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Subunit mixed vaccine of human enterovirus 71

A human enterovirus, hybrid vaccine technology for biomedical applications

Active Publication Date: 2012-08-29
INST OF LAB ANIMAL SCI CHINESE ACAD OF MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] However, when Japanese scholars immunized experimental monkeys with attenuated whole-virus vaccines, the animals still showed certain neurological symptoms, which raised doubts about the safety of whole-virus vaccines.

Method used

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  • Subunit mixed vaccine of human enterovirus 71
  • Subunit mixed vaccine of human enterovirus 71
  • Subunit mixed vaccine of human enterovirus 71

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Peptide Screening

[0029] step one

[0030] Segmentation and engineered expression of human enterovirus type 71

[0031] The name of the strain used for segmentation is Fuyang-0805a, and its storage number in NCBI is GU350629. The genome CDS region of the strain is 6582bp in total, encoding 2193 amino acids. The size of the split polypeptide is controlled at about 100 amino acids (aa), the lower limit is 22aa, and the upper limit is 156aa. There are 4 Overlap of ~15 aa, as shown in Table 1.

[0032] Table 1 Segmentation of human enterovirus type 71

[0033]

[0034] Add NdeI and SacI sites to both ends of the primers for amplifying the polypeptide coding region. The expression vector uses the modified pET28a(+) expression vector. After the vector is modified, the N-terminus of the expressed target protein is except for the 6×His tag protein tag. , does not contain redundant amino acid sequences.

[0035] The successfully constructed expression vector was trans...

Embodiment 2

[0044] Vaccine testing

[0045] The 15 weakly cross-reactive or non-cross-reactive polypeptides obtained above were combined into 18 combinations, as described in Table 2:

[0046] The combination of table 218 polypeptides

[0047]

[0048]

[0049] Then the above 18 kinds of combinations were carried out in equal amounts for subsequent effect verification:

[0050] The 18 combinations were used to immunize 6-week-old female mice respectively, and the immunization dose was 10 μg protein / mouse. The vaccine is mixed with complete Freund's adjuvant for the first immunization. Booster immunizations were performed three weeks after the initial immunization with incomplete Freund's adjuvant. One week after the booster immunization, female mice were mated.

[0051] The antibody titer, neutralizing titer and cross-reaction with human brain tissue were measured in the fourth, fifth and seventh weeks after the initial immunization respectively. It was found that all 18 candid...

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Abstract

The invention belongs to the biomedical field and provides a subunit mixed vaccine of the human enterovirus 71. The subunit mixed vaccine contains one or more of the polypeptides of the human enterovirus 71 polypeptide chain and necessary adjuvants, wherein the polypeptides are P1-69, P70-159, P140-249, P324-433, P444-565, P566-665, P746-876, P1197-1338, P1441-1526, P1549-1668, P1649-1731, P1732-1851, P1952-2071, P2072-2193 and P1843-1951. The vaccine has no potential toxic or side effect, has the protective function of anti-virus infection in vitro and in vivo and is a vaccine candidate withthe potential clinical application value.

Description

technical field [0001] The invention belongs to the field of biomedicine, in particular to a method for researching human enterovirus 71 vaccine. Background technique [0002] Human enterovirus type 71 (EV71) was isolated and identified for the first time in 1969 and is one of the pathogens of HFMD. The virus mainly infects children under the age of 5, mainly causing hand, foot and mouth disease and herpetic angina. In a few cases, complications include meningitis, pulmonary edema, hemorrhage, etc., and severe cases can lead to death. Human enterovirus 71 has caused many disease outbreaks worldwide. Since the 1980s, hand, foot and mouth disease caused by human enterovirus 71 has been raging in the Asia-Pacific region, causing serious infant deaths. [0003] The disease was discovered in Shanghai in my country in 1981, and has been reported in various provinces and cities since then. In March 2009, 54,713 cases of hand, foot and mouth disease were reported through the direc...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/125A61P31/14
Inventor 秦川张连峰刘江宁王卫
Owner INST OF LAB ANIMAL SCI CHINESE ACAD OF MEDICAL SCI
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