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CDK5 antigen epitope peptide and application thereof

An antigen epitope and antigen technology, applied in the field of molecular immunology, can solve the problem of insufficient immunogenicity of tumor-specific antigens

Inactive Publication Date: 2017-10-20
XIN HUA HOSPITAL AFFILIATED TO SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In view of the problems of insufficient immunogenicity of tumor-specific antigens in the prior art, the present invention provides a CDK5 epitope peptide and its application

Method used

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  • CDK5 antigen epitope peptide and application thereof
  • CDK5 antigen epitope peptide and application thereof
  • CDK5 antigen epitope peptide and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Example 1: Prediction and synthesis of HLA-A0201-restricted CDK5 protein CTL epitope peptide

[0044] (1) Determination of the amino acid sequence of CDK5 protein:

[0045] By searching the CDK5 protein sequence in the Genbank database, a CDK5 protein sequence with the Genbank accession number: CAG33322.1 was obtained, which consists of 292 amino acid residues, as shown in SEQ ID No.1.

[0046] (2) Online prediction of CDK5 protein epitope peptide:

[0047] Using the Epitope Peptide Prediction Database

[0048] a: https: / / www-bimas.cit.nih.gov / cgi-bin / molbio / ken_parker_comboform and

[0049] b: http: / / www.syfpeithi.de / bin / MHCServer.dll / EpitopePrediction.htm)

[0050] The service provided is to make a preliminary prediction of the HLA-A0201-restricted CDK5 protein epitope peptide, select several epitope peptides with the highest scores, and then use the supermotif and quantitative motif scheme to modify the initially predicted CTL epitope peptide , to further increas...

Embodiment 2

[0060] Example 2: T2 cell epitope peptide binding affinity and stability experiments

[0061] The affinity and stability of HLA-A0201 and peptides were determined with the help of the characteristics of T2 cells.

[0062] (1) T2 cell affinity experiment

[0063] Collect T2 cells, centrifuge and wash three times with sterile PBS at 4°C, add serum-free 1640 medium, 2×10 5 Cells per well were seeded in a 24-well cell culture plate, and an experimental group and a control group were set up. Three secondary wells were repeated in each group. T2 cells stimulated without peptide were used as blank controls, and corresponding experimental peptides (aa106 and aa164) and control peptide (positive control: HIV pol peptide; negative control: influenza virus matrix protein peptide FLU-matrix) (final concentration 50uM), while adding β2 microglobulin (final concentration 2.5ug / ml). Place cells at 37 °C, 5% CO 2 Incubate in the incubator for 18 hours, collect the cells again, wash three t...

Embodiment 3

[0070] Example 3: Detection of pentamer-positive cytotoxic T cells corresponding to peptide aa106

[0071] The corresponding pentamer of peptide aa106 is based on the principle of double recognition of T cell activation. The heavy chain α and β2 microglobulins of MHC class I molecules are assembled in vitro by bioengineering technology, and combined with the antigenic epitope peptide to form a TCR specific Sexually bound monomers, and then five monomers are assembled together to form a pentamer.

[0072] Two healthy donors and six patients with glioma who were diagnosed with non-remission were selected, and the CDK5 epitope peptide pentamer-positive CD8 in peripheral blood was detected. + number of cells. Collect 5ml of peripheral blood from healthy donors and patients with glioma diagnosed without remission, extract peripheral blood mononuclear cells with lymphocyte separation medium, count the cells and take 1×10 6 Dissolve in 100ul PBS, add 2ul peptide aa106 corresponding...

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PUM

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Abstract

The invention discloses antigen epitope peptide. An amino acid sequence contains an amino acid sequence shown in SEQ ID No.2 or SEQ ID No.3, or an amino acid sequence which is formed by adding, deleting or replacing one or multiple amino acids of the amino acid sequence shown in SEQ ID No.2 or SEQ ID No.3. The antigen epitope peptide can be used for producing antigen presenting cells, main tissue compatibility antigen compounds or cell toxic T lymphocyte inducing agents, and can be applied to prepare medicines for treating cancers, such as glioma.

Description

technical field [0001] The invention belongs to the field of molecular immunology and relates to antigenic epitope peptides, in particular to CDK5 epitope peptides recognized by HLA-A0201-restricted cytotoxic T lymphocytes (CTL) and applications thereof. Background technique [0002] Glioma is the most common malignant tumor of the nervous system, with a high recurrence rate and poor prognosis. The main reason is that the residual disease of the tumor is difficult to eliminate. Immunotherapy can further remove the minimal residual lesions after surgery, radiotherapy and chemotherapy, thereby reducing or avoiding the recurrence of glioma. However, finding tumor-associated antigens is the key to tumor immunotherapy. Researchers continue to screen new tumor-associated antigens to finally achieve the goal of individualized treatment for each patient. Studies have shown that cyclin-dependent kinase 5 (CDK5) is highly expressed in glioma and is associated with PD-L1 expression. ...

Claims

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Application Information

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IPC IPC(8): C12N9/12C12N15/54C12N5/0783A61K39/00A61P35/00
Inventor 孙媛陈书艳王飞
Owner XIN HUA HOSPITAL AFFILIATED TO SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE
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