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HLA-A11 restricted GPC3-derived polypeptide and vaccine containing polypeptide

A technology of HLA-A11 and peptide vaccines, applied in the field of biopharmaceuticals, to achieve the effect of improving immune function

Inactive Publication Date: 2018-07-06
BEIJING YOUAN HOSPITAL CAPITAL MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0011] The HLA-A11 gene locus has a high expression rate in the Chinese population, however, its related restricted antigen vaccines are rarely reported, so the study of the HLA-A11 gene locus-restricted antigen vaccine has a high significance for the individualized immunotherapy of the Chinese population. The necessity of the research, and its research results have important and extensive application value

Method used

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  • HLA-A11 restricted GPC3-derived polypeptide and vaccine containing polypeptide
  • HLA-A11 restricted GPC3-derived polypeptide and vaccine containing polypeptide
  • HLA-A11 restricted GPC3-derived polypeptide and vaccine containing polypeptide

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Example 1 blood collection

[0048] The inventor of this application signed an informed consent form with the HLA-A11 liver cancer patient, and the patient was treated in the Tumor Interventional Therapy Center of Beijing You'an Hospital Affiliated to Capital Medical University. 10 ml of venous blood was drawn from the patient, and peripheral blood mononuclear cells were routinely isolated by Ficoll density gradient centrifugation according to a previously reported method.

Embodiment 2E

[0049] Example 2 ELISPOT method analysis of GPC3 specific killer T cell activity

[0050] The inventors of the present application isolated peripheral blood mononuclear cells from patients with HLA-A11 positive hepatocellular carcinoma and incubated them with GPC3 polypeptides, namely BJ4:443-451, BJ5:289-297, and BJ6:472-480, to detect whether there is GPC3 antigen specificity effector T cells.

[0051] The ELISPOT method was used to detect whether there were killer T cells that could specifically react with GPC3 and produce IFN-γ. Human IFN-γ ELISPOT kit (BD product) was used to detect whether IFN-γ was produced. If effector cells respond to GPC3-presenting cells and secrete IFN-γ, each IFN-γ molecule produces a brownish-red spot in the assay system.

[0052] First, the ELISPOT plate (BD) was coated with anti-human IFN-γ antibody for 18 hours, and blocked with 10% FCS RPMI1640 for 2 hours. Peripheral blood mononuclear cells (200 μl) were mixed with GPC3 peptide (10 μg) an...

Embodiment 3

[0053] Example 3 Detection of polypeptide immunogenicity

[0054] For immunization, polypeptide vaccines must have good immunogenicity and immunoreactivity, otherwise they cannot induce specific immune responses for immunoprophylaxis and immunotherapy. To this end, the inventors of the present application identified the polypeptide site, detected the immunoreactivity between the polypeptide and T cells of liver cancer patients, then inoculated the polypeptide into transgenic mice to detect the immunogenicity, and developed the HLA-A11-restricted GPC3-derived polypeptide vaccine in three steps .

[0055] In order to determine whether the polypeptide can induce GPC3 polypeptide-specific effector T cell responses when immunized as a vaccine, the inventors of the present application used human HLA-A11 transgenic mice to immunize with GPC3 polypeptide, and then immunized the polypeptide with the spleen of the immunized mice. The cells were co-cultured, and ELISPOT was used to dete...

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PUM

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Abstract

The invention discloses an HLA-A11 restricted GPC3-derived polypeptide and a vaccine containing the polypeptide, and belongs to the technical field of biological pharmacy. The HLA-A11 restricted GPC3-derived polypeptide is composed of an amino acid sequence of SEQ ID NO:1 or SEQ ID NO:2. The invention also discloses the vaccine containing the above polypeptide, an application of the above polypeptide in preparing an immune-inducing irritant of a GPC3-positive tumor or cancer, a preparation capable of inducing tumor-reactive T cells, a pharmaceutical composition used for treating and / or preventing the GPC3-positive tumor or cancer, an antibody, and an antigen presenting cell. The polypeptide provided by the invention can induce and produce a specific immune response, and can be used for immunoprophylaxis and immunotherapy.

Description

technical field [0001] The invention relates to an HLA-A11-restricted GPC3-derived polypeptide and a vaccine containing it, belonging to the technical field of biopharmaceuticals. Background technique [0002] Hepatocellular carcinoma (HCC) is one of the most common malignant tumors, and its incidence is particularly high in my country, and it is increasing year by year. About 110,000 people die of liver cancer every year in my country, accounting for 45% of the world's liver cancer deaths. The vast majority of HCC originate from liver cirrhosis, and hepatitis B is the main cause of liver cirrhosis nodules, and the malignant transformation of nodules eventually leads to HCC. Usually liver cancer does not show obvious symptoms until the advanced stage, when the tumor has metastasized or the liver function has deteriorated severely, and only about 20% of patients can be treated surgically. For this reason, new methods, including immunotherapy, have gradually become adjuvant ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/06A61K39/00A61P35/00C12N5/10C07K16/32C12N5/077
CPCA61K39/0011A61K2039/572C07K7/06C07K16/32C12N5/0639C12N5/0659C12N2510/00
Inventor 王延军孙斌陈德喜
Owner BEIJING YOUAN HOSPITAL CAPITAL MEDICAL UNIV
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