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Cell membrane lipids displaying gpi-ny-eso-1 fusion protein on the surface and application thereof

A technology of GPI-NY-ESO-1 and fusion protein, which is applied in the direction of non-active ingredient medical preparations, hybrid peptides, drug combinations, etc., can solve the problem that DCs cannot effectively present tumor cells, tumor cells escape the immune system, etc. question

Active Publication Date: 2016-06-29
BEIJING BIOHEALTHCARE BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Although DC has such a powerful immune surveillance function, due to some factors related to tumor cells, DC cannot effectively present tumor cells, so tumor cells escape the monitoring of the immune system

Method used

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  • Cell membrane lipids displaying gpi-ny-eso-1 fusion protein on the surface and application thereof
  • Cell membrane lipids displaying gpi-ny-eso-1 fusion protein on the surface and application thereof
  • Cell membrane lipids displaying gpi-ny-eso-1 fusion protein on the surface and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Example 1, Preparation of cell membrane lipids displaying GPI-NY-ESO-1 fusion protein on the surface

[0025] 1. Construction of recombinant plasmids and recombinant cells

[0026] 1. Insert the double-stranded DNA molecule (IgGFc-GPI fragment, 1374bp) shown in Sequence 3 of the sequence listing between the NheI and NotI restriction sites of the plasmid pCI-neo to obtain the recombinant plasmid pCI-pr-GPI.

[0027] 2. Insert the double-stranded DNA molecule shown in Sequence 2 of the sequence listing between the EcoRV and NheI restriction sites of the recombinant plasmid pCI-pr-GPI to obtain the recombinant plasmid pCI-pr-GPI / NY-ESO-1. According to the sequencing results, the structure of the recombinant plasmid pCI-pr-GPI / NY-ESO-1 is described as follows: a double strand shown in sequence 4 of the sequence table is inserted between the EcoRV and NotI restriction sites of the plasmid pCI-neo DNA molecule. The double-stranded DNA molecule shown in sequence 4 of the seq...

Embodiment 2

[0049] Example 2, Application of Cell Membrane Lipid Displaying GPI-NY-ESO-1 Fusion Protein on Surface and Cell Membrane Lipid Displaying GPI-NY-ESO-1 Fusion Protein on Surface with Oxidized Mannan Modification

[0050] Medium A-I: serum-free RPMI-1640 culture medium containing 30% (volume ratio) solution A-I.

[0051] Medium A-II: serum-free RPMI-1640 culture medium containing 30% (volume ratio) solution A-II.

[0052] Medium B-I: Serum-free RPMI-1640 culture medium containing 30% (volume ratio) Solution B-I.

[0053] Medium B-II: serum-free RPMI-1640 culture medium containing 30% (volume ratio) solution B-II.

[0054] Medium C (control medium): serum-free RPMI-1640 culture medium.

[0055] Medium D (control medium): Serum-free RPMI-1640 medium containing 50 μg / ml NY-ESO-1 polypeptide and 5 μl / ml Liposome 2000 (Invitrogen, product number 11668-019).

[0056] 1. Cell membrane lipids containing GPI-NY-ESO-1 fusion protein and DC cells promote human T lymphocytes to secrete c...

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Abstract

The invention discloses a cell membrane lipid with a surface displaying a GPI-NY-ESO-1 fusion protein and application thereof. Firstly, the cell membrane lipid with the surface displaying the GPI-NY-ESO-1 fusion protein is protected; the GPI-NY-ESO-1 fusion protein sequentially comprises the following components from upstream to downstream: an NY-ESO-1 polypeptide and a GPI fragment, wherein the NY-ESO-1 polypeptide is as shown in the section between amino acid residues at 2nd-26th sites from the N terminal of a sequence 5; the GPI fragment is as shown in the section between amino acid residues at 2nd-26th sites from the N terminal of the sequence 5. According to the invention, an oxidized mannan-modified cell membrane lipid with the surface displaying the GPI-NY-ESO-1 fusion protein is protected; the cell membrane lipid with the surface displaying the GPI-NY-ESO-1 fusion protein or the oxidized mannan modified-cell membrane lipid with the surface displaying the GPI-NY-ESO-1 fusion protein can outstandingly enhance the multiplication capacity of lymphocytes in cooperation with DC cell. the cell membrane lipid with a surface displaying a GPI-NY-ESO-1 fusion protein has great value in tumor therapy.

Description

technical field [0001] The invention relates to a cell membrane lipid displaying GPI-NY-ESO-1 fusion protein on the surface and application thereof. Background technique [0002] Tumor has become a major disease that seriously threatens human health in the 21st century. The tolerance of patients' immune system to tumor is an important reason for tumor occurrence and development. Dendritic cell (dendritic cell DC) is the most powerful antigen-presenting cell (APC) in the human body, which can regulate the body's immune response and initiate the specific killing effect of cytotoxic T cells on tumor cells. Because DC cells have immune stimulating ability, they are the only APCs found so far that can activate unsensitized naive T cells. DCs can not only activate T cells in an antigen-specific form, recognize and kill tumor cells, but also stimulate immune memory. Protection, which plays a protective role when the host is attacked by tumor cells again. DC is closely related to ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K19/00C12N15/85C07K1/107A61K39/385A61K47/48A61P35/00A61P37/04A61K35/15
Inventor 卢戌
Owner BEIJING BIOHEALTHCARE BIOTECH
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