiR-BOA, 9.6 nmol PEGylated phospholipid in chloroform solution were fully mixed in a glass test tube uniform;

[0040] (2) dry the chloroform in the test tube with ...

Embodiment 2

[0050] For the comparison of the antigen peptide delivery ability of immature DC (imDC) and mature DC (mDC) with the nanoparticles that promote the uptake of antigen peptide by dendritic cells prepared in Example 1, see Image 6 . Image 6 Nanoparticles that promote the uptake of antigenic peptides by dendritic cells can be efficiently fluorescently labeled on both immature DCs and mature DCs (mDCs), and are more efficient on mature DCs. Figure 7 A comparison of the optical labeling efficiency of mDCs between nanoparticles that promote the uptake of antigenic peptides by dendritic cells and those that were not loaded with antigenic peptides was shown. DC targeting ability. Optical imaging can be performed after optically labeling mDCs with nanoparticles loaded with antigenic peptides, Figure 9 Confocal optical microscopy imaging showed that the nanoparticles loaded with antigenic peptides could transport the antigenic peptides into the cells instead of the cell membrane, w...

Embodiment 3

[0052] For the comparison of the antigen peptide delivery ability of the nanoparticles that promote the uptake of antigen peptide by dendritic cells and the free antigen peptide to mDC prepared in Example 1, see Figure 8 . Figure 8 It is shown that whether at high concentration or low concentration, mature DCs are significantly more effective in promoting the uptake of antigenic peptide nanoparticles by dendritic cells than free antigenic peptides, proving that nanoparticles that promote dendritic cell uptake of antigenic peptides can effectively promote mDC The uptake of antigenic peptides provides the basis for its application in DC vaccine therapy for tumors.

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PUM

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Abstract

The invention discloses a polypeptide and nanometer particles thereof for carrying antigen peptides, preparation methods and applications thereof, and belongs to the field of biological science and drug carriers. The polypeptide is formed by series connection of an alpha-helix polypeptide, a connecting sequence and an antigen peptide in a covalent bond manner, wherein the amino acid sequence of the alpha-helix polypeptide is FAEKFKEAVKDYFAKFWD, the amino acid sequence of the connecting sequence is GSG, and the connecting sequence of the antigen peptide is KVPRNQDWL. The polypeptide and the nanometer particles prepared from the polypeptides are capable of effectively improving the tumor antigen peptide uptake efficiency of the dendritic cells and promoting the prevention and treatment effect of tumors after the DC (dendritic cell) takes in the antigen peptides.

Description

technical field [0001] The invention belongs to the fields of biological science and drug carrier, and in particular relates to a polypeptide for promoting dendritic cells to take up antigenic peptides, a nanoparticle for promoting dendritic cells to take up antigenic peptides and applications thereof Background technique [0002] Antigenic peptides are short peptides prepared by chemical synthesis techniques according to the amino acid sequence of a known or predicted epitope in the pathogen antigen gene. The antigenic peptide can be directly used as a vaccine or can be used as a carrier by dendritic cells (DC). When used directly as a tumor vaccine, tumor antigen polypeptides are first taken up by antigen-presenting cells (APCs), mainly dendritic cells (DCs) and macrophages), and processed by APCs to form major histocompatibility molecular complexes (MHC), which is then recognized by the T cell receptor (TCR), generating an antigen-specific signal. When dendritic cell DC...

Claims

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Application Information

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IPC IPC(8): C07K19/00A61K39/00A61K47/48A61K49/00A61P35/00
Inventor 骆清铭张智红金红林钱源
Owner HUAZHONG UNIV OF SCI & TECH
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