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Recombinant Mycobacterium smegmatis vaccine expressing HBHA-IL-12 fusion protein

A technology of Mycobacterium smegmatis and fusion protein, which is applied in the field of medicine and biology, can solve the problems that the treatment effect of MTB infection needs to be further improved, it is difficult to induce long-term effective immune response, and the residence time of subunit vaccine is short, so as to reduce the spleen load. Bacteria count, high safety, effect of clearing infection

Inactive Publication Date: 2013-02-06
FOURTH MILITARY MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

DNA vaccine is currently the most researched therapeutic vaccine, but its main problems are: the encoded antigen is relatively single, the immune response induced is very different from the natural infection process of MTB, multiple immunizations are required, and immune tolerance is easily induced and the production of autoantibodies; the residence time of subunit vaccines in the body is short, and it is difficult to trigger long-term effective immune responses; while live vector vaccines are superior to other genetically engineered vaccines in stimulating the body to produce protective immune responses, and are the focus of therapeutic vaccine research. an important direction
[0014] Although the above-mentioned protective antigens can induce effective cellular immune responses after immunization alone, their therapeutic effects on MTB infection still need to be further improved

Method used

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  • Recombinant Mycobacterium smegmatis vaccine expressing HBHA-IL-12 fusion protein
  • Recombinant Mycobacterium smegmatis vaccine expressing HBHA-IL-12 fusion protein
  • Recombinant Mycobacterium smegmatis vaccine expressing HBHA-IL-12 fusion protein

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Embodiment Construction

[0041] The present invention provides a recombinant Mycobacterium smegmatis vaccine expressing HBHA-IL-12 fusion protein. On the basis of constructing exogenous expression vector pSMT3-HBHA-IL-12, it is transfected into Clado smegmatis Bacillus prepared recombinant M.S, and HBHA-IL-12-rM.S was obtained through resistance screening; and the expression of the target gene and the immune effect of HBHA-IL-12-rM.S were verified, and the results showed that the vaccine could be used for MTB Infection treatment. The present invention will be further described in detail below in conjunction with the accompanying drawings, which are explanations rather than limitations of the present invention.

[0042] 1. Cloning of HBHA gene

[0043] The HBHA gene was cloned by PCR using MTB H37Rv DNA as a template, and the amplification primers were designed as follows:

[0044] Upstream primer P1: gc ggatcc at ggctgaaaac tcgaacattg 30;

[0045] Downstream primer P2: at gtcgac ct tctgggtgac c...

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Abstract

The invention discloses a recombinant Mycobacterium smegmatis vaccine expressing an HBHA-IL-12 fusion protein. The vaccine takes Mycobacterium smegmatis as a host cell, and an exogenous expression vector for transfecting the host cell contains HBHA-IL-12 gene segments. The recombinant Mycobacterium smegmatis vaccine can continuously express the bioactive HBHA-IL-12 fusion protein, and the induced IFN-gamma can effectively remove the infected MTB in a mouse. The vaccine combines the advantages of a target antigen and the live vector, and can be used for treating MTB infection.

Description

technical field [0001] The invention belongs to the technical field of medicine and biology, and relates to a recombinant mycobacterium smegmatis (rM.S) vaccine, in particular to a recombinant mycobacterium smegmatis vaccine expressing HBHA-IL-12 fusion protein. Background technique [0002] 1. A therapeutic vaccine for TB is urgently needed [0003] At present, about 1 / 3 of the world's population is in a state of persistent infection with Mycobacterium tuberculosis (M.tuberculosis, MTB), and 5% to 10% of infected people may eventually develop into active tuberculosis (TB) and become MTB. important source of infection. Chemotherapy is the basic treatment for tuberculosis. Although some active TB patients can be cured by this method, due to the long treatment time and serious side effects, the patient's compliance is poor, and it is easy to lead to the occurrence of drug resistance and persistent infection. Therefore, there is an urgent need to develop drugs or therapeutic ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/04A61K48/00A61P31/06C12N15/31C12N15/63
Inventor 师长宏张海徐志凯赵勇
Owner FOURTH MILITARY MEDICAL UNIVERSITY
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