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immune inducer

A technology of immune induction and sequence listing, applied in vaccines, blood cell cancer vaccines, DNA/RNA vaccination, etc.

Active Publication Date: 2022-07-22
TORAY IND INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no report that the MCEMP1 protein has immune-inducing activity against cancer cells, and thus this protein is useful for the treatment and prevention of cancer.

Method used

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Examples

Experimental program
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Embodiment 1

[0101] [Example 1] Obtaining novel cancer antigen protein by SEREX method

[0102] (1) Preparation of cDNA library

[0103] Total RNA was extracted from canine testis by the acid guanidium-phenol-chloroform method (Acid guanidium-Phenol-Chloroform method), and polyadenylic acid was purified using Oligotex-dT30 mRNA furification Kit (manufactured by Takara Shuzo Co., Ltd.) according to the protocol attached to the kit RNA.

[0104] A cDNA phage library was synthesized using the obtained mRNA (5 μg). For the preparation of the cDNA phage library, cDNA Synthesis Kit, Zap-cDNA Synthesis Kit, and ZAP-cDNA GigapackIII Gold Cloning Kit (manufactured by STRATAGENE) were used, and the library was prepared according to the protocol attached to the kit. The size of the prepared cDNA phage library is 1×10 6 pfu / ml.

[0105] (2) Screening cDNA library by serum

[0106] Immunoscreening was performed using the cDNA phage library prepared above. Specifically, the host Escherichia coli (...

Embodiment 2

[0114] [Example 2] Analysis of cancer antigenicity of MCEMP1 in vivo

[0115] (1) Production of recombinant vector expressing mouse MCEMP1 in vivo

[0116]Based on the nucleotide sequence of SEQ ID NO: 7, a recombinant vector expressing mouse MCEMP1 in vivo was prepared by the following method. PCR was performed using 1 μl of cDNA prepared from the mouse leukemia cell line EL4 (purchased from ATCC) whose expression was observed in Example 1, and two primers (described in SEQ ID NOs: 17 and 18) containing EcoRI and NotI restriction sequences. 0.4 μM, 0.2 mM dNTP, 1.25 U of PrimeSTAR HS polymerase (manufactured by Takara Shuzo Co., Ltd.) were added to make the total amount of each reagent and buffer solution 50 μl, and a thermal cycler (manufactured by BIO RAD) was used. The cycle of 55°C for 15 seconds and 72°C for 1 minute was repeated 30 times. In addition, the above-mentioned two kinds of primers are primers for amplifying the region encoding the full-length amino acid seq...

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Abstract

The present application relates to a novel immunity-inducing agent for the treatment and / or prevention of cancer, in particular to a polypeptide comprising at least one polypeptide selected from the group consisting of polypeptides derived from MCEMP1 and variants thereof and having immunity-inducing activity, or comprising a polypeptide encoding the same An immunity-inducing agent using a recombinant vector capable of expressing the polypeptide in vivo as an active ingredient, and an immunity-inducing method comprising the step of administering the immunity-inducing agent to a subject.

Description

technical field [0001] The present invention relates to novel immunity-inducing agents useful as cancer therapeutics and / or preventives. Background technique [0002] Cancer is the leading cause of death, and the current treatment is surgery, combined with radiotherapy and chemotherapy. Despite the development of new surgical methods and the discovery of new anticancer agents in recent years, the current situation is that, except for some cancers, the results of cancer treatment have not been greatly improved. In recent years, with advances in molecular biology and cancer immunology, cancer antigens recognized by cytotoxic T cells that respond to cancer, genes encoding cancer antigens, and the like have been identified, leading to expectations for antigen-specific immunotherapy improve. [0003] Mast Cell-Expressed Membrane Protein 1 (Mat Cell-Expressed Membrane Protein 1, MCEMP1) is a type 2 transmembrane protein, which has been reported to be specifically expressed in th...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/00A61K38/21A61K39/39A61K48/00A61P35/00A61P35/02A61P37/04C07K14/54C07K14/555C07K14/705C12N15/09A61K38/00A61K31/7088A61K35/76
CPCA61K38/00A61K38/21A61K31/7088C07K14/54C07K14/555C07K14/705A61K2039/804A61P35/00A61K39/001102A61K2039/5158A61K2039/53A61K2039/572A61K35/15A61K38/20A61K39/39A61K48/005C12N15/09A61K35/76
Inventor 藤田贵之冈野文义
Owner TORAY IND INC
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