Attenuation bacteria carrier tuberculosis vaccine carrying tuberculosis, preparation method and application thereof

A tuberculosis vaccine, a technology for Mycobacterium tuberculosis, which is applied to the attenuated bacterial vector tuberculosis vaccine carrying tuberculosis genes and the fields of preparation and application, can solve problems such as deletion of ESAT6 gene, achieve stable expression, effective mucosal and systemic immune responses, and induce mucosal and the effect of the systemic immune response

Inactive Publication Date: 2009-05-13
WUHAN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] Bacillus Calmette-Guerin (BCG) is currently the only vaccine to prevent Mycobacterium tuberculosis infection, but it lacks the ESAT6 gene, which is a protective antigen

Method used

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  • Attenuation bacteria carrier tuberculosis vaccine carrying tuberculosis, preparation method and application thereof
  • Attenuation bacteria carrier tuberculosis vaccine carrying tuberculosis, preparation method and application thereof
  • Attenuation bacteria carrier tuberculosis vaccine carrying tuberculosis, preparation method and application thereof

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Embodiment 1

[0041] The preparation of attenuated Salmonella typhimurium vector vaccine Salmonella typhimurium SL7207 (pVAX-1-Ag85B) and Salmonella typhimurium SL7207 (pVAX-1-ESAT6-Ag85B) comprises the following steps:

[0042] A. Construction of eukaryotic expression plasmids pVAX-1-Ag85B and pVAX-1-ESAT6-Ag85B containing fusion genes of Mycobacterium tuberculosis H37Rv strain Ag85B and ESAT6-Ag85B (relevant literature published on volume 154 of "J Immunol."). Antigen 85B (Ag85B) and secreted 6-kDa protein (ESAT6) were obtained by PCR using the genome of Mycobacterium tuberculosis H37Rv strain as a template. Ag85B upstream and downstream primers are 5′-T GAATTC AAATGTTCTCCCGGCCG-3' and 5'-A AAGCTT TCAGCCGGCGCCTAA-3', (restriction sites are RamHI EcoRI), and then the obtained Ag85B and pVAX-1 were digested with BamHI and EcoRI respectively, and ligated to obtain pVAX-1-Ag85B eukaryotic expression plasmid; ESAT6 upstream and downstream primers 5′-GC AAGCTT ATGACAGAGCAGCAGTGGAA-3' and 5...

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Abstract

The invention discloses an attenuated bacteria vector tuberculosis vaccine carrying tuberculosis genes as well as a preparation method and application thereof. The preparation process includes the following steps: firstly, structuring a eukaryotic expression plasmid containing mycobacterium tuberculosis Ag85B and ESAT-6Agb5B genes; and secondly, using the electroporation method to convert the structure eukaryotic expression plasmid into attenuated Salmonella typhimurium which can stably express mycobacterium tuberculosis proteins. The combined utilization of Salmonella typhimurium SL7207 (pVAX-1-ESAT6-Ag85B) and BCG can achieve greater immune protective effect in resisting mycobacterium tuberculosis infection than by the BCG only; and the SIgA level of mucosa and serum produced by the Salmonella typhimurium SL7207 (pVAX-1-ESAT6-Ag85B) and BCG combined is higher than BCG and DNA vaccines. In addition, the Salmonella typhimurium SL7207 (pVAX-1-ESAT6-Ag85B) has greater immune protective ability than DNA vaccines or Salmonella typhimurium SL7207 (pVAX-1-Ag85B). The invention has the advantages of simple method, low cost, and convenient use, and can not only resist Salmonella typhosa infection, but also achieve immune protection against mycobacterium tuberculosis infection. Moreover, the stable expression of the mycobacterium tuberculosis proteins after being immunized can induce mucosal and systematic immune responses, and the invention has remarkable economic benefits.

Description

technical field [0001] The invention belongs to the field of molecular microbiology and infection immunity, relates to an attenuated Salmonella typhimurium vector vaccine carrying Mycobacterium tuberculosis genes ESAT6 and Ag85B, and relates to a preparation method of the vaccine. The recombinant attenuated Salmonella typhimurium vector vaccine can be used To resist the infection of Mycobacterium tuberculosis, it also has the application in immunity against Salmonella typhi infection. Background technique [0002] Tuberculosis (TB) is an infectious disease caused by Mycobacterium tuberculosis infection that seriously endangers human health. According to the statistics of the World Health Organization (WHO), nearly 1 / 3 of the people in the world are infected with Mycobacterium tuberculosis, 3 million people die from tuberculosis every year, and the number of new cases reaches 8 to 10 million every year. Bacillus Calmette-Guerin (BCG) is currently the only tuberculosis preven...

Claims

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Application Information

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IPC IPC(8): A61K39/04A61K39/112A61P31/06
CPCY02A50/30
Inventor 章晓联王其龙
Owner WUHAN UNIV
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