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

CN101816783BInactive Publication Date: 2013-02-06FOURTH MILITARY MEDICAL UNIVERSITY
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Patents(China)
Current Assignee / Owner
Publication Date
2013-02-06
Estimated Expiration
Not applicable · inactive patent

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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.
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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 ...

Examples

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...