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Chimpanzee adenovirus vector based Ebola virus vaccine

An Ebola virus, chimpanzee technology, applied in the field of immunology

Inactive Publication Date: 2018-03-06
INST PASTEUR OF SHANGHAI CHINESE ACADEMY OF SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The replicative virus vector vaccine represented by rVSV-ZEBOV jointly developed by Merck Sharp&DohmeCorp, NewLink Genetics and Public Health Agency of Canada has entered the clinical phase III, and a single immunization has good immunogenicity, but there are safety problems; by The replication-deficient adenovirus vector vaccine represented by ChAd3.EBOZ / ChAd3.EBO jointly developed by Glaxo Smith Kline and NIH has entered clinical phase II, and there is no safety problem, but a second immunization is required to increase its immunogenicity ; Ad26-ZEBOV, Ad5-EBOV and other vaccines based on replication-defective adenovirus vectors have also entered clinical research; Recombinant DNA vaccines represented by Etc. and others have entered clinical phase I, but electroporation technology and secondary immunization are required for immunization

Method used

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  • Chimpanzee adenovirus vector based Ebola virus vaccine
  • Chimpanzee adenovirus vector based Ebola virus vaccine
  • Chimpanzee adenovirus vector based Ebola virus vaccine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0094] Example 1. Construction of recombinant chimpanzee adenovirus plasmid DNA pAdC68-EBOVgp and pAdC7-EBOVgp and virus titer detection

[0095] Such as figure 1 As shown, the entire expression element of EBOVgp containing CMV promoter and poly A tail was cloned into AdC68 and AdC7 vectors respectively to obtain recombinant chimpanzee adenovirus plasmid DNA, namely pAdC68-EBOVgp ( figure 1 A) and pAdC7-EBOVgp ( figure 1 B).

[0096] The full-length nucleotide sequence of pAdC68-EBOVgp is shown in SEQ ID NO:2; the full-length nucleotide sequence of pAdC7-EBOVgp is shown in SEQ ID NO:3.

[0097] First the inventors will EBOVgp fragment ( figure 2 The fragment obtained in A) was cloned into the pShuttle vector, and then the entire expression element ( figure 2 The fragments obtained in B) were cloned into AdC68 and AdC7 vectors respectively, and identified by various restriction enzymes ( figure 2 C), indicating that the inventors successfully constructed recombinant ade...

Embodiment 2

[0099] Embodiment 2, the expression detection of the EBOVgp protein of recombinant chimpanzee adenovirus

[0100] The inventors used 10 8 , 10 9 , 10 10 vp recombinant chimpanzee adenovirus AdC68-EBOVgp and AdC7-EBOVgp, and 10 9 , 10 10 AdC68-empty and AdC7-empty empty control of vp were used to infect HEK293 cells respectively. After 24 hours of infection, the cells were lysed on ice with protein lysate containing protease inhibitors, and the protein supernatant was obtained by centrifugation, and then the expression of EBOVgp protein was detected by Western assay. . Such as image 3 As shown, the HEK293 cells infected with different titers of AdC68-empty and AdC7-empty blank control could not detect the expression of EBOVgp; while the infection of 10 9 , 10 10 The recombinant chimpanzee adenovirus AdC68-EBOVgp of vp and HEK293 cells of AdC7-EBOVgp can detect the obvious expression of EBOVgp protein. The codon-optimized EBOVgp sequence can be expressed efficiently whe...

Embodiment 3

[0102] Example 3, Detection of specific antibody against Ebola GP protein after mice were immunized with recombinant chimpanzee adenovirus

[0103] The inventor according to Figure 4 The indicated mouse immunization schedules were used to immunize different treatment groups of mice. The result is as Figure 5 As shown, higher titer can be detected in the serum of mice in AdC68-EBOVgp group (primary immunization of AdC68-EBOVgp, second immunization of PBS) and AdC7-EBOVgp group (primary immunization of AdC7-EBOVgp, second immunization of PBS) after 2 weeks of primary immunization. Anti-Ebola GP protein-specific antibody ( Figure 5 A); AdC68-EBOVgp group, AdC7-EBOVgp group and AdC7-EBOVgp+AdC68-EBOVgp group (primary immunization AdC7-EBOVgp, second immunization AdC68-EBOVgp) mouse serum can also A higher titer of specific antibody against Ebola GP protein was detected ( Figure 5 B); AdC68-EBOVgp group, AdC7-EBOVgp group and AdC7-EBOVgp+AdC68-EBOVgp group can still detect ...

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Abstract

The invention relates to a chimpanzee adenovirus vector based Ebola virus vaccine. The chimpanzee adenovirus vector based Ebola virus vaccine is characterized in that replication-defective chimpanzeeadenovirus vector is utilized to build a novel Ebola virus vaccine. The vaccine vector is applicable to preparation of a virus vaccine with efficient expression and high immunogenicity. The Ebola virus vaccine is simple to prepare, low in cost, high in safe, and free from additives.

Description

technical field [0001] The invention belongs to the field of immunology, and more particularly, the invention relates to an Ebola virus vaccine based on a chimpanzee adenovirus vector. Background technique [0002] Ebola virus disease (EVD), also known as Ebola hemorrhagic fever (Ebola hemorrhagic fever, EHF), is a highly fatal infectious disease caused by Ebola virus (EBOV). In 1976, Ebola hemorrhagic fever first broke out simultaneously in the Yambuku area of ​​the Democratic Republic of the Congo (formerly Zaire) and the Nzara area of ​​Sudan. Since then, the Ebola virus has caused multiple epidemics around the world. From 2014 to 2016, several countries in West Africa, including Guinea, Libya, Sierra Leone and Nigeria, experienced the largest outbreak of Ebola hemorrhagic fever with the largest number of infections and deaths in history. According to the statistics of the World Health Organization (WHO), as of March 27, 2016, there were a total of 28,646 cases of infect...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/12A61P31/14C12N15/861
CPCA61K39/12C12N15/86C12N2710/10043C12N2760/14134
Inventor 周东明万里明宋宇峰杨茜
Owner INST PASTEUR OF SHANGHAI CHINESE ACADEMY OF SCI
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