Antituberculosis vaccine as well as preparation method and application thereof

A tuberculosis and vaccine technology, applied in the field of tuberculosis vaccine, can solve the problems of being unsuitable for immunocompromised patients and ineffective in adults, and achieve the effects of effective and durable immunity, fast onset of action, and good immunogenicity

Active Publication Date: 2018-11-06
WEST VAC BIOPHARMA CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The technical problem to be solved by the present invention is: the existing vaccine is only effective for children, but not effective for adults; or it is not suitable for immunocompromised patients, etc.

Method used

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  • Antituberculosis vaccine as well as preparation method and application thereof
  • Antituberculosis vaccine as well as preparation method and application thereof
  • Antituberculosis vaccine as well as preparation method and application thereof

Examples

Experimental program
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Effect test

preparation example Construction

[0041] The invention provides a kind of preparation method of anti-tuberculosis vaccine, comprises the following steps:

[0042] a. Obtain mycobacterial bacterial strains, inoculate and culture them to the logarithmic growth phase, add resuspension medium, homogenize, sieve, and obtain single-celled mycobacterial cells;

[0043] b. The anti-tuberculosis vaccine is obtained by irradiating mycobacterium single-cell thalline with low dose of radiation, intermittently, uniformly and circularly.

[0044] Wherein, in the preparation method of the above-mentioned anti-tuberculosis vaccine, the resuspension medium described in step a is phosphate buffered saline (PBST) with Tween added. Further, the resuspension medium is PBS containing 0.05%-0.1% Tween 80.

[0045] Wherein, in the preparation method of the above-mentioned anti-tuberculosis vaccine, the concentration of mycobacterium single-cell thalline described in step a is 10 6 / ml~10 8 / ml.

[0046] Wherein, in the preparatio...

Embodiment 1

[0073] Example 1 Screening of the irradiation mode of anti-tuberculosis vaccine

[0074] C57BL / 6 mice were selected and divided into model group (PBS), XBCGa group (uniform irradiation; the dose rate was constant at 19Gy / min, 20min each time, 9 times in total, and the total amount of irradiation was 3420Gy), XBCGd group (decreasing radiation irradiation; the dose rate decreased from 35Gy / min to 19Gy / min and 12Gy / min in turn, each dose rate was irradiated 3 times, 20min each time, a total of 9 times, the total amount of irradiation was 3960Gy), XBCGi group (increased irradiation ; The dose rate increased from 12Gy / min to 19Gy / min and 35Gy / min, each dose rate was irradiated 3 times, 20min each time, a total of 9 times, the total amount of irradiation was 3960Gy), BCG group (BCG, Commercially available), 10 rats in each group, subcutaneously immunized once, injection volume 0.1mL (about 10 6 CFU / mL). After 4 weeks of immunization, the virus was challenged intravenously with fre...

Embodiment 2

[0078] Example 2 Observation of Immune Protection by Irradiation of Anti-TB Vaccine of the Present Invention

[0079] C57BL / 6 mice were selected and divided into model group (PBS), XBCGa group (uniform irradiation), XBCGd group (decreasing irradiation), XBCGi group (increasing irradiation), BCG group, 10 mice in each group, subcutaneously immunized for 1 times, injection volume 0.1mL about 10 5 CFU. After 4 weeks of immunization, the virus was challenged intravenously with freshly cultured BCG, and the challenge dose was 10 6 CFU, the spleen tissue homogenate was collected and counted with 7H10 at the 4th week after challenge.

[0080] The result is as figure 2 Shown:

[0081] The experiment showed that compared with the model group, the CFU of the XBCGa group and the BCG group decreased significantly at the 4th week, while the XBCGd group and the XBCGi group had no significant change compared with the model group. The results suggested that the anti-tuberculosis vaccine...

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Abstract

The invention belongs to the field of tuberculosis vaccines, and particularly relates to an antituberculosis vaccine as well as a preparation method and application thereof. Aiming at the problems that an existing vaccine has a poor effect on adults or is unsuitable for hypoimmunity patients and the like, the invention provides the preparation method of the antituberculosis vaccine. The preparation method comprises the following steps: firstly, obtaining a mycobacterium monocell thallus; secondly, interruptedly and circularly irradiating the monocell thallus by adopting rays at low dose to obtain a mycobacterium vaccine, wherein the rays are X-rays, gamma rays or rays generated by an isotope radioactive source Co60; the radiation dose rate lies in that radiation is performed at the rate of10 to 20Gy / min for 8 to 10 times at an interval of 5 to 10 minutes each radiation of 20 minutes. According to the antituberculosis vaccine disclosed by the invention, all antigen characteristics of the thallus are completely retained, stronger specific immune response can be excited more quickly, and effective durable immunity is obtained. The prepared vaccine has the advantages of low toxicity,quick onset and higher safety, and can be used for preventing and treating tituberculosis of immunocompromised patients.

Description

technical field [0001] The invention belongs to the field of tuberculosis vaccines, and in particular relates to an anti-tuberculosis vaccine and its preparation method and application. Background technique [0002] Epidemic status of tuberculosis: Tuberculosis (TB) is a serious infectious disease worldwide. According to the 2016 WHO report, there were 10.4 million new TB cases worldwide, and children accounted for 10%. Among them, there were 480,000 cases of multidrug-resistant tuberculosis (MDR), and another 100,000 cases of rifampicin-resistant tuberculosis. my country is a country with a high burden of TB. WHO predicts that the characteristics of tuberculosis infection and incidence in countries with a high burden of tuberculosis from 2016 to 2020 will be multidrug-resistant tuberculosis (MDR-TB) and AIDS combined with tuberculosis (TB / HIV). [0003] Current status of tuberculosis prevention and control: Bacillus Calmette-Guérin vaccine (BCG) is currently the only way ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/04A61K39/39A61P31/06A61P11/00A61P25/00A61P15/00A61P19/08A61P1/00
CPCA61K39/04A61K39/39A61P1/00A61P11/00A61P15/00A61P19/08A61P25/00A61P31/06A61K2039/521A61K2039/5154A61K31/4409A61K31/7036A61K31/496A61K2039/572A61K9/0019A61K9/0053A61K9/0043C12N13/00A61K2300/00A61K9/007A61K2039/54A61K2039/542A61K2039/543A61K2039/544C12N1/20
Inventor 王震玲魏于全
Owner WEST VAC BIOPHARMA CO LTD
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