DC (Dendritic Cell) vaccine as well as preparation method and application thereof

Abstract

The invention relates to a DC (Dendritic Cell) vaccine of a gastric cancer or a breast cancer as well as a preparation method and application thereof. A recombinant overlapped peptide fusion protein is constructed according to an amino acid sequence of a tumor antigen; the overlapped peptide fusion protein consists of polypeptides consisting of 15 amino acids in one series; two adjacent polypeptides have 11 amino acids which are completely overlapped and are the same; overlapped peptides are connected by the same enzyme cutting sites 1i-key; genes of the recombinant overlapped peptide fusion protein are cloned to a pET28 carrier through NdeI and BamH enzyme cutting sites; the recombinant overlapped peptide fusion protein is separated and purified after being expressed through escherichia coli; a PBMC (Peripheral Blood Mononuclear Cell) which is in-vitro separated from human blood, a GM-CSF (Granulocyte-Macrophage Colony-Stimulating Factor) and the recombinant overlapped peptide fusion protein are co-cultured; the recombinant overlapped peptide fusion protein generates a series of uncertain overlapped peptides after DC endocytosis and intracellular digestion, wherein the uncertain overlapped peptides include peptides of CTL (Cytotoxic Lymphocyte) phenotype and / or Th phenotype and are combined into MHC (Major Histocompatibility Complex) I and MHC II molecules of the DC, so that DC is matured to obtain the DC vaccine. Compared with a method for in-vitro synthesizing each peptide stimulating DC, the preparation method disclosed by the invention has quick stimulating DC benefits and low cost.

Description

[technical field] [0001] The invention relates to a DC vaccine and its preparation method and application, in particular to a gastric cancer or breast cancer DC vaccine and its preparation method and application. [Background technique] [0002] DC-based immunotherapy is one of the most valuable and promising approaches in tumor treatment today. Fusion of tumor cells and DCs, tumor antigen sensitization, and gene transfection have become effective approaches for tumor immunotherapy based on DCs. The modified DC can induce a specific anti-tumor immune response after reinfusion in vivo, and has achieved very valuable results in animal experiments and early clinical trials. [0003] 1) The treatment of hepatitis B virus may be rapidly developed due to the further understanding of the function and role of DC. Although some scholars have used specific DC to treat CHB patients, its safety and reliability need further evaluation, and the use of DC Basic research and animal experim...

AI Technical Summary

Problems solved by technology

However, for most DC vaccines for treating tumors, there are still some problems in the selection of antigens in theory: 1) Tumor lysates used as antigens are usually not easy to obtain a sufficient number of fragments, and may cause immune responses to self-antigens, Not suitable for clinical application

2) Synthetic antigenic peptides, due to the strict MHC restriction of the immune response, its clinical application is limited

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