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Method for recombining vaccinia virus by removing dominant epitope B8R and virus thereof

A vaccinia virus and dominant epitope technology, applied in the direction of virus/bacteriophage, biochemical equipment and methods, microorganisms, etc., can solve the problems that restrict the effectiveness of vaccines, reduce the effect of immune dominance, and increase the effect of immunogenicity

Inactive Publication Date: 2016-01-20
XIAN MEDICAL UNIV
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  • Abstract
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  • Claims
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Problems solved by technology

A prominent problem in current vaccine research and development is that the immunodominance of the carrier itself significantly inhibits the effective expression and immunogenicity of the foreign target antigen. The immunodominance effect of the carrier has become one of the main factors restricting the effectiveness of the vaccine.

Method used

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  • Method for recombining vaccinia virus by removing dominant epitope B8R and virus thereof
  • Method for recombining vaccinia virus by removing dominant epitope B8R and virus thereof
  • Method for recombining vaccinia virus by removing dominant epitope B8R and virus thereof

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Experimental program
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Effect test

Embodiment

[0049] The wild-type vaccinia virus WR strain in this embodiment is preserved in the Laboratory of Molecular Virus and Viral Immunology of Xi'an Medical College (this laboratory, the same below), and the recombinant vaccinia virus (VACV-OVA) with embedded OVA gene + ) was constructed by the applicant and deposited in this laboratory. Plasmid pSC11 was donated by Professor Bernard Moss of NIH in the United States, and cell lines CV-1 and BSC-1 were purchased from China Type Culture Collection. The mice used were female 6-week-old C57BL / 6 mice, weighing 18-22 g, purchased from the Animal Breeding Center of the Chinese Academy of Preventive Medicine, and raised in an SPF-grade mouse room with constant temperature and humidity.

[0050] Main reagents and materials:

[0051] DMEM, fetal calf serum, lipofectamine2000 transfection reagent were all purchased from Invitrogen Company, Taq enzyme, restriction endonuclease and T4DNA ligase were all purchased from NEB Company, virus genom...

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Abstract

The invention discloses a method for recombining a vaccinia virus by removing a dominant epitope B8R and a virus thereof. By means of the method, immunodominance of the vaccinia virus is reduced, and the safety of the vaccinia virus is improved. A plasmid pSB8R-OVA for replacing B8R with OVA is built and recombined with the wild type vaccinia virus (a WR strain) in a CV-1 cell, and a recombinant virus is obtained through screening and purifying. The influence on exogenous antigen immunogenicity by the dominant epitope B8R is eliminated, the immunodominance effect of the vaccinia virus can be effectively reduced, exogenous antigen immunogenicity is improved, and therefore the effectiveness of live vaccine with the vaccinia virus as a carrier is improved. Meanwhile, the virulence of the vaccinia virus serving as the vaccine carrier can be effectively reduced to improve safety of the vaccinia virus. By removing the dominant epitope of the vaccinia virus, a more effective carrier can be provided for vaccine and gene therapy.

Description

technical field [0001] The invention relates to the fields of biological genetic engineering and immunity, in particular to a method for recombining vaccinia virus by removing the dominant epitope B8R and its application. Background technique [0002] Immunodominance refers to the inconsistency of the body's presentation of antigenic epitopes. Pathogens can usually express hundreds of thousands of antigens, and the presentation of these epitopes by the body after infection with pathogens is not uniform. Some epitopes can induce a very strong immune response in the body, some can only cause a moderate immune response, and some Can only cause mild immune response or even no immune response. We refer to those epitopes that can induce a strong response as dominant epitopes (dominantepitope), and other epitopes as sub-dominantepitopes (sub-dominantepitope). Immunological dominance is usually described by a dominant hierarchy of the strength of the immune system's response to a ...

Claims

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Application Information

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IPC IPC(8): C12N7/01C12N7/02
Inventor 汪洋马茜许杰
Owner XIAN MEDICAL UNIV
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