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Compound plague vaccine and preparation method thereof

A plague vaccine and vaccine technology, which is applied in the field of plague vaccines, can solve the problems of short protection period, high side reaction rate, and difficulty in vaccine transportation, and achieve the effects of reducing side effects, improving safety, and improving immune effect

Inactive Publication Date: 2012-07-25
NAT INST FOR THE CONTROL OF PHARMA & BIOLOGICAL PROD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Its disadvantages are: the vaccine is expensive; the protection period is short, and it needs to be injected every year; the side effects are high, the incidence of local reactions is 11% to 24%, and the incidence of systemic reactions is 4% to 6%.
Aluminum salt is the only adjuvant widely used in human beings. It has the advantages of low cost and large-scale production, but its use for many years has shown that it has many shortcomings. The effect is not satisfactory enough; it can cause local irritation and inflammation; moreover, the vaccine with aluminum salt as adjuvant cannot be freeze-dried and must be stored in cold storage, which causes difficulties in the transportation of the vaccine; aluminum salt is easy to accumulate in the body, especially in the brain, and has May cause neurological disorders
[0005] Both the traditional dead plague vaccine and the live vaccine have many shortcomings, such as high side reaction rate and unsatisfactory effect

Method used

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  • Compound plague vaccine and preparation method thereof
  • Compound plague vaccine and preparation method thereof
  • Compound plague vaccine and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Embodiment 1: the preparation of the plague vaccine that the bacterial cell lysate of attenuated plague EV strain and recombinant V antigen and F1 antigen form

[0040] 1. Preparation of lysate of plague attenuated EV strain

[0041] 1.1. Bacterial culture: Dissolve the attenuated plague EV strain (from Lanzhou Institute of Biological Products, strain number 52006) preserved at low temperature, inoculate it on Hou's agar medium, and cultivate it at 28°C for 48 hours to form the first generation. The first generation was replanted on Hou's agar medium, and cultured at 28°C for 48 hours was the second generation, and the second generation was replanted on Hou's agar medium, and cultured at 28°C for 72 hours was the third generation.

[0042] 1.2. Bacteria collection: wash down the thalli on the surface of Thicket's agar medium with a freeze-dried protection solution (prepared according to the preferred method mentioned above).

[0043] 1.3. Preparation of the homogenate ...

Embodiment 2

[0051] Embodiment 2 Safety test

[0052] In order to verify the safety of the above bacterial cell homogenate+V+F1 vaccine, the experiment was divided into four groups, each with 10 male BALB / c mice, respectively:

[0053] 1) Negative control group: normal saline 0.1ml / piece

[0054] 2) Cell homogenate group: cell homogenate 56.4μg / 0.1ml / piece

[0055] 3) Thale homogenate + V + F1 vaccine group: Thale homogenate + F1, V 10μg / 0.1ml each

[0056] 4) Live attenuated plague EV vaccine: 1×10 7 / 0.1ml / piece

[0057] Subcutaneous immunization was adopted on the back and observed for 7 days. Inoculate 56.4 μg bacterial cell homogenate group (equivalent to 2×10 8 Plague attenuated EV strain), thalline homogenate+V+F1 vaccine group and negative control group, no lumps were touched at the injection site, and the animals had no reverse hair, diarrhea, or other poisoning phenomena. while inoculating 1×10 7 Three of the animals that received live attenuated plague EV vaccine died, and ...

Embodiment 3

[0059] Example 3 Immunogenicity Detection

[0060] In order to study the immunogenicity of the vaccine of the present application, the experiment was divided into four groups, with 10 female Balb / c mice in each group, respectively:

[0061] 1) Negative control group: normal saline 0.1ml / body

[0062] 2) Cell homogenate+V+F1 vaccine group: cell homogenate 30μg+F1, V each 10μg / 0.1ml / body

[0063] 3) F1+V group (combination of F1 and V antigens): 10 μg / 0.1ml of F1 and V each

[0064] 4) Al-(F1+V) group: Al(OH) 3 +F1, V each 10μg / 0.1ml / piece

[0065] Mice were immunized subcutaneously on the back on day 0 and day 21, respectively. Blood was collected within 1, 2, 3, 4, and 5 weeks after immunization, and serum antibody titers were measured by ELISA. The results of F1 and V antibody titers are shown in Table 1 and Table 2, respectively.

[0066] Table 1 F1 antibody titer (log10)

[0067]

[0068] Note: * Indicates that compared with the cell homogenate+V+F1 group, P<0....

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Abstract

The invention provides a composite plague vaccine and the preparation method thereof. The plague vaccine comprises thallus split products of the attenuated strain of plague bacilli and recombination plague bacilli V antigens, and further comprises plague bacilli F1 antigens, wherein, by collecting the thallus of plague attenuated strain grown on the culture medium, the thallus split products of the plague attenuated strain is obtained through the cracking preparation by high-pressure homogenization. Bacterial cell wall with good adjuvant effect, bacterial protein with antigenic specificity (which includes protective capsular antigen F1) and CpG fragments which are rich in bacterial DNA, with stronger immunological competence, are concentrated in the split products; the expression vector including plague bacilli V antigens or F1 antigens which are constructed by adopting the gene recombination technology is transformed into escherichia coli, and after the inducible expression is performed, recombination plague bacilli V antigens or F1 antigens are obtained through purification. The vaccine is applicable to the general population for immunizing and preventing the plague.

Description

technical field [0001] The invention relates to a plague vaccine used for the immune prevention of human or animal from Yersinia pestis infection, especially suitable for the immune prevention of normal people, and also relates to the preparation method of the vaccine. Specifically, the vaccine contains the cell lysate of the attenuated strain of Yersinia pestis and the Yersinia pestis V antigen, and optionally also includes the Yersinia pestis F1 antigen. Background technique [0002] Plague is a flea-mediated zoonotic disease caused by the bacterium Yersinia pestis (also known as Yersinia pestis). There have been three plague pandemics in the world. In the second epidemic that broke out in Europe in 1348, 30% of the European population died. In past plague pandemics, hundreds of millions of people died of plague. Plague among humans is rare now, but plague among rats is still rampant. The natural foci of plague worldwide have not shrunk. In addition, economic difference...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/02A61P31/04
CPCY02A50/30
Inventor 王国治魏东王秉翔
Owner NAT INST FOR THE CONTROL OF PHARMA & BIOLOGICAL PROD
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