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DC-based HCV epitope vaccine and preparation method thereof

An epitope vaccine and virus technology, applied in the fields of bioengineering and biomedicine, can solve the problems of weak immunogenicity, unstable properties, weak specificity, etc., and achieve good regulation, strong immunogenicity and specificity, and good properties. stable effect

Inactive Publication Date: 2015-11-11
SHEN ZHEN ISTEM REGENERATIVE MEDICINE SCI TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The technical problem to be solved by the present invention is to provide a stable character, strong immunogenicity and specificity based on the DC HCV Viral Epitope Vaccine

Method used

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  • DC-based HCV epitope vaccine and preparation method thereof
  • DC-based HCV epitope vaccine and preparation method thereof
  • DC-based HCV epitope vaccine and preparation method thereof

Examples

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Effect test

preparation example Construction

[0025] A kind of HCV virus epitope vaccine based on DC provided by the present invention, its preparation method comprises the following steps:

[0026] 1) CD34 + and / or CD14 + Monocyte induction culture into immature DC;

[0027] 2) When the immature DC concentration is less than or equal to 5×10 5 When cells / ml, add HCV virus antigen polypeptide to co-culture with immature DC;

[0028] 3) Add tumor necrosis factor-α and interleukin or lipopolysaccharide and continue to culture for 7-13 days, until the DC matures and the DC is loaded with HCV virus antigen polypeptide.

[0029] In step 1), CD34 provided by the present invention + and / or CD14 + The method of monocyte induction and culture into immature DC is: CD34 + and / or CD14 + Monocytes were inoculated into medium containing granule-macrophages and interleukin-4 and incubated at 37°C, CO 2 After 6-9 days of induction at a concentration of 5%, immature DCs can be obtained. The phenotype and function of immature DCs a...

Embodiment 1

[0038] The preparation method of the DC-based HCV virus epitope vaccine comprises the following steps:

[0039] 1) After taking human peripheral blood, CD14 was screened by magnetic beads + cells, the CD14 + Cells were inoculated into medium containing 800 U / mL granulo-macrophage factor and 1000 U / mL interleukin-4 and incubated at 37°C, CO 2 Under the condition of 5% concentration, induce for 6-9 days to obtain immature DC;

[0040] 2) Wait until the DC concentration reaches 5×10 5 1 / ml, co-cultivate 1 ml of immature DCs and HCV virus antigen polypeptides composed of HCVNS3 and HCVNS5 polypeptide chains;

[0041] 3) Add 5 ng / ml tumor necrosis factor-α and 8 ng / ml interleukin-1 to continue culturing for 7 days until DCs are mature and loaded with HCV virus antigen polypeptides.

Embodiment 2

[0043] The difference from Example 1 provided by the present invention lies in that in this example, the concentration of tumor necrosis factor-α is 10 ng / ml, and the concentration of interleukin-1 is 5 ng / ml.

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Abstract

The invention relates to a DC-based HCV epitope vaccine and a preparation method thereof. The preparation method comprises 1, carrying out CD34<+> and / or CD14<+> cell induced culture to obtain immature DC, 2, adding linear polypeptide comprising HCVNS3 and HCVNS5 polypeptide chains into the immature DC and carrying out co-culture, and 3, adding a tumor necrosis factor-alpha and interleukin or lipopolysaccharide into the culture and carrying out culture for 7-13 days until the DC is mature. The DC-based HCV epitope vaccine has stable properties and strong immunogenicity and can well adjust a HCV virus-resistant body immunization mechanism.

Description

technical field [0001] The invention relates to the fields of bioengineering and biomedicine, in particular to a method for inducing transformation of umbilical cord mesenchymal stem cells into islet cells. Background technique [0002] Hepatitis C virus (HCV) is one of the main pathogens that cause chronic liver disease. There are about 170 million to 200 million HCV infected people in the world. The HCV infection rate in my country is about 3.2%. It is estimated that the infected people in the whole country are more than 40 million. After HCV infection, about 50%-85% turn into chronic hepatitis, 20%-30% of which develop into liver cirrhosis, and some turn into hepatocellular carcinoma (HCC), which seriously endangers human health and has become a global and severe public health problem. Hygiene issue. So far, there is still a lack of ideal antiviral treatment measures, and interferon treatment is not satisfactory. Due to the high mutation rate of HCV virus, the research ...

Claims

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Application Information

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IPC IPC(8): A61K39/29A61P31/14C12N5/0784
Inventor 曾宪卓鲁菲
Owner SHEN ZHEN ISTEM REGENERATIVE MEDICINE SCI TECH CO LTD
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