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Kidney syndrome hemorrhagic fever mucosa immune vaccines and method of preparing the same

A technology for renal syndrome and mucosal immunity, applied in the field of vaccines, can solve the problems that mucosal immunity vaccines have not been reported yet

Inactive Publication Date: 2010-10-27
STATION OF VIRUS PREVENTION & CONTROL CHINA DISEASES PREVENTION & CONTROL CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Compared with traditional injection vaccines, mucosal vaccines have attracted more and more researchers' attention because of their advantages such as simplicity, economy, low pain, and avoidance of unclean vaccinations leading to disease transmission. Immunization vaccine has not been reported yet

Method used

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  • Kidney syndrome hemorrhagic fever mucosa immune vaccines and method of preparing the same
  • Kidney syndrome hemorrhagic fever mucosa immune vaccines and method of preparing the same
  • Kidney syndrome hemorrhagic fever mucosa immune vaccines and method of preparing the same

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Embodiment 1, the preparation of vaccine

[0028] 1. Expression and purification of Escherichia coli heat-labile enterotoxin B subunit

[0029] 1. Amplification of Escherichia coli heat-labile enterotoxin B subunit gene

[0030] 1) PCR amplification primers

[0031] Upstream primer: 5'-CAT CCATGG CTC CCC AGT CTA-3' (with Nco I site)

[0032] Downstream primer: 5'-TAA CTC GAG GTT TTC CAT ACT-3' (with Xho I site)

[0033] 2) PCR amplification conditions

[0034] The PCR reaction system is 100 μl, and the system composition is as follows:

[0035] 10× buffer: 10 μl

[0036] 4×dNTP (2.5mmol / L each): 5μl

[0037] Upstream primer (10pmol / L): 2.5μl

[0038] Downstream primer (10pmol / L): 2.5μl

[0039] Reaction template: Escherichia coli (E.coli) strain H 10407 Bacteria solution 2μl

[0040] High-fidelity DNA polymerase (5U / μl): 0.5μl

[0041] Double distilled water: 77.5 μl

[0042] The PCR reaction conditions are: 95°C for 40s, 52°C for 45s, 72°C for 45s, a t...

Embodiment 2

[0065] Embodiment 2, the inspection of vaccine

[0066] 1. Safety experiment of mucosal immunization vaccine for hemorrhagic fever with renal syndrome

[0067] Because there is E. coli heat-labile enterotoxin B subunit in the vaccine of the present invention, and E. coli heat-labile enterotoxin is one of the main toxins that cause diarrhea, although the B subunit is non-toxic in theory, but based on the present invention To be applied to the safety considerations of population vaccination, the toxicity evaluation was first carried out at the level of cells and experimental animals.

[0068] cellular level

[0069] Mammalian cell line Vero E6 was used, the positive control group was commercial cholera toxin 0.1 μg / mL, the sample groups were added with 10 μg / mL, 20 μg / mL, 50 μg / mL, 100 μg / mL LTB, and the positive control group started after 3 days of cell culture Symptoms such as cell detachment and shrinkage appeared, and the cells were obviously detached and died within 5 da...

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Abstract

The invention discloses a mucosa immune vaccine of hemorrhage fever syndrome and a preparation method thereof. The vaccine comprises inactivated virus and chitosan from hantaan virus which can cause hemorrhage fever syndrome in one or a plurality of types, wherein the chitosan exists in the form of micro-particles; the inactivated virus is covered inside the micro-particles of chitosan; furthermore, the vaccine can fruther comprise escherichia coli heat-labile enterotoxin B subunit with mucosa adjuvant activity; the result of the animal experiment shows that the vaccine of the invention can effectively excite the immune response of specific mucosa and the body fluid of the body anti-hantaan virus through nasal cavity and oral administration. The vaccine has statistical significance compared with the control group (p is less than 0.01) and has the advantages of safety, convenience, cost-effeciveness etc., compared with traditional hemorrhagic fever injected vaccine.

Description

technical field [0001] The invention belongs to the field of vaccines, in particular to a mucosal immune vaccine against hemorrhagic fever with renal syndrome and a preparation method thereof. Background technique [0002] Hantavirus is a segmented negative-strand RNA virus belonging to the genus Hantavirus in the family Bunyaviridae, and rodents are the main hosts of Hantavirus. According to serotype and genotype, Hantavirus can be divided into nearly 30 virus species (species). Hantavirus can cause two human diseases, namely, hemorrhagic fever with renal syndrome (Hemorrhagic Fever with Renal Syndrome, HFRS) and Hantavirus pulmonary syndrome (Hantavirus Pulmonary Syndrome, HPS). Hemorrhagic fever with renal syndrome is an acute viral infectious disease characterized by fever, bleeding tendency and kidney damage. According to international typing habits, Hantaan, Seoul, Dobrava, Puumala and other Hantaan viruses cause hemorrhagic fever with renal syndrome. Saaremaa type ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/12A61K47/36C12N7/06A61P31/14A61P13/12
Inventor 李德新曹守春张全福梁米芳李川刘琴芝
Owner STATION OF VIRUS PREVENTION & CONTROL CHINA DISEASES PREVENTION & CONTROL CENT
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