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Recombinant tubercle bacillus antigen poly epitope peptide and its constructing method and use

A technology of Mycobacterium tuberculosis and construction methods, applied in the field of molecular immunology, can solve problems such as death, and achieve the effect of promoting the resolution and promotion of multi-drug resistance problems

Inactive Publication Date: 2009-09-02
SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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  • Claims
  • Application Information

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Problems solved by technology

Intravenous injection of Mycobacterium tuberculosis into β2m-deficient mice can lead to rapid accumulation of bacteria in the liver, spleen, and lung tissues of mice, and soon lead to death

Method used

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  • Recombinant tubercle bacillus antigen poly epitope peptide and its constructing method and use
  • Recombinant tubercle bacillus antigen poly epitope peptide and its constructing method and use
  • Recombinant tubercle bacillus antigen poly epitope peptide and its constructing method and use

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Experimental program
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Embodiment 1

[0037] 1 Materials and methods

[0038] 1.1 Research object

[0039] 10 HLA-A*0201-positive healthy volunteers and 6 HLA-A*0201-negative healthy volunteers were all PPD+. 20ml of peripheral blood was collected and anticoagulated with heparin.

[0040] 1.2 Plasmids, strains and reagents

[0041] Plasmids pET22b and pGEX-4T-1, Escherichia coli DH5α and BL21 strains were purchased from Novagen

[0042] PCR kit: Promega

[0043] Plasmid mini-extraction kit, mini-gel recovery kit: Youjing Bioengineering Co., Ltd.

[0044] Restriction enzymes (BamH I, Nde I, Xho I): Takara

[0045] T4 DNA ligase: Roche

[0046] Agarose: Gibco

[0047] Tryptone, yeast extract: Oxoid

[0048] Isopropylthio-β-D-galactoside (IPTG): Shanghai Boguang Biotechnology Co., Ltd.

[0049] Nitrocellulose membrane: Amersham

[0050] Alkaline phosphatase (ACP)-labeled goat anti-human IgA: KPL company

[0051] Pre-stained protein marker: bio-lab company

[0052] Lymphoprep TM Purchased from Norway AXIS...

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Abstract

The present invention relates to the field of molecular immunology. Chinese patent application 200610027311.2 discloses 6 molecular mimetic peptides of antigenic epitopes that can stimulate a protective immune response against Mycobacterium tuberculosis in humans. On this basis, the present invention provides a recombinant Mycobacterium tuberculosis antigenic poly-epitope peptide, which contains SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5 and the fusion peptide of the amino acid sequence shown in SEQ ID NO:6. The present invention also provides construction methods and uses. The multi-epitope peptide has been verified by cell and animal experiments, and can stimulate an efficient and specific protective CTL immune response against tuberculosis infection. The invention contributes to the prevention and treatment of tuberculosis, especially the control of multidrug-resistant strains, and is also conducive to the study of molecular and cellular action mechanisms in tuberculosis, and promotes the solution of the multidrug-resistant problem that plagues the clinic.

Description

technical field [0001] The invention relates to the field of molecular immunology, in particular to a recombinant Mycobacterium tuberculosis antigen multi-epitope peptide capable of stimulating human body's protective immune response against Mycobacterium tuberculosis and its construction method and application. Background technique: [0002] Tuberculosis (TB) is one of the main killers threatening human health around the world. About 2 billion of the world's population has been infected with Mycobacterium tuberculosis (Mtb), and the vast majority of infected people have no symptoms and become latent infection (latentMtb.infection, LTBI), of which about 10% of latent infections can evolve into Tuberculosis, of which 16 million are active. Worldwide, about 8 million people are newly infected each year, and about 2 million people die from tuberculosis. In China, in the national infectious disease epidemic report released by the Ministry of Health in November 2005, the incide...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/195C12N15/62C12N15/31A61K39/04A61P31/06
Inventor 邓安梅仲人前娄加陶吴传勇陈孙孝周晔陈燕陈波蒋天舒谷明莉
Owner SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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