Application of NY-ESO-1 for enhancing CA9 immunogenic molecular adjuvant

A NY-ESO-1, immunogenic technology, applied in the field of biomedicine, can solve problems such as affecting the natural history of tumors

Inactive Publication Date: 2017-05-31
宁波美丽人生医药生物科技发展有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Furthermore, the reduction of spontaneous NY-ESO-1 antigen in melanoma suggests t

Method used

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  • Application of NY-ESO-1 for enhancing CA9 immunogenic molecular adjuvant
  • Application of NY-ESO-1 for enhancing CA9 immunogenic molecular adjuvant
  • Application of NY-ESO-1 for enhancing CA9 immunogenic molecular adjuvant

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0065] NY-ESO-1 protein can form multimeric structures even in loading buffer in the presence of regular concentrations of β-mercaptoethanol.

[0066] Experiments have found that NY-ESO-1 proteins obtained from bacteria and mammalian cells can form multimeric structures even in loading buffers with conventional concentrations of β-mercaptoethanol ( figure 1 A, B).

Embodiment 2

[0068] The polymerization reaction of NY-ESO-1 is mediated by its intermolecular disulfide bonds.

[0069] Cysteine ​​in amino acid residues 75, 76, 78, 152 and 165 were replaced with serine. Three types of NY-ESO-1 with amino acid substitutions were: ESOcs1 with cysteine-serine substitutions at positions 75, 76, and 78, ESOcs2 with substitutions at positions 152 and 165, and ESOcs3 with mutations at all five positions. Purify these proteins and use Western blot to compare the oligomerization state between them ( figure 1 B). The results showed that the wild-type protein clearly showed the presence of dimers, tetramers and even multimers of 130 kDa. ESOcs1 and ESOcs2 showed mainly monomer and dimer structures, while the result of ESOcs3 showed mainly monomer. This experiment shows that the polymerization of NY-ESO-1 is due to the disulfide bonds between its molecules.

Embodiment 3

[0071] NY-ESO-1 binds to human and mouse immature dendritic cells in vitro and is related to its own polymeric structure.

[0072] Purify these proteins and use Western blot to compare the oligomerization state between them ( figure 1 B). The results showed that the wild-type protein clearly showed the presence of dimers, tetramers and even multimers of 130 kDa. ESOcs1 and ESOcs2 showed mainly monomer and dimer structures, while the result of ESOcs3 showed mainly monomer. This experiment shows that the polymerization of NY-ESO-1 is due to the disulfide bonds between its molecules. NY-ESO-1 present in cancer cells was also analyzed by Western blot for its protein aggregation in loading buffer containing conventional concentrations of β-mercaptoethanol, and it was found that NY-ESO-1 mainly manifested as trimer and tetramer Dimeric and monomeric forms are rarely found (mutation 1C). Experiments show that NY-ESO-1 binds to human and mouse immature dendritic cells in vitro is ...

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Abstract

The invention relates to the field of biological medicine, and especially relates to an application of NY-ESO-1 for enhancing a CA9 immunogenic molecular adjuvant. According to the invention, a poly structure is formed in NY-ESO-1 protein even in a beta-mercaptoethanol loading buffer having normal concentration; a polymerization of NY-ESO-1 is mediated by a disulfide bond between molecules; combination of NY-ESO-1 and unmaturated dendrite cell of human and mice in vitro is related with a self polymerization structure; in vitro combination of NY-ESO-1 and DC cell in marrow is influenced by TLR4; combination of NY-ESO-1 and unmaturated dendrite cell of human and mice can be effected through calprotectin; polymer oligomerization reduces binding capacity of TLR4 and NY-ESO-1; the polymerization structure of NY-ESO-1 and TLR4 in a host participate in an immuno-sphere antibody reaction; and increase of Art v1(wtArt) and CA9 gene immunogenicity depends on NY-ESO-1 gene fusion expression. The NY-ESO-1 has effect for enhancing the CA9 immunogenic molecular adjuvant.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to the role of NY-ESO-1 as a molecular adjuvant for enhancing the immunogenicity of CA9. Background technique [0002] It has been postulated that innate immune system receptors may be involved in the initial autoimmune response to tumor-specific antigens (TAAs). Necrotic tumor cells release endogenous factors or damage-associated pattern molecules such as heat shock proteins, high mobility group box B1 (HMGB-1), and uric acid. These damage-patterning molecules alert the innate immune system via CD91, receptor for advanced glycation end products, TLR2 / TLR4, and interleukin-1 (IL-1) receptor, respectively. TAA itself is generally regarded as a bystander of the above-mentioned "danger signal", acting as an adjuvant in the initial stage of the anti-tumor immune response. According to this example, the spontaneous immune response is caused by new peptides produced by mutations in 11,000 gen...

Claims

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Application Information

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IPC IPC(8): A61K39/39
Inventor 田晓丽
Owner 宁波美丽人生医药生物科技发展有限公司
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