Vaccine

A technology of vaccines and vaccine products, applied in the field of vaccines, to achieve the effects of maintaining biological activity, enhancing specific CTL killing effect, and mild preparation process

Inactive Publication Date: 2009-12-16
INST OF PROCESS ENG CHINESE ACAD OF SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no effective treatment for rabies. Human rabies vaccine can stimulate the body to produce anti-rabies virus immunity, which is used to prevent rabies.

Method used

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Experimental program
Comparison scheme
Effect test

preparation example Construction

[0070] (1) For the preparation of splenocytes, see the ELSPOT procedure.

[0071] (2) Preparation of stimulating cells

[0072] ① Preparation of splenocytes from normal syngeneic mice, the method is the same as (1). Adjust the cell concentration to 5×10 with 1640 complete medium 6 cell / ml.

[0073] ② Add mitomycin C (MMC) to a final concentration of 25 μg / ml, at 37°C, 5% CO 2 Incubate for 2 hours.

[0074] ③ Add 30ml of serum-free 1640 medium, centrifuge at 400g for 10 minutes, and wash 3 times to remove residual mitomycin C.

[0075] ④Use 1640 complete medium containing 20U / ml rIL-2 to adjust the cell concentration to 5×10 6 cell / ml, add HBsAg MHC class I peptide S 28-39 , the final concentration was 20 μg / ml.

[0076] ⑤ at 5% CO 2 After incubating at 37°C for 4 hours in the incubator, the cell concentration was adjusted to 1×10 6 cell / ml.

[0077] (3) Preparation of effector cells

[0078] ① The splenocytes and stimulator cells of the immunized mice were mixed at ...

Embodiment 1

[0095]Add 10 mg of PLGA blank microspheres (average particle size of 1.5 μm, molecular weight of 10 kDa) to 1 ml of Hansen antigens with concentrations of 10 μg / ml, 40 μg / ml, 100 μg / ml, 200 μg / ml, and 400 μg / ml, The pH of the antigen solution is 5.5, and the concentration of NaCl in the solution is adjusted to be 0.9% w / v, and the oscillating adsorption is carried out at room temperature for 12 hours to obtain the vaccine product. ml, 400 μg / ml antigen was diluted to 40 μg / ml, each batch of microsphere antigen compound preparation and simple Hansen antigen were used to immunize mice with 4 μg antigen dosage. After 7 days, use the ELISPOT method to detect the secretion of IFN-γ in mouse spleen MNC, see figure 1 , the ordinate represents the number of cells secreting IFN-γ per 4×105 splenic MNCs stimulated by HBsAg MHC class I polypeptide S28-39, that is, the number of spot-forming cells (SFC); Effects on IFN-γ secretion of mice after 7 days, 1-10 μg / ml; 2-40 μg / ml; 3-100 μg / ml...

Embodiment 2

[0097] Three different particle sizes of PLA (molecular weight 15kDa) blank microspheres were used as vaccine adjuvants, with average particle sizes of 0.35 μm, 1.9 μm and 12.1 μm, respectively. figure 2 . Add 10 mg of the above three microspheres to 1 ml of Hansen antigen with a concentration of 45 μg / ml, adjust the pH of the antigen solution to 5.9, adjust the concentration of NaCl in the solution to 1.5% w / v, and oscillate for adsorption at 25°C 5h, each batch of microsphere antigen compound preparations were used to immunize mice with 4 μg of antigen, and 7 days later, the ELISPOT method was used to detect the secretion of IFN-γ in mouse spleen MNC, see image 3 . The ordinate represents the number of cells secreting IFN-γ per 4×105 splenic MNCs stimulated by HBsAg MHC class I polypeptide S28-39, that is, the number of spot-forming cells (SFC); The effect of IFN-γ secretion after 7 days, among which 1-0.35 μm microspheres; 2-1.9 μm microspheres; 3-12.1 μm microspheres; ...

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Abstract

The invention discloses a vaccine for prevention and treatment, wherein the antigen is in two forms with one absorbed on a biodegradable polymer particle and the other unabsorbed (free antigen). The average grain diameter of the particles is 0.1-20um; the antigen has specific structure and antiviral effect, preferably hepatitis B surface antigen and vaccines used for human being. The vaccine is prepared by the following methods: mixing polymer particles with antigen solution to obtain suspension under the absorption of the polymer particles and antigen solution, that is vaccine product; and adding cryoprotector in the obtained suspension-like vaccine to obtain lyophilized powder of the vaccine. The vaccine can induce immune response rapidly in the engine body, induces the engine body to generate high-level humor immune and cell immune, and eliminates or controls virus by secretoryantibody or cytokine. Besides, the vaccine has simple preparation process and mild preparation process.

Description

technical field [0001] The present invention relates to a vaccine, more specifically, to a vaccine with biodegradable polymer particles as an adjuvant. Background technique [0002] Hepatitis B (Hepatitis B) is a global infectious disease caused by hepatitis B virus (HBV). It has the characteristics of strong infectivity, high carrier rate and wide prevalence, and seriously endangers human health. According to the statistics of the World Health Organization (WHO), 2 billion people have been infected with hepatitis B virus in the world so far, of which about 350 million people are chronic hepatitis B infections. The total infection rate of hepatitis B virus in my country is 60%, and the carrier rate is about 10% (ie About 130 million), is a high prevalence area of ​​hepatitis B. There is no effective treatment drug for hepatitis B. Currently, interferon and lamivudine, which are commonly used in antiviral treatment, can quickly inhibit virus replication, but they are prone to...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/39A61K39/29A61K39/205A61P1/16A61P31/20A61P31/14
Inventor 马光辉卫强梁争论邱少辉苏志国李河民安康林小军范蓓田瑞魏炜
Owner INST OF PROCESS ENG CHINESE ACAD OF SCI
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