Tumor antigenic polypeptide and application thereof as tumor vaccine

An antigen and vaccine technology, which can be used in anti-tumor drugs, medical preparations containing active ingredients, peptide sources, etc., and can solve problems such as strength differences

Inactive Publication Date: 2014-02-12
BEIJING ZHIFEI LVZHU BIOPHARM +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In fact, different HLA molecules can selectively bind peptides with different anchor positions and residues, so different HLA allele products may present different epitopes of the same antigen molecule, resulting in different individuals (with different MHC alleles) differ in strength of response to the same antigen

Method used

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  • Tumor antigenic polypeptide and application thereof as tumor vaccine
  • Tumor antigenic polypeptide and application thereof as tumor vaccine
  • Tumor antigenic polypeptide and application thereof as tumor vaccine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0073] The synthesis of embodiment 1 antigenic peptide

[0074] The synthetic polypeptide MTPGTQSPFFLLLLLLTVLTVVTGS (SEQ ID NO: 1), hereinafter referred to as antigenic peptide. The synthesized peptide was purified to 97%. Lyophilized antigenic peptides were dissolved in dimethyl sulfoxide (25 mM) and stored at -80°C.

[0075] The same method was used to synthesize polypeptides FLLLLLTVLT (SEQ ID NO: 2), LLLLLTVLTV (SEQ ID NO: 3), LLLLTVLTVV (SEQ ID NO: 4), FFLLLLLTVL (SEQ ID NO: 5), GTQSPFFLLL (SEQ ID NO: 6), TQSPFFLLLL (SEQ ID NO: 7), respectively referred to as antigenic peptide 1, antigenic peptide 2, antigenic peptide 3, antigenic peptide 4, antigenic peptide 5 and antigenic peptide 6.

Embodiment 2

[0076] Example 2 Preparation of dendritic cells (Dendritic Cells, DCs)

[0077] Mononuclear cells (PBMC) in peripheral blood include lymphocytes and monocytes, and their volume and shape specific gravity are about 1.075. Use the Ficoll-Hypaque separation medium with a specific gravity of 1.077 for density gradient centrifugation, so that cells with a certain specific gravity are distributed according to the corresponding density gradient, and various blood cells can be separated.

[0078]Obtain the white blood cell concentrate of healthy volunteers from the Hong Kong Red Cross Blood Center, dilute it with PBS to an appropriate multiple, and then add it to the surface of the Ficoll-Hypaque separation medium, centrifuge at 800g density gradient for 30min at 18-20°C, and after centrifugation is completed, the centrifuge tube The content is divided into three layers, the upper layer is plasma (containing platelets), the middle layer is separation fluid, and the bottom layer is red...

Embodiment 3

[0079] Preparation of embodiment 3 T cells (nylon cotton method)

[0080] T cells have short and few villi on the surface, while B cells have many and long villi. Due to the different smoothness of the cell surface, B cells are easy to attach to nylon cotton fibers at 37°C, while T cells do not have this ability. Using this property, T cells and B cells can be separated.

[0081] Preparation of nylon cotton column: After sterilizing 1g of nylon cotton / column, add RPMI1640 medium and place it at 37°C for 2 hours to allow RPMI1640 to flow out. Resuspend the above non-adherent cells in an appropriate volume of RPMI1640 so that the cell concentration is 0.5-1.0x10 8 / ml, then added to the prepared nylon cotton column, 2ml / column, incubated at 37°C for 1 hour, added RPMI1640 to the nylon cotton column, 10ml / column, eluted the cells not adsorbed on the nylon cotton, collected and eluted The solution is the T cell suspension. Centrifuge at 400g for 5min, resuspend the cell pellet ...

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Abstract

The invention provides a human mucin-1 tumor antigenic polypeptide with an amino acid sequence shown in SEQ ID NO: 1 or variants thereof, wherein the polypeptide can be combined with an HLA (Human Leukocyte Antigen) I and can be recognized by a cell CD8<+>T. The invention further provides nucleic acids coding the polypeptide. The invention further provides an antigen presenting cell capable of presenting the polypeptide on the surface of the cell and an immune effector cell capable of recognizing the polypeptide or antigen presenting cell. The invention further provides application of the polypeptide or variants thereof, the nucleic acids, the antigen presenting cell or the immune effector cell in the preparation of vaccines or pharmaceutical compositions for treating or preventing cancer. Tumor vaccines provided by the invention have a good treatment effect on relatively large crowds and are particularly applicable to the Asian, e.g. Chinese.

Description

technical field [0001] The present invention relates to the field of cancer immunity. In particular, the present invention relates to human mucin-1 tumor antigenic polypeptides and their use as tumor polypeptide vaccines. Background technique [0002] Among the diseases that endanger human health, tumor has become the main cause of human death. Globally, more than 10 million people are affected by cancer every year. According to the World Cancer Report of the World Health Organization, it is estimated that by 2020, the annual incidence will reach 15 million and 10 million people will die. The death rate of domestic cancer has risen to the first place in the city. This means that the situation facing tumor treatment is very severe. At present, the clinical treatment of cancer is still based on surgery, supplemented by chemotherapy and radiotherapy, but the specificity of the treatment is not enough, it often causes damage to normal tissues and organs, and surgical treatmen...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/47C12N15/12C12N5/0783C12N5/0784A61K48/00C12N5/10A61K39/00A61P35/00
CPCA61K39/00C07K14/4727C12N5/0636C12N5/0639
Inventor 谢雍杜琳
Owner BEIJING ZHIFEI LVZHU BIOPHARM
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